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1939

A Laboratory Manual of Vertebrate Embryology

F.B. Adamstone

Waldo Shumway

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Adamstone, F.B. and Shumway, Waldo, "A Laboratory Manual of Vertebrate Embryology" (1939). Jane
Claire Dirks-Edmunds Documents. Document. Submission 10.
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A LABORATORY MANUAL OF
VERTEBRATE EMBRYOLOGY

I
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 A LABORATORY MANUAL
OF

VERTEBRATE EMBRYOLOGY
ANATOMY OF SELECTED EMBRYOS· OF THE
FROG, CHICK, AND PIG

BY
F. B. ADAMSTONE, Ph.D.
ABSISTANT PROFESSOB OF ZOOLOaY, UNIVERSlT'Y OF ILLINOIS

AND

WALDO SHUMWAY, Ph.D.

PROFESSOB OF ZOOLOGY, UNIVERSITY OF ILLINOIS

NEW YORK

JOHN WILEY & SONS, INC.

LoNDON: CHAPMAN & HAIL, LDmm>
1939
 CoPYIUGBT, 1989, BY

FRANK B. ADAMSTONE
..um
WALDO SHUMWAY

AU RiqhU Reserved
TIM book or any part tli.ereof mtu1t Ml
be reproduced in any form wit1wut
the written permiaNn of the publillh#.

PRINTED IN U.S. A.

PAE&G OP'
BRAUNWORTH 81 CO., fNC.
aun..DERS
OP' BOOK•
8AIDOEPORT* CONN.

CERCLA BINDING U. S. PAT. NO. 1968''7

 FOREWORD

In teaching laboratory classes in vertebrate embryology the instructor is confronted with a number of
difficulties, chief of which is the fact that most students have had no previous experience with the meth-
ods which must be employed. This manual attempts to introduce these methods and to substitute a thor-
ough, careful study of the anatomy of certain developmental stages of the frog, chick, and pig for the
time-honored method of drawing a few individual and supposedly representative sections of these forms.
The outline has been developed over a period of time comprising the past ten years. The results of this
work were first embodied in a manual on the chick and 10-mm. pig published by one of us (F. B. A.)
in 1938. After a year's employment in the laboratory, the original work has been enlarged and a chap-
ter devoted to the anatomy of frog embryos has been added. All drawings have been specially prepared
by us, and those of models depict wax reconstructions made by students in our courses.
The preliminary work on frog eggs and embryos up to the 6-mm. stage and the 18-, 24-, and 33-hour
chicks is carried out by the old method for three reasons, namely: that the embryo is still comparatively
simple, the student is usually unfamiliar with any other method of study, and in addition the old method
gives the student an opportunity to become familiar with the general appearance of the structures which
are most frequently encountered in the more intensive study of serial sections.
At the 6-mm. frog and the 48-hour chick stage the transition is made from the old to the new method.
The student is expected first to draw a number of representative sections in the orthodox manner, but
he must, before this, study carefully the numbered drawings presented iri order to be able to recognize
structures in the section. After this has been done he is required to study the entire series, make a record
of the struct~s identified, and begin the process of learning to recognize them by means of their diag-
nostic features and their relation to each other. The study of the 11-mm. tadpole, the 72-hour chick, and
the 10-mm. pig dispenses entirely with drawings except for those of the total mount and for the informal
sketches made by the student as reference records. Marginal spaces are provided for these records and
sketches. We have prefixed an asterisk to certain structures of which we ask the students in our labo-
ratory to make informal sketches in the marginal spaces opposite. It is also suggested that all un-
named structures in the text figures should be carefully labeled by the student. Drawings and similar
records are not overemphasized, and the success or failure of the student is determined by individual
quizzes.
As here organized, the material for laboratory work is divided into three parts, treating the em-
bryology of the frog, chick, and pig respectively. Since in many courses time does not permit the
intensive study of more than two forms, the part dealing with the frog or pig may be omitted at the
choice of the instructor. In our own laboratory, sections on the 6-mm. and 11-mm. frog embryo are
omitted in order to permit the students to make their own preparations of the 72-hour chick and
10-mm. pig.
The following programs are suggesled:
A. For a course including 6 clock hours in the laboratory, any of the following combinations:
1. Frog (complete) and chick.
2. Frog (omitting 11-mm.), chick, and pig.
3. Frog (omitting 6- and 11-mm.), chick, and pig, including practice in microscopical tech-
nique. This program is used by the authors.
B. For a course including 4 clock hours in the laboratory, any of the following combinations:
4. Chick and pig.
5. Frog (omitting 11-mm.) and chick.
F. B. A.
W.S.
URBANA, ILLINOIS
1939
 CONTENTS

PART I. ANATOMY OF FROG EMBRYOS

PAGE
THE GERM CELLS AND FERTILIZATION 1
CLEAVAGE 2
GERM LAYER FORMATION 3
NEURAL TUBE FORMATION 5
3-MM. EMBRYO 6
6-MM. EMBRYO 11
11-MM. EMBRYO 19

PART II. ANATOMY OF CHICK EMBRYOS

THE GERM CELLS AND FERTILIZATION 31

CLEAVAGE AND GERM LAYER FORMATION 32
18-HOUB EMBRYO 33
24-HOUR EMBRYO 33
33-HOUR EMBRYO 38
48-HOUR EMBRYO 40
72-HOUR EMBRYO 56

PART III. ANATOMY OF THE 10-MM. PIG

EXTERNAL FoRM '\

67
INTERNAL ANATOMY 67

vii
 LABORATORY MANUAL OF VERTEBRATE
EMBRYOLOGY
PART I

ANATOMY OF F ROG EMBR YOS

THE GERM CELLS AND F E RTILIZATI ON
T he Ovum
The frog's egg is spherical with a diameter of 1.7 mm. Its upper or animal hemisphere is black;
the lower or vegetal hemisphere is white. It is surrounded by a very thin envelope, called the vitelline
membrane, which is tightly pressed against the egg. Outside this is a thicker and tougher layer, the
chorion, which is not to be confused with the chorion formed by developing birds and mammals. Out-
side the chorion is the egg jelly, w~.ich consists of an inner and an outer
layer. Thes~ egg envelopes are best observed in living eggs. Nucleus
The nucleus, which lies in the animal hemisphere, can be seen in
sections of ovarian or uterine eggs. This hemisphere is marked by the
dark pigment granules lying just under the periphery. Scattered through
the cytoplasm of the egg are the ovoid yolk granules.
The Spermatozoon
The male germ cell is extremely minute, approximately 0.1 mm. in
length. There is a long cylindrical head containing the nucleus and a
slender tail which is twice the length of the head. a b
FIG. 1. Germ cells of the frog (Rana
F ertilization ( --r I , .. -pipiens). a, Cross-section of ovarian
Under natural conditions the eggs of the frog mature simultaneously egg (approximately 25 X); b, Sper-
in the spring, a condition which may be brought about in the laboratory matozoon (approximately 1000 X).
by injections of pituitary gland into the body cavity.
1

The eggs escape from the follicles of the ovary and are collected in the oviducts. When stimulated
by the embrace (amplexus) of the male, the eggs are discharged from the cloaca, while the sper-
matozoa of the male are simultaneously emitted as a stream of milt. Pituitary-induced eggs may be arti-
ficially fertilized with spermatozoa removed from the sperm ducts of the male.
In living material it is possible to see at the animal pole two polar bodies, one formed before,
and one after, fertilization. One can also recognize a narrow gray crescent lying between the black
animal and white vegetal hemispheres on one side of the egg. This is the side opposite the one on
which the sperm entered the egg. These observations are extremely difficult to make on preserved eggs.
As the eggs enter the water, the egg jelly swells and the outer layer becomes sticky so that all the
eggs from one ovary tend to form a large mass of spawn containing about 2000 eggs.

Laboratory Directions
Examine the demonstrations of eggs and sperm provided.
1 SeeR. Rugh, "Embryonic Material for Laboratory Ex~erimentation," Am. Biol. Teacher,-1:8:184, 1939.
2 ANATOMY OF FROG EMBRYOS

CLEAVAGE
First Cleavage. A furrow appears in the animal hemisphere about
21h hours after fertilization if the eggs are kept at a constant
temperature of 18° Cent. This furrow grows down through the
vegetal hemisphere, thus separating the egg into the first two
blastomeres. In fresh material it can be seen readily that this
furrow usually passes through the gray crescent.
fttw/

Second Cleavage. The s cond cleavage plane also commences at

the animal pole, crossing the first at right angles and dividing the
egg into four blastomeres. Careful inspection of the animal
hemisphere shows that the two furrows do not meet in an exact
cross. Two of the blastomeres are in close contact, forming a
short longitudinal groove known as the polar furrow.

Third Cleavage. This stage is marked by the formation of a lati-

tudinal furrow slightly above the equator of the egg and lying in
the animal hemisphere. It divides the egg into eight blastomeres.
Careful examination shows that this furrow is not an exact circle
but has irregularities where it intersects the ether cleavage planes.
 GERM LAYER FORMATION 3

Fourth Cleavage. Two furrows appear at this stage, beginning in

the animal hemisphere, and grow down through the vegetal hemi-
sphere to produce sixteen blastomeres. These furrows may inter-
( sect each other at the animal pole or may be parallel to each other.
'-I Careful examination of the animal hemisphere should be made.
()'" \
~ Filth Cleavage. Again two furrows are formed, one in the animal
hemisphere and one in the vegetal. The one in the animal hemi-
sphere appears first. On the completion of the lower furrow,
thirty-two blastomeres are formed. Those in the animal hemi-
sphere are smaller and are called micromeres; those of the vegetal
hemisphere are larger and are called macromeres. From this
stage on increasing irregularity of the cleavage planes is apparent.

Blastula. The fifth cleavage stage is followed by the sixth, etc.,

until the end of the cleavage period comes with the formation of
the blastula. In this stage the pigmented area has increased at
the expense of the unpigmented area. The cells in the pigmented
area are so small that they are seen only with difficulty even
when observed with a hand lens. If the blastula is split with a
safety-razor blade through the animal pole, 1 it will be seen that
it contains a large cavity, the blastocoel. The roof of this cavity
(animal hemisphere) i very thin; its floor (vegetal hemisphere)
is very thick. This is because of the concentration of yolk in the
macromeres of the vegetal hemisphere.
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Laboratory Directions
Examine the stages described above and make outline sketches (about one inch in diameter) of each
in the space provided opposite each of the paragraphs above.
GERM LAYER FORMATION
After the formulation of the blastula the embryo is converted
into a gastrula with several germ layers. Externally the process of
germ-layer formation or gastrulation is marked by the progressive
formation of the blastopore, conveniently divided into three stages:
(I) the appearance of the dorsal lip (24 hours at 18° Cent.); (2)
the development of the lateral lips from the dorsal lip; and (3) the
formation of the ventral lip by the union of the lateral lips.
The study of sections shows that at the dorsal lip, surface cells
roll inward to form the roof of a slitlike cavity, the gastrocoel. As
the dorsal lip advances over the surface of the vegetal hemisphere,
it also elongates to form the lateral lips, where a similar in-rolling
occurs, thus adding to the side walls of the gastrocoel. In its ad-
vance the dorsal lip moves almost completely over the vegetal hemi-
sphere so that when the lateral lips unite to form the ventral lip,
the completed lip of the blastopore forms a small circle enclosing
the only white area left on the surface of ~he egg. This white area
1 This may be done by demonstration.
ANATOMY OF FROG EMBRYOS

which protrudes slightly above the level of the blastopore is the

yolk plug.
The partition between the newly formed gastrocoel and the old
blastocoel is either ruptured or pushed back until the blastocoel is
obliterated, and the gastrocoel is the only cavity persisting. Since
the heavy mass of yolk-laden macromeres is now on one side of the
gastrula, the embryo rotates until the weight is at the bottom, which
brings the blastopore to the posterior end of the embryo (Fig. 2).

f An An

CD v
...

D
...

-D

~ !
I I
Veg. Veg.
D

v
-D
v- An~

Ji. .!!
~
Fm. 2. Germ layer formation in the frog. Diagrammatic. a, dorsal lip stage; b,
lateral lip stage; c, ventral lip stage; d, yolk plug stage after rotation. The smaller
figures to the upper left are surface views; the larger ones to the right are sagittal
sections. An, animal pole; veg., vegetal pole; D, dorsal; V, ventral. The small
arrows indicate the direction of cell movements. The pres'UmptiVe germ layers are
designated after the data of Vogt and Pasteels; epidermal ectoderm, white; neural
plate ectoderm, cross lines; notochord and mesoderm, stipple; endoderm, small circles.

In the frog's egg the layer of cells left on the surface after gas-
trulation is complete is the ectoderm. The layer of cells turned in
at the lips of the blastopore is mes-endoderm. Almost ·as rapidly as
it turns inward, however, it separates into an inner layer which is
the endoderm, lining the roof and sides of the gastrocoel, and an
outer layer between the ectoderm and endoderm which is the
chordamesoderm. The median axial strip of this middle layer is
the primordium of the notochord; the rest becomes the mesoderm
which soon grows out between tbe ectoderm and endoderm in other
regions of the embryo. In an exactly sagittal section it can be seen
that the notochord forms part of the roof of the gastrocoel (Fig. 2d). .
During the formation of the -neural tube the endoderm grows be-
neath the notochord from either side (Fig. 3);a
1
I
 FORMATION OF THE NEURAL TUBE 5

Laboratory Directions
Examine total preparations of the dorsal-lip, lateral-lip, and ventral-lip (yolk-plug) stages of gas-
trulation. Make outline sketches in the space provided as in the last exercise. Prepare a half gastrula 1
by splitting a yolk-plug stage from top to bottom, bisecting the yolk plug. Observe the gastrocoel and
its lining of small white cells, the endoderm. Sketch and label. Examine demonstration slides which
snow the position of all three germ layers.

FORMATION OF THE NEURAL TUBE

(New:al-Stages) /:2
Neural Plate Stage. Soon after gastrulation is completed (56
hours at 18° Cent.) it will be seen that the blastopore is flattened
from side to side to form a slit. Meantime the dorsal surface of
the embryo is flattened to make a plate elevated slightly around
its margin. This flattened area, called the neural plate, is the
primordium of the nervous system.

Neural Fold Stage. The embryo is now changing its shape, elon-
gating along the anteroposterior (cephalocaudal) axis. Alongi-
tudinal depression appears in the middle of the neural plate
forming the•neural groove. The margins of the plate are consid-
erably elevated to form the neural folds. On the dorsal side at
the anterior end of the embryo on either side of the neural folds
is a crescentic elevated area known as the sense plate. Just pos-
terior -to each sense plate and lying alongside the neural folds is
the gill plate. The blastopore has become a very small, incon-
spicuous slit which is now being enclosed between the neural
folds at the posterior end.

Neural Tube Stage. The neural folds have now grown together
to form a neural tube which is elevated above the dorsal surface
of the embryo. This whole process by which the neural plate
sinks in to form the neural groove and is eventually cut off as
the neural tube is an example of a common embryological phe-
nomenon known as invagination. From the dorsal aspect the
embryo is pear shaped and is marked on each side by short trans-
verse grooves representing the anterior and posterior margins of
the gill plates. These grooves are the primordia of the first and
fifth vise_ ral clefts. At the anterior end two swellings may be
seen on the ventral surface. These are the primordia of the
mucous glands. Between them is a depression, the beginning of
the stomodeum. At the posterior end, the blastopore is now
. covered by the fused neural folds, forming a neurenteric canal,
and at the ventral end of the strip resulting from this fusion a
depression indicates the beginning of the proctodeum. In side
view transverse depressions indicate the presence of six somites.
The living embryo is now approximately 2~ mm. in length. The
epidermis has developed cilia by means of which the embryo ro-
tates freely inside the egg jelly.
1 Thls may be shown by demonstration.
6 ANATOMY OF FROG EMBRYOS

FIG. 3. Neural tube formation in the frog (Rana pipiens). a, transverse section
through neural plate; b, transverse section through neural folds and groove; c,
transverse section through neural tube. All at 50 X. Germ layers represented as in
Fig. 2.
Laboratory Directions
Examine the three stages in neural-fold formation described above and sketch each from the dorsal
aspect in the space provided. Examine demonstrations of transverse sectio..ns through each stage.

3-MM. EMBRYO
(Tail Bud Stage)

The embryo of 3 mm. body length is chosen somewhat arbitrarily as an example of an early embryo.
It is about three days old (74 hours at 18° Cent.). Externally it is marked by the possession of a dis-
tinct tail bud. This has grown backward from the posterior end of the fused neural folds. The gill plate
now shows four slight visceral grooves known as the hyoid, first, second, and fourth branchial grooves, re-
spectively. The third branchial groove appears at a later stage. The mucous glands have enlarged con-
siderably. In ventral view, a groove extending from the posterior end forward in each gives them the ap-
pearance of an elongated U. Between them the stomodewn may be identified. At the posterior end of
the body, a short distance ventral and anterior to the base of the tail bud, is the proctodeum, which now
opens into the hind-gut. In lateral view, a swelling in the sense plate region indicates the outgrowth of
the optic vesicle. Posterior to the gill plate and at a level approximating the base of the neural folds a
series of slight transverse grooves marks the presence of twelve or thirteen somites. Ventral to these
and just behind the gill plate is a swelling caused by the developing nephrotomal band. (These details
may be seen better in a cleared and stained specimen.
 3-MM. EMBRYO 7

TABLE 1
PHARYNGEAL DERIVA.TIVES AND ASSOCIATED STRUCTURES

Visceral Visceral Visceral Visceral

(pharyngeal) (branohial) (pharyngeal) Aortic arches (branchial)
pouches arches clefts grooves
Endodermal Ectodermal

-- I
Mandibular
-- I --
I -- I -- I
Hyomandibular Hyomandibular Hyomandibular

-- II
Hyoid
-- II --
II -- II -- II

-- III •
-- III --
... 1st branchial Carotid

III -- III -- III

-- IV -- IV --
2nd branchial Systemic

IV -- IV -- IV

-- v -- v --
3rd branchial

v -- v -- v

-- VI -- VI --
4th branchial Pulmonary

VI -- VI
Lacking in frog
-- VI
Vestigial in frog Lacking in frog

The confusion arising in the minds of students who attempt to do collateral reading concerning the arches and pouches may be avoided by the
study of this table. It will be apfarent that the dilllcnlty centers about the indiscriminate UH of the word "brsnchial" in respect to the arches.
Some authors use the word for al the arche11, others restrict its use to the (liU-b6M"in11 arches as we have done here. We have, however, given the
alternative usage in pareu.thesea, aa we have no desire to appear to force our solution on our colleagues. See also F1g11. 10 and 81.
8 ANATOMY OF FROG EMBRYOS

Laboratory Directions
Make an outline sketch of the 3-mm. embryo from the left side in the space provided below. Label
all structures you have identified.

The 3-mm. Embryo.

SERIAL SECTIONS

The internal anatomy of embryos from this stage onward is studied much better from embryos
which have been cut into a continuous series of sections or slices and mounted in order on slides for mi-

Hypochord-, Notochord Somite Neural tube

~---'....,,,.
..················"....,
' ', ', '------------------.
'',, ''
········ "(' ... ··~· .... '
I ;"":··

.........z.·"
.. ,/
I •.... ·
/,!,.
··............. .
I
/
I
/
Optie Muoous Mid-Igut Epidermis
'
vesicle gland
FIG. 4. Block diagram of 3·mm. frog embryo (Rana pipierui), to show position of
transverse section through liver primordium in relation to the whole embryo repre-
sented as a. transparent body.

croscopic observation. Figure 4 indicates the relationship of a transverse section to the embryo as a
whole. In order to read a series of sections you should examine each section, identifying the structures
you observe.
 3-MM. EMBRYO 9

ECTODERMAL DERIVATIVES

The epidermis is ciliated except on the ventral side, but the cilia themselves are too small to be seen
with your microscope. It will be noticed with high power that the ectoderm is divided into two layers,
the epidermal layer and the nervous layer. The stomodeum, between the two mucous glands, has not
yet joined the fore-gut. From the stomodeum a solid hypophysis grows toward the forebrain. Notice
with high power the structure of the mucous glands. The cells in these glands are more specialized at this
time than any others of the body. The proctodeum has opened into the hind-gut, thus forming the
cloaca. The neurenteric canal has now been closed owing to the growth of the tail bud.
The neural tube is now completely cut off from the exterior. The anterior end or prosencephalon is
bent slightly around the anterior end of the notochord, owing to the cranial flexure. The prosencephalon
extends backward on its ventral side to the posterior level of the optic vesicle. On the dorsal side it ex-
tends backward to the anterior end of a thickening in the roof of the brain. The mesencephalon extends
backward from these boundaries to the anterior level of the notochord. The rhombencephalon begins
at this point. The boundaries of the mesencephalon and rhombencephalon are not easily distinguished
in transverse sections. These and the epiphysis are distinguished more easily in sagittal section. The
spinal cord has grown into the tail bud.
From the ventral halves of the lateral walls of the prosencephalon, the optic vesicles protrude out-
ward, so producing a bulge on either side of the embryo. Farther back, in the region of the rhomben-
cephalon, the nervous layer of the ectoderm is greatly thickened and has invaginated on each side to
form an otic (auditory) pit. Notice that the epidenmal layer of the ectoderm is not invaginated. The
nose at this time is in the form of thickenings of the nervous ectoderm, the olfactory (nasal) placodes, in
sections anterior to, or at, the level of the prosencephalon.

ENDODERMAL DERIVATIVES

The gastrocoel is now lengthened and dilated in the region anterior to the yolk mass to form a fore-
gut. A depression in the floor of the fore-gut extends towards the stomodeum but the two are still sepa-
rated. Posteriorly a fingerlike evagination of the fore-gut extends into the yolk mass as the primordium
of the liver. Frontal sections show the beginnings of the visceral pouches I, II, and III growing out from
each side of the fore-gut towards the corresponding visceral grooves already noticed in the ectoderm.
The mid-gut is formed from the gastrocoel overlying the yolk mass. It has a very thick floor
and relatively thin roof. Above it lies a thin rod much like the notochord but smaller. It is called the
hypochord and is formed from the endodermal roof of the mid-gut, has no known function, and disap-
pears early.
The hind-gut, posterior to the yolk mass, has united with the proctodeum to form the cloaca, open-
ing to the exterior by the small vent al:;eady noticed.

MESODERMAL DERIVATIVES

The notochord extends forward to the base of the mesencephalon where it turns down very slightly.
At the posterior end it joins a mass of relatively undifferentiated tissue which forms the core of the
tail bud.
On either side of the notochord is the axial mesoderm from which twelve or thirteen somites have
been formed. The axial mesoderm terminates in the core of the tail bud mentioned in the last paragraph.
The intermediate mesoderm was partially divided into nephrotomes at an earlier stage. These have
now fused together to form a nephrotomal band. This is best seen in sections cut through the region
of the liver. The lateral mesoderm is divided into an outer thinner somatic layer and an inner thicker
splanchnic layer. No coelom is formed between them as yet except beneath the fore-gut where it is to
be seen in sections cut in the region of the otic pit. Here the sp~e represents the_ beginning of the peri-
cardia! cavity.
10 ANATOMY OF FROG EMBRYOS

Laboratory Directions
Study the set of sections provided you and make sketches of sections through: ( 1) the optic vesicles,
(2) the otic pits, (3) the liver, and ( 4) the cloaca.

.
....

Section through optic vesicles. Section through otic pits.

Section through liver. Section through cloaca.

Transverse Sections of 6-mm. Frog.

 6-MM. EMBRYO 11

6-MM. EMBRYO
(Hatching Stage)

The embryo of Rana pipiens hatches spontaneously when it is 110 hours old (at 18° Cent.) at a
length of 6 mm. Muscular movements may be initiated in this species by stimulation when the em-
bryo is between 3% and 4 mm. in length, and when it is a little more than 5 mm. long the heart starts
beating. The stage now to be described is marked exactly by the beginning of circulation in the capillary
loops of the first external gill.
EXTERN AL FORM

The head is marked off from the trunk by a notch behind the mucous glands which have ap-
proached each other at their posterior ends. The stomodeum, now a deeper pit, is anterior and slightly
dorsal to the anterior ends of the mucous glands but has not yet opened into the fore-gut. On either
side of the stomodeum and somewhat dorsal to it is a nasal pit. On the sides of the head optic bulges are
still prominent. All five visceral groves (hyomandibular and branchials 1-4) have appeared but have
not opened into the corresponding visceral pouches. External gills are developing on the first and
second branchial arches, and circulation can be demonstrated in the first of these.
Transverse intersomitic grooves appear on the trunk, and beneath them a slight ridge marks the
region of the pronephric duct. On the ventral side the cloaca! opening is to be seen just anterior to
the base of the tail.
The tail is now one-half of the body length. !ntersomitic grooves continue into the tail. A prom-
inent tail fin extends completely around the tail from the vent on the ventral side to the dorsal junction
with the truiik.

Laboratory Directions
Make a drawing of the 6-mm. embryo from the left side.

The 6-mm. Embryo from the Left Side.

12 ANATOMY OF FROG EMBRYOS

REPRESENTATIVE SECTIONS OF THE 6-MM. EMBRYO

The sections described and illustrated are selected from a series of 275 sections cut transversely
to the long axis of the embryo at a thickness of 20 microns (0.02 mm.). They are reproduced from
preparations made to compensate for the reversal produced by the compound microscope. Right on
the illustration is right on the embryo. The student is cautioned that because of slight variations in
the developmental tempo of embryos and variations in the plane of section he will probably never
encounter sections exactly like those illustrated.
A. Section through the Epiphysis and Nasal Pit (Section 10, Fig. 5)
A slight lateral curvature of the head results in this section's showing a slight asymmetry, the
plane of section passing through the left side at a region more anterior than that on the right. The
nasal pit ( 1), therefore, is cut on the left through its opening to the exterior, and on the right through
its posterior side wall. The groove on the ventral side is the beginning of the stomodeum (2). The
brain is cut through its most anterior division, the telencephalon (3), at the level of the primordium
4

Fm. 5. Rana pipiens, 6-mm. em- Fm. 6. Rana pipiews, 6-mm. embryo.
bryo. Transverse section through Transverse section through optic cup.
nasal pit. 50 X. 50X.

of the pineal gland, or epiphysis ( 4). An examination of the epidermis (5) with the high power will
reveal a few larger rounded cells, projecting slightly from the surface, which bear cilia. The space
between these structures (all of ectodermal origin) is filled with a loosely connected tissue, the mesen-
chyme (6), partly of mesodermal derivation and partly from the ectoderm of the neural crest. A few
small cavities represent the neural capillary plexus (7) from which the blood vessels of the head are
forming at this time.
B. Section through Optic Cup and Lens Vesicle (Section 22, Fig. 6)
There is still a slight asymmetry so that the section passes through the lens vesicle (1) on the
left and behind it on the right. The vesicle has just detached itself from the epidermis and has a small '
central cavity. The optic cup has two layers, an outer thicker sensory layer (2a) and an inner, very
thin, heavily pigmented one (2b). The neural tube is partially divided by a constriction into the ven-
tral diencephalon (3) and the more dorsal mesencephalon (4). Beneath the floor of the diencephalon
lies a fiat thick plate of cells, the hypophysis (Rathke's pocket), (5). The epidermis (6), with its cili-
ated cells, covers the surface of the embryo. On the ventral side are two large mucous glands (7), the
one on the right cut through its cuplike groove.
•
I

'
 6-MM. EMBRYO 13
In the ventral half of the section is the large cavity of the thin-walled pharynx (8) and beneath
it a solid laterally compressed rod, the thyroid gland (9).
Between these structures lies the mesenchyme (10) containing a few blood vessels. Those lo-
cated between the eye and brain are parts of the ophthalmic vein (12) which lead to the anterior
cardinals (see later). Those between the brain and the pharynx are part of the cerebral arteries (11)
which represent anterior extensions from the carotids (see later).

C. Section through Otic Vesicles, Heart, and External Gills (Section 38, Fig. 7)
At this level the curvature which resulted in the asymmetry of the two preceding sections has now
been passed. As this section passes through the two otic vesicles (1), the portion of the brain between
them must be referred to the myelencephalon (3) with its thin-walled roof. A slight projection on the
inner dorsal angle of each otic vesicle is the beginning of its endolymphatic duct (2).
Beneath the brain is the conspicuous notochord ( 4) with
large vesicular cells. Beneath this in turn lies the widely ex- 8
I
panded pharynx ( 5). The lateral thickenings of its walls repre-
sent the fourth visceral pouch (6). Ventral to this in the cavity
of a great bulging projection lies the heart, represented by the
dorsal atrium (8) and ventral ventricle (7). It is contained in
the most anterior part of the coelom, the pericardia! cavity (9).
Note that this cavity is lined on the outside by the somatic layer
of the mesoderm ( 10) and on the inner side by Ute splanchnic
layer of the mesoderm ( 11), which gives rise to the myocardium
and visceral -pericardium. The endocardium (12) separated at
an earlier stage. --15
From either side of the embryo projects the first external
gill (13) borne on the third visceral arch. This gill is now branched
and has several capillary loops (14). Circulation in this gill can
be seen in the living embryo. The capillary loops receive blood
from the first afferent branchial artery (15), which is beneath the
pharynx. The capillary loops of the gill discharge their blood to
the first efferent branchial artery (16), which is dorsal to the side 9
of the pharynx. These arteries represent the third aortic arch Frn. 7. Rana pipiens, 6-mm. embryo. Trans-
verse section through otic vesicle, gills and
of vertebrates. Between the otic vesicle and the brain may be heart. 50 X.
seen portions of the internal carotid arteries (18) and at the
ventro lateral margins of the otic vesicles the anterior cardinal veins (17) appear. The mesenchyme
and epidermis are readily identified.

D. Section through Pronephros and Liver (Section 55, Fig. 8)

This section cuts through the region where the liver (1) joins the intestine (2). The brain is cut
in the region of the myelencephalon (7) as shown by the thin roof. Lying beneath it is the notochord
(9) and just below this is a smaller rod, the hypochord (10), formed from the roof of the gut but now
separated from it by the dorsal aortae (6) which are here uniting together.
On either side of the notochord and stretching up alongside the myelencephalon is a myotome (8),
with which the dermatome is still associated. The intermediate mesoderm or nephrotomal band is
greatly enlarged, and in it have appeared the three pronephric tubules (5). One of these is cut to
show the nephrostome (3) or opening into the coelom (13). This cavity is lined on the outside by
the somatic layer of the mesoderm (14) and on the inside by the splanchnic layer (15). Note the
slight bulges at the inner dorsal .angles of the coelom caused by the outward growth of each glomus
( 4) from the dorsal aorta. Above and to the outer side of each glomus, the large anterior cardinal
veins (11) appear. The posterior cardinal veins (12) are seen beneath the pronephric tubules. On
14 ANATOMY OF FROG EMBRYOS

the ventral side of the liver, enclosed by the splanchnic layer of the mesoderm, are the large vitelline
veins (16). The epidermis (17) presents nothing different from other sections.

E. Section through Proctodeum (Section 159, Fig. 9)

If it were not for the presence of the proctodeum (1) this would be a typical section through
the tail region. Note the tail fin (2) above and below. In the mid-region are seen the spinal cord

---2

I ,
I I
I /
v F1a. 9. Rana pipiens,
16 6-mm. embryo.
Fm. 8. Rana pipiens, 6-mm. embryo. Transverse section
through tail at level
Transverse section through liver and of cloaca! opening.
pronephros. 50 X • 50X.

(3), notochord (4), and hypochord (5). On either side is one of the tail myotomes (6). Beneath
the hypochord is the small caudal artery (7), and some little distance beneath this the caudal vein (8).
The tail fin is composed chiefly of mesenchyme (9) and epidermis (10) with many ciliated cells.

THE STUDY OF SERIAL SECTIONS

After studying the representative sections just described the student should be ready to survey
the anatomy of an embryo from a complete set of serial sections. Such a set consists of a complete set
of slices across the long axis of the embryo arranged in order from the tip of the head to the end of
the tail. Everything which could be found by dissection, if the embryo were larger, can be identified in
some part of the series. Once identified the structure can be traced forward and backward until a
complete mental picture of the organ and its relationship to others is formed. It is advisable before
proceeding further to identify the plane of sectioning by laying a ruler across the lateral view of the
whole embryo you have drawn earlier and tracing lines at the levels of the representative sections just
studied.
As you succeed in identifying the structures mentioned in the following paragraphs, make a record
of each, by writing in the blank spaces provided: ( 1) the number of the slide in the set you use;· (2)
the number of the row of sections in which you first identify the structure (counting from top to bot-
tom); and (3) the number of the section in the row (counting from left to right). A certain number
of the structures are starred. It is recommended that you make in the margin of this manual opposite
the starred structure a simple line sketch to show its appearance in the section you have identified.
 6-MM. EMBRYO 15

ECTODERMAL DERIVATIVES

I. EPIDERMAL STRUCTURES
a. Epidermis. A single layer of ectoderm cells forming the outer
covering of the body, many of them ciliated.
b. Stomodeum. A pit on the ventral anterior surface of the head
separated by a plate ( ectoderm on the outside, endoderm on the
inside) from the fore-gut.
c. Proctodeum. A pit on the ventral surface of the trunk where
it joins the tail, opening directly into the hind-gut to form the
cloaca.
d. Mucous glands. Prominences ·on the ventral side of the head
on either side of the stomodeum and behind it. Observe deep
groove in each.

II. BRAIN AND SPINAL CORD. Starting at anterior end identify:

a. Telencephalon. First portion of the brain to appear in the series.
b. Diencephalon. Commences at the point where the epiphysis
joins the brain. From its sides the optic stalks emerge. Beneath,
a backward groove finally separates from the diencephalon to
form the infundibulum.
1. Epiphysis. Extends forward from its point of origin. Now
has lost its cavity.
* 2. lnfundibulum. Find a section where the brain appears in
two parts. The ventral portion is closely applied to the
hypophysis (see below).
3. Hypophysis (Rathke's pouch), a derivative from the anterior
end of the stomodeum. It will unite with the infundibulum
to form the pituitary gland.
c. M esencephalon. With a thick roof. Lies between optic cups and
above the infundibulum.
d. M etencephalon. A thin roof. Lies between otic vesicles.
e. Myelencephalon. A thin roof. Hardly to be distingui3hed from
metencephalon. Continues backward to spinal cord.
f. Spinal cord. Thick walls, thinner roof and floor. Continues back
into tail almost to the end.

III. SENSE ORGANS OF THE HEAD

a. Nasal (olfactory pit). Most anterior sense organ, on ventral
side of head, marked by thickness of epithelium and opening to
the exterior only.
b. 1. Optic cup. Consist of pigment layer and thick sensory layer,
joined to diencephalon by optic stalk.
2. Lens vesicle. With central cavity, located in mouth of optic
cup and separated from epidermis.
c. Otic vesicle. Located alongside myelencephalon. A thick-walled
hollow vesicle with an endolymphatic duct just forming.
rv. CRANIAL NERVES. At this stage there are four masses of tissue resembling the tissue of the walls of
the brain in the mesenchyme on either side of the metencephalon and myelencephalon. These are
ganglia which will supply sensory neurones to certain cranial nerves. •
16 ANATOMY OF FROG EMBRYOS

a. Anterior to the otic vesicle and proceeding forward

* 1. Facial-acoustic complex. Immediately in front of the otic
vesicle. Supplies afferent fibers to cranial nerves VII and
VIII.
* 2. Trigeminal ganglion. Very large. Supplies afferent fibers to
cranial nerve V.
b. Posterior to the otic vesicle and proceeding backward
* 1. Glossopharyngeal ganglion. Just posterior to the otic vesicle
and ventral to it. Supplies afferent fibers to cranial nerve IX.
* 2. Vagus ganglion. Some distance farther back and a little
dorsal to the last. Supplies afferent fibers to cranial nerve X.

ENDODERMAL DERIVATIVES

a. Mouth. The region of the fore-gut just behind the plate sep-
arating it from the stomodeum will form the oral cavity of the
mouth. This continues as pharynx.
b. Pharynx. This region with its visceral (pharyngeal) pouches
is best seen in ~rontal section (Fig. 10). Lay your ruler across

- Nasal pit
VISCERAL
ARCHF.S

m
m
II
'ol·
Frn. JO. Rana pipiens, 6-mm. embryo. Frontal section through head at level of vis--
ceral pouches and arches. 50 X .

this figure and draw a line to show the position of the transverse
section shown in Fig. 7.
c. Thyroid gland. Originates from the floor of the pharynx as a
solid rod from the level of the second visceral pouch to the fifth.
d. Liver. The liver diverticulum noted in the 3-mm. embryo has
grown forward towards the heart. The posterior ventral portion
shown in Fig. 8 will become the gall bladder.
e. Intestine. From the liver backward the mid-gut becomes the
intestine, with a small cavity and an exceedingly thick floor
which contains the yolk.

'
 6-MM. EMBRYO 17
f. Cloaca. Starting with the proctodeum trace the cloaca forward
until it unites with two small longitudinal tubes, the pronephric
ducts (see later).

g. H ypochord. This is a small rod of endoderm separating from

the roof of the gut and lying beneath the notochord. It extends
back into the tail.

MESODERMAL DERIVATIVES

1. NOTOCHORD. Cylindrical rod with characteristic vesicular cells. Lying beneath brain and spinal
cord. Commencing behind infundibulum and extending well back into the tail.
n. MESENCHYME. Loose reticular embryonic connective tissue filling space between organs.
III. DERIVATIVES OF THE SOMITES. The somites extend from behind the otic vesicle well into the tail.
There are thirteen somites in the trunk and approximately twenty more in the tail. The inner
ventral portion of each somite (sclerotome) is now broken down into a mesenchymal tissue invest-
ing the notochord and neural tube from which the axial skeleton will be formed. The dermatome
or outer layer (from which the dermis of the skin ~ill develop) is still associated with the muscle-
forming remainder of the somite or myotome. Jn these myotomes muscle fibers are developing.
IV. DERIVATIVES OF THE INTERMEDIATE MESODERM. In the frog this region forms the nephrotomal band
from whic'k the kidney and gonads develop.
a. Pronephric tubules. Three on each side. Well-defined wall of
thick cells. Pigment concentrated at the cell margin nearest
the cavity. Communicates with the coelom by funnel (nephro-
stome). It is not ciliated at this time.
b. Pronephric duct. Originates from the last pronephric tube and
continues in the ventral portion of the nephrotomal band. Trace
the pronephric duct to its junction with the cloaca.

V. DERIVATIVES OF THE LATERAL MESODERM

a. Somatic mesoderm. Closely applied to the ectoderm in the trunk.

b. Splanchnic mesoderm. Closely applied to the endoderm in the
trunk region.
c. Coelom. The cavity located between the somatic and splanchnic
layers of mesoderm. The anterior portion of the coelom known
as the pericardial cavity surrounds the heart. This coelom is not
to be found in the head or tail.
d. M esenteries. The right and left splanchnic and somatic layers
of the mesoderm unite above the intestine and heart. These
regions constitute the dorsal mesentery and dorsal mesocardium
respectively.
v1. CIRCULATORY SYSTEM. The blood vessels and lymphatics of the frog embryo originate in a system
or network of capillaries (plexus) associated with the organ systems and derived from splanchnic
mesoderm in the trunk region or from mesenchyme in the head or tail. They are best demonstrated
by injection 1 preparations.
1
H. M. Knower, "A Resurvey of the Developments of Lymphatics and Associated Blood Vesiels in Anuran Amphibia
by the Method of Injection," Wistar Institute, 1939.
18 ANATOMY OF FROG EMBRYOS

a. Heart. Locate the heart, at the level of the external gills, pro-
ceeding backward until it divides into two large vessels entering
from the liver.
* 1. Sinus venosus. The point where the two vessels converge,
forming a stubby inverted Y.
2. Atrium. The sinus continues forward and dorsal, bending to
the right, to enter the thin-walled atrium.
3. Ventricle. The heart in more anterior sections takes on the
appearance of a reversed C. The lower limb bending from
the atrium to the left side is the ventricle.
* 4. Bulbus arteriosus. Continuing forward, the lower left por-
tion of the reversed C becomes detached from the atrium and
is now known as the bulbus.

b. Arteries
1. Of the head
* Ventral aorta ( truncus arteriosus). Following the bulbus
towards the head, it immediately assumes the form of a
letter T. The base represents the bulbus, the shaft the
short ventral aorta, and the right and left arms at the
top are the branches of the truncus which later subdivide
to form the afferent branchial arteries.
Afferent branchial arteries. Three on each side, located in
the first, second, and third branchial arches (visceral
arches III, IV, and V). Those of the fourth branchial
arch have not yet appeared.
Efferent branchial arteries. Also three on each side, con-
nected by capillary loops with the afferent branchials. 1
* Radices aortae. The efferent branchial arteries on each
side unite to form an aortic root.
Dorsal aorta. Formed by the union of the right and left
radices aortae in the region of the pronephros.
Internal carotid arteries. These are anterior prolongations
of the radices aortae.
2. Of the trunk
* Glomus. This is a short side projection from the point
where the radices aortae are fusing, bulging out into the
coelom opposite each pronephros.
3. Of the tail
Caudal artery. The direct continuation of the dorsal aorta.
c. Veins
1. Splanchnic
V itelline ( omphalomesenteric) veins. These arise on the
ventral side of the thick-walled gut and pass forward
around the liver to join the heart at the sinus venosus.
2. Somatic (cardinal system)
Anterior cardinal veins. These collect the blood from nu-
1 The afferent and efferent branchial arteries are the representatives of the aortic arches III,

oped in vertebrate embryos.

IY, and V, generally devel- II
 11-MM. EMBRYO 19

merous small tributaries in the head and convey it back-

ward as far as the pronephros, where they turn and bend
sharply downward to course along the anterior surface
of that organ.
Posterior cardinal veins. These vessels course forward from
the region of the cloaca ventral and lateral to the dorsal
aorta, passing along the ventral surface of the pronephros
until they meet and join the anterior cardinal veins.
Common cardinal veins (ducts of Cuvier). These vessels,
formed by the union of the anterior and posterior cardi-
nals on each side, course downward and enter the sinus
venosus at points just dorsal to and lateral of the vitelline
veins.
Caudal vein. This courses through the tail just ventral to
the caudal artery. It divides into two branches which
unite with the posterior cardinal veins on meeting the
cloaca. Further details of the smaller arteries, veins, and
lymphatics are omitted. The more advanced student is
referred to the textbook ·and its references.

11-MM. EMBRYO
(Larval Stage)

The embryo of Rana pipiens attains the length of 11-mm. after 284 hours at a temperature of
18° Cent. At this stage it has assumed the familiar tadpole shape with head and trunk forming an
ovoid compressed dorsoventrally whereas the tail~ compressed laterally, is twice the length of the body.
The mouth is open and surrounded by horny raspers or oral combs. The mucous glands have degen-
erated but persist as two small vestiges. The large eyes protrude slightly. The openings of the nose,
external nares, are located anterior to the eyes. ·The external gills are covered by folds developed from
the hyoid arches (visceral arch II). This fold is fused with the ectoderm of the body on the right side,
but on the left an opening, the spiracle, or opercular aperture, persists. At the base of the tail is the
vent or cloaca! aperture. On either side and dorsal to the vent are the buds of the posterior limbs.

Laboratory Directions
Make a drawing of the U-mm. embryo from the left side (page 20).

DIRECTIONS FOR STUDY OF TRANSVERSE SERIAL SECTIONS

The 11-mm. tadpole is a larval stage in the development of the frog. It is a free-swimming animal
carrying on all the functions of animal life except that of reproduction. It is to be expected, therefore,
that its internal anatomy will be more complex than that of the embryos studied heretofore. Accord-
ingly drawings of certain organ systems are provided, based on wax reconstruction~. The student is ex-
pected to trace the organ systems through the series of transverse sections, noting in the marginal
blanks the number of the slide, row, and section in which each particular structure is well represented.
It is suggested that the study be commenced by examining a typical section of the tail and comparing
it in detail with the tail of the 6-mm embryo. It will usually be easier to commence at the region
of the cloaca and trace all structures forward. •
20 ANATOMY OF FROG EMBRYOS

The 11-mm. Embryo from the Left Side.

ECTODERMAL DERIVATIVES

I. EPIDERMIS. No longer ciliated except on the tail.

II. SPIN AL CORD AND BRAIN
a. Spinal cord. Extends forward until the roof thins.
b. M yelencephalon. The cavity widens in more anterior sections.
The thin roof is the primordium of the choroid plexus of the
·fourth ventricle.
c. M etencephalon. Distinguished with difficulty from the myel-
encephalon. Located between the otic vesicles. At the anterior
end, the roof thickens suddenly to become the primordium of
the cerebellum.
* d. Mesencephalon. Just anterior to the thickened primordium of
the cerebellum, two swellings form the optic lobes (corpora bi-
gemina) of the mesencephalon. The roof of the mesencephalon
expands widely anterior to the optic lobes. Its cavity is the
mesocoel, iter, or aqueduct.
e. Diencephalon. Roof narrower than that of mesencephalon.
Cavity called diacoel. Anterior boundary marked by epiphysis.
1. Epiphysis. Projects dorsally and anteriorly from roof of
diencephalon. -
 11-MM. EMBRYO 21

2. Infundibulum. Broad evagination from floor of mesenceph-

alon. Extends posteriorly beneath mesencephalon as far as
anterior end of notochord.
* 3. Optic chiasma. A swelling on each side of the floor of dien-
cephalon anterior to infundibulum. Marks point of entrance
of optic nerve.
4. Optic recess. Small evagination from floor of diencephalon.
Marks anterior boundary.
f. Telencephalon. Its posterior boundary is the optic recess. Roof
thin. From roof a folded vascular membrane called the anterior
choroid plexus hangs down into cavity known as telocoel. Telo-
coel and diacoel (see above) make up third ventricle.
* 1. Cerebral hemispheres. Formed by dorsal anteroposterior con-
striction from roof of telencephalon. Cavity of each called
lateral ventricle.

Nasal tube-

Diencephalon -

Fxo. 11. Brain and sense organs of 11-mm. Rana pipiens embryo. From a wax plate
:teconstruction. Dorsal view. (NOTE: Cranial nerve X, closely associated with IX,
" does not appear from this view.)

III. CRANIAL NERVES

a. Associated with nose
(I) Ophthalmic (olfactory). From ventral side of each cerebral
hemisphere at its anterior end and passing directly to the mesial
side of each nasal tube.
b. Associated with eye and listed in order from front to rear
* (II) Optic. From optic chiasma of diencephalon lateral and
posterior to each eyeball. Can be seen passing through retina.
(III) Oculomotor. Very small. Emerges from floor of dienceph-
alon on each side, just dorsal to infundibulum.
(IV) Trochlear. Very small. Emerges from dorsolateral angle
of metencephalon, just posterior to optic vesicles.
22 ANATOMY OF FROG EMBRYOS

(VI) Abducens. Very small. Emerges from floor of myelen-

cephalon, just anterior and ventral to root of l:a.rge trigeminal
nerve (see below).

NoTE: These three nerves supply the muscles of the eyeball. They are very small and require the highest power lenses
of the student microscope.

c. Associated with ear and listed in order from front to rear

1. Just anterior to ear.
(V) Trigeminal. Large nerve with ganglion. Arises from
ventrolateral angle of myelencephalon. Sends branches to
maxillary and mandibular processes of first visceral arch.
2. Between ear and myelencephalon. Closely associated with
each other.
(VII) Facial. Arises from side of myelencephalon. Sends
branches towards trigeminal and to hyomandibular arch.
Ganglion.
*(VIII) Acoustic. Arises with (posterior portion) facial. Sup-
plies saccule and utricle of ear. Ganglion.
3. Just posterior to ear. Closely associated with each other.
(IX) Glossopharyngeal. Arises with vagus but sends branches
to first bra~chial arch. Ganglion.
(X) Vagus. Arises with glossopharyngeal but sends branches
to second, third, and fourth arches to viscera and lateral line
organs.

*IV. SPINAL CORD AND NERVE. Best examined in region posterior to pronephros.
a. Central canal. Expanded dorsal and ventral to a constricted
central portion.
I
b. Ependymal layer. Layer of cells lining central canal.
c. Mantle layer. Thicker layer of cell bodies expanding on either
side. Gray matter.
I
d. Marginal layer. Outermost layer of axons. White matter.
e. Dorsal root. Thin bundle of afferent fibers, running ventral to
join dorsal root ganglion.
f. Dorsal root ganglion. Elongate thick bundle of neurones on
either side of ventral half of spinal cord.
g. Ventral root. Thin bundle of efferent fibers, leaving ventro-
lateral angle of spinal cord to join nerve trunk.
h. Nerve trunk. Composed of efferent fibers from ventral root and
afferent fibers from ganglion.
i. Dorsal ramus. Branch from trunk to dorsal muscles.
j. Ventral ramus. Branch from trunk to ventral muscles.
k. Communicating ramus. Passes around notochord to join sym-
pathetic ganglion.
1. Sympathetic ganglion. A small mass of ganglion cells on either
side of dorsal aorta. Not a part of the spinal nerve but asso- II
ciated with it. -
 11-MM. EMBRYO 23
V. SENSE ORGANS OF THE HEAD
a. Ear. Now partially divided into two regions.
1. Utricle. Mesia! and dorsal portion.
Semicircular canals. Two: anterior dorsoventral canal, and
outer horizontal canal. (The third, posterior dorsoven-
tral has not developed.)
2. Saccule
* Endolymphatic duct. Posterior to level at which anterior
semicircular canal joins utricle. Between utricle and
brain. Joins saccule.
Cochlea. Heavily pigmented and ciliated area on inner
ventral angle of saccule.
b. Eye
* 1. Sensory layer of retina. Rods and cones, inner and outer gran-
ular layers and ganglion layer can be distinguished with high
magnification.
2. Pigmented layer. Still characterized by heavy pigmentation.
3. Lens. Solid, lying in mouth of optic cup.
4. Choroid. Thin vascular layer (formed from mesenchyme).
5. Sclera. Layer of mesenchyme investing choroid.
6. Corn(!a. Continuation of sclera between lens and skin.
7. Eyeball muscles. Small masses of muscle between eye and
brain.
c. Nose. The nasal pits have grown back to open into pharynx.
1. External nares. Open to exterior.
2. Internal nares. Open to pharynx.
3. Sensory epithelium. On side of tube nearest brain.

ENDODERMAL DERIVATIVES

The endodermal tube now opens to the exterior at both ends, by means of the ectodermal stomo-
deum and proctodeum respectively. The boundaries between ectoderm and endoderm at these points
cannot be traced. Commence at the mouth and trace the digestive tube and its derivatives back to the
cloaca! opening.
a. Mouth. Extends from the oral aperture to the point where the
internal nares mark the entrance of the nasal tubes.
b. Pharynx. The tube now flattens dorsoventrally to form the
pharynx.
c. Visceral (branchial) clefts. The pharynx opens to the exterior
at four points on each side where the visceral pouches (II, III,
IV, and V) have opened into the corresponding branchial grooves.
Pouch VI is vestigial and does not make contact with the ex-
terior. The operculum covers the external gills. The clefts do
not open directly to the exterior but ventrally into the opercular
cavity. The external gills are now degenerated and will be
replaced by internal gills. The opercular chamber approaches
the ear very closely.
* d. Thyroid gland. Beneath the floor of the pharynx. Completely
separated from it. The thyroid gland is Jlow dividing at the
posterior end. Pigmented and located just anterior to the heart.
24 ANATOMY OF FROG EMBRYOS

e. Thymus glands. Those from the first visceral pouch have dis-
appeared, those from the second visceral pouch have separated
from the dorsal end of the pouch. They are not easily to be
identified and may be omitted.
f. Parathyroid glands ( epithelioid bodies). Small masses of tissue
formed on the ventral ends of the third and fourth visceral
pouches. Their identification may be omitted.
g. Ultimobranchial bodies. Small pigmented bodies on the ventral
side of the sixth visceral pouch, at the level of the trachea. Iden-
tification may be omitted.

-------Oral cavity
(Posterior
nares) ---------

Oesophagus --

Ductus
- ,- choledochus
____ Pancreas

Lung· 'Stomach
'-Duodenum
,,,,. ...
Intestine ...

________ (Pronephric
Cloaca----------- ducts)

FIG. 12. Endodermal derivatives of 11-mm. Rana pipiens embryo. From a wax plate
reconstruction representing a cast of the digestive tract. Dorsal view.

* h. Trachea and lungs. A slitlike dorsoventral evagination from the

ventral side of the pharynx in the region of the visceral clefts.
This is the Laryngotracheal groove. After passing a few sections
~this groove separates from the oesophagus above, becomes a tube,
and sends out a branch, at sharp right angles to either side.
These are the bronchi. As each bronchus proceeds backward its
walls become highly vascular until at the end they condense
again at the growing point.
i. Oesophagus. From the region of the laryngotracheal groove the
oesophagus narrows rapidly and bends to the right at a level
behind the liver to enter the stomach.
j. Stomach. Easily recognized by its wrinkled lining. The stomach
may be traced forward and dorsally until it curves backward to
join the duodenum.
k. Duodenum. From the pyloric end of the stomach, the narrow
duodenum crosses over the oesophagus, swings to the left under
the hind-gut to join the coiled intestine. -
 11-MM. EMBRYO 25
l Liver. This large gland is now on the right side of the coelom.
It has grown around the vitelline veins and is highly vascularized.
On the dorsal side is the original liver diverticulum, now the
gall bladder. From the posterior end of the gall bladder the
common bile duct ( ductus choledochus) proceeds through the
mesentery connecting liver and stomach until it enters the duo-
denum on its anterior surface.
* m. Pancreas. This gland is found alongside the inner curvature
of the stomach. The pancreatic duct joins the common bile duct
shortly before the latter enters the duodenum.
n. Intestine. Following the duodenum, the intestine increases
greatly in diameter, and forms two complete spiral coils ventrally.
It then ascends as a much smaller tube coiling within the larger
spirals just described. On approaching the oesophagus it bends
sharply backward, rapidly decreases in diameter, and slowly
bends downward until it joins the proctodeum. There is no cloa-
cal swelling. Shortly after crossing the duodenum the intestine
receives the fused ends of the pronephric ducts.

MESODERMAL bERIVATIVES

1. MESENCHYME. In the head region condensations of mesenchyme have laid down the membranous
cranium iR which cartilage-forming (chondrogenous) centers have appeared. These give rise to the
neurocranium. (See page 29.) Connective tissue of various kinds and the blood and lymph vessels of
the head are also formed from the head mesenchyme. In the tail fin, mesenchyme is also present in
great amount. Elsewhere in the body mesenchyme fills in the spaces between organs not occupied
by body cavities.
11. NOTOCHORD. Still large and conspicuous. When examined with high magnification a thin layer of
mesenchyme cells (from sclerotome) may be found around it, and continuing up around the spinal
cord. This is the beginning of the axial skeleton, which lags behind the cranial skeleton.
111. SOMITES. The most anterior somites are disappearing. Specimens may have eleven or twelve in
the trunk, and a much larger number (thirty to thirty-five) in the tail.
a. Dermatome. This region of the somite is now broken up into
mesenchymatous cells which have made their way to the ecto-
derm of the dorsal half of the embryo where they will form the
dermis (cutis) of the skin. This layer apparently arises from
, somatic mesoderm in the ventral portion of the body.
b. Sclerotome. Now forms the mesenchymatous zone, from which
the axial skeleton will develop.
c. M yotome. The remainder of the somite is now converted into
muscle bundles lying close to the notochord. In the tail region
each is divided into a dorsal and a ventral bundle.
IV. INTERMEDIATE MESODERM
a. Pronephros. Greatly enlarged owing to the coiling of its
tubules. The nephrostomes are ciliated.
* b. Pronephric ducts. May be traced back on either side of the
dorsal aorta and dorsal to the posterior cardinals until they
unite. The fused portion bends ventrally and anteriorly to
join the narrow cloaca.
c. Mesonephros. Not yet commenced its'development.
26 ANATOMY OF FROG EMBRYOS

d. Gonad. Not yet developed. At the level of the pancreas it is

possible to find with high magnification primordial germ cells.
They are located in the roof of the coelom ventral to the pos-
terior cardinal veins and in the dorsal mesentery at its junction
with the coelomic roof.
Nephrostome I
I
I
I
,I

---- __ Pronephric
duet

----- __ Opening
./ into cloaca

FIG. 13. Excretory system of 11-mm. Rana pipiens embryo. From a wax plate recon-
struction. Ventral view.

v. LATERAL MESODERM. Somatic and splanchnic layers are closely opposed to the ectoderm of the ven-
tral body wall and the endoderm of the viscera respectively.
a. Coelom. Three regions may be distinguished.
1. Pericardial cavity. Still connects with the abdominal cavity
posterior to it.
* 2. Pronephric chamber. In the process of being separated from
the abdominal cavity beneath it by the growth of the lungs.
3. Abdominal cavity. Does not extend behind cloaca.
b. M esenteries. The dorsal mesentery is very thin but its two
layers are apparent at high magnification. The ventral mesen-
tery is represented only by the thin sheet between stomach and
liver (gastrohepatic omentum).
VI. crncuLATORY SYSTEM. Consists of heart, arteries, veins, and lymphatics. Red blood corpuscles now
easily distinguished.
a. Heart. Beneath pharynx. Commence at anterior end.
1. Bulbus arteriosus. First section of heart encountered.
* 2. Truncus arteriosus. Opens dorsally from bulbus. A short
vertical vessel. ·
3. Ventricle. Thick walled and muscular, the most ventral of
the two cavities seen in the same section. Communicates
with bulbus arteriosus. -
 11-MM. EMBRYO 27

4. Atria. Lie dorsal to ventricle. Partially separated into right

and left atria by interatrial septum. Both communicate with
ventricle through common *atrioventricular foramen. Left
atrium receives the pulmonary vein (see below) on dorsal
posterior surface.
5. Sinus venosus. Most posterior region of heart. Communi-
cates with right atrium. Receives common cardinal veins
(see below) at extreme right and left angles. Receives post-
caval vein from liver (see below) ventral to right common
cardinal vein.
ARTERIES
VEINS
Commissural- - - - - - ---·,
Anterior____ -------- \
cerebral \ Anterior
External --- --- \ - - - cardinal
carotid ',
Basilar --- ~l"::::.::::::-r

____ Inferior
.;'
,..'
jugular
Efferent,,,._ - _..!
branchials ..... ,
',-----
--- ~~.;:i, ___ Common
cardinal
Cutaneous ---~:;;:;;:;:'7"'2~
Pulmonary - -- -K+-0-~rl'?<i'---- Pulmonary
Glomus ----u-~"1,,..:.,-~..,~
--- Pronephric
sinus

Dorsal aorta------- ------Posterior cardinal

Fm. 14. Plan of circulation in 11-mm. Rana pipiens embryo. From a graphic recon-
struction. Dorsal view.

b. Arteries. Only the main arteries are to be identified.

1. Afferent branchial arteries. Commence at level of truncus ar-
teriosus. This vessel divides into left and right branches.
Each branch divides into two portions. The anterior portion
swings forward and laterally and then divides to form the
first and second afferent branchial arteries. The second branch
swings laterally and divides to form the third and fourth
branchial arteries. From the first branchial artery, the exter-
nal carotid (lingual) artery passes forward to the lower jaw.
2. Efferent branchial arteries. Locate the dorsal aorta at a level
just posterior to the liver. Running forward it divides into
the two *radices aortae (aortic roots), which run forward at
the lateral margins of the pharynx. Where the pharynx
begins to widen each root is joined by the fourth efferent
branchial artery. From the fourth efferent branchial artery
on each side ~ pulmonary artery grows out under the pharynx
to supply its lung, and a cutaneous axtery makes its way to
the skin.
28 ANATOMY OF FROG EMBRYOS

Just anterior to the point where the fourth efferent

branchial joins the aortic root, the third efferent branchial
can be found. Anterior to the point where the third efferent
branchial artery joins it, each root moves towards the mid-
line, dorsal to the pharynx. They are then joined by large
vessels, the second and first efferent branchial arteries, re-
spectively. Anterior to the point where the first efferent
branchial artery joins it, each root continues anteriorly and
dorsally on either side of the base of the brain, to become an
internal carotid artery. These arteries connect by a commis-
sure beneath the infundibulum of the brain and divide into
several smaller vessels of the head.
3. Dorsal aorta. From junction of aortic roots backward until
it enters the tail as the caudal artery. Gives off numerous
intersegmental arteries in its course.
* 4. Glomus. A vascular mass of tissue, supplied by branches
from aortic roots anterior to their point of junction. Sus-
pended in pronephric chamber median to pronephros on
each side.
5. M esenteric ar:tery. Large vessel leaving dorsal aorta on ven-
tral side and proceeding down dorsal mesentery to supply
intestine.
c. Veins. Usually have thinner walls than those of arteries.
1. Hepatic-portal system. Formed from vitelline veins after the
liver has grown around them.
Hepatic veins. Originally from liver to sinus venosus.
They are now connected with the postcaval vein, a very
large vessel (see below). From this stage on, the name
hepatic veins is applied to the vessels inside the liver,
whereas the portion from liver to sinus venosus is called
the postcaval vein.
Portal vein. Branches from pancreas, stomach, and intes-
tine unite to enter liver at posterior surface.
II

•
2. Cardinal system. The posterior cardinals now in process of
transformation to postcaval.
Posterior cardinal veins. Best studied by commencing with
caudal artery in tail. This divides into two vessels which

•
join posterior cardinals at level where pronephric duct
enters cloaca. They run forward ventral to dorsal aorta
and mesial to pronephric ducts. In some specimens, a
little further developed, these veins fuse.
* Postcaval vein (posterior vena cava). At the level where
the mesenteric artery leaves the dorsal aorta, this large
vein enters the dorsal surface of the liver. It can be
traced back to right posterior caval vein which it joins.
It runs forward through the liver to enter sinus venosus
ventral to right common cardinal vein.
NOTE: Anterior to point where postcaval vein unites with right common cardinal vein, the cardinal veins become much
reduced and can be seen only with high magnification. They follow the pronephrie duct of each side t.e enter the pronephric
sinus, a saclike structure partially enveloping each pronephros.
 11-MM. EMBRYO 29
~
(Jommon cardinal veins (ducts of Cuvier). Drain pro-
nephric sinus. Descend vertically across front surface
of pronephroi to enter sinus venosus at posterior lateral
angles.
Anterior cardinal veins (superior jugular). Join common
cardinals at their most dorsal level. Accompany aortic
roots into head. Finally take position dorsal and lateral
to internal carotid artery.
Inferior jugular veins. Join common cardinals at their most
ventral levels. Run forward on each side of heart. Ac-
company external carotid arteries to lower jaw.
d. Lymphatic system. Difficult to trace without assistance of in-
jected specimens.
Spleen. Thickening in wall of dorsal mesentery alongside mesen-
teric artery. Represents lodgment of lymph cells which will
later develop into spleen.
VII. CHONDROCRANIUM. The plan of the embryonic skull is laid down in connective tissue under the
brain, around the sense organs, and in the visceral arches. In this embryonic stage many cartilage
centers, easily recognizable by the large cells with tpeir relatively clear cytoplasm, may be identified.
The point of departure is the notochord, at the leyel of the anterior margin of the otic capsule, which
is still in the precartilage stage, but will later form a cartilaginous capsule around the inner ear.
"'LOWER JAW, HYOID. SKULL AND UPPER JAW
AND BRANCHIALS
--------Labial
-'t--11---- - - ---Cornu trabeculae
Mental---- - - - - - -
Meckelian------ ............. .

;,(.···
Ceratobranchial I_,
.· .
j.'.!I;,·:;.··--Iii~-,
.·"
Ceratobranehial II ___ _,/ /<' ..
/
Ceratobranchial m - - - - _,. - - - - - - - - - Para.chordal

FlG. 15. Chondrocranium of 11-mm. Rana pi-pi.ens embryo. From a graphic reconstruction. Dorsal view. Neurocranium outlined in
unbroken lines; splanchnocranium in dotted lines. (Non:: Pterygoquadrate, although formed from the first visceral arch, is outlined
in unbroken lines because it ia now fused with the neurocranium.)

a. N eurocranium. Around brain and sense organs.

1. Parachordals. On either side of the notochord at its anterior
end.
2. Basilar plate. Immediately in front of the notochord, united
with the parachordal on each side.
3. Trabeculae. These bars of cartilage swing wide to right or
left and then extend anteriorly on eitller side of basicranial
fontanelle.
30 ANATOMY OF FROG EMBRYOS

* 4. The basicranial fontanelle. This cavity is the seat of the

pituitary gland.
5. Pterygoquadrate (palatoquadrate). Joins posterior end of
the trabecula by its posterior ascending process, swings ven-
trally and to the side, and continues forward. This cartilage
is formed from the maxillary portion of the first visceral arch
but is mentioned here because of its union with the trabecula.
At a level in front of the eye is the vertical muscular process
for the attachment of jaw muscles. At the anterior end it
swings toward the median line and rejoins the trabecula by
its anterior ascending process.
6. Ethmoid (internasal) plate. Formed by the union of the tra-
beculae in the median line. Just anterior to the connection
between trabecula and anterior ascending process of the pter-
ygoquadrate.
7. Cornua tracecularum (trabecular horns). Two bars which
diverge at the anterior end of the ethmoid plate and continue
towards the anterior end of the head, accompanying the nasal
tubes.
8. Labial ( suprarostral) cartilages. Two short cartilages in the
upper lip, one on each side of the mouth. They meet the
cornua trabecularum.
b. Splanchnocranium. Formed in visceral arches. Follow from
front to rear.
1. Mental ( infrarostral) cartilages. In the lower lip on either
side of the mouth. Meet in the mid-line at their posterior
ends.
* 2. M eckelian cartilages. On the dorsal side of each mental car-
tilage and posterior to it. These rapidly extend to the side
where muscles can be seen connecting them with the muscular
process of the pterygoquadrate.
3. Ceratohyal. Several sections posterior to the last appearance
of the meckelian cartilage. This large cartilage is formed in
Ill
the hyomandibular arch.
4. Basibranchial (copula). An unpaired cartilage between the
two ceratohyals. The basihyal (anterior to the basibranchial)
has not yet developed cartilage. I ts position may be recog-
nized by the U-shaped thyroid gland which lies beneath it.
* 5.. Hypobranchials. Posterior to each basibranchial and cerato-
hyal and articulating with each. A platelike cartilage formed
in the third visceral arch.
6. Ceratobranchials. Extending back from the h}rpobranchials
as four slender processes. The most lateral of these cerato-
branchials, that is, the one in the third visceral arch, is the
first to be noticed as one goes through the sections from front
to rear. The second is in the fourth visceral, third in the fifth
visceral, and four th in the sixth visceral arch. The cerato-
branchials have not yet appeared in the dorsal portions of
the arches.
 PART II

ANATOMY OF CHICK EMBRYOS

THE GERM CELLS AND FERTILIZATION
The Ovum
The egg cell or ovum of the hen is the portion of the egg usually :referred to as the yolk. It con-
sists of a minute island of cytoplasm, the germ spot or germinal disc, floating on the yolk, which is a
large ball of stored-up ·food material. The whole egg cell is enclosed in a very thin, delicate vitelline
membrane. In addition to the egg cell proper, the egg, as we ordinarily know it, is made up of several
layers of albumen surrounding the yolk; and these in turn are enclosed in tough shell membranes lining
the hard calcareous shell (Fig. 16).

- - - Vitelline membrane
,_.._ _ Chalaziferous
layer of white
Shell
... Outer shell membrane
Inner shell membrane
Air cell

Chalaza

Dense white
_,____ Inner thin white
2 ' - - - Outer thin white

...____ _ _ _ White yolk

' - - - - - - - Concentriclayera
of yellow yolk
FIG. 16. Diagrammatic vertical section of a fresh hen's egg. From "PoultT11 Productinn," courte&fl L. E. Card, and the publi&hers,
Lea and Febiger.

The yolk is laid down in concentric alternating light and dark layers around a central mass of white
yolk called the latebra. This extends \ipward to the surface as a stalk of white yolk; where it spreads
out as the isthmus of Pander, beneath the germinal disc. The innermost layers of albumen become
twisted into coiled strands, the chalazae, which extend out from the yolk toward the ends of the egg.
The parchmentlike shell membranes are separated from one another at the blunt end of the egg, thus
forming the air chamber.
The albumen, shell membranes, and shell are membranes added to the egg during its passage down
the oviduct. The albumen is a source of protein food material for the growing embryo, and the shell
membranes and shell have a protective function.
It is interesting to note that a great deal of the stored-up yolk material is still present at the end of
the period of incubation, when it is resorbed; and hence it is available as nutriment for the newly
hatched chick.
Eggs vary greatly in size and in coloration, but the yolk is about 40 mm. in diameter and the germi-
nal disc about 4 mm. ·
ll

't
·' /
~
I
32 ANATOMY OF CHICK EMBRYOS

The jpermatozoon
The male germ cell or spermatozoon, in contrast to the female reproductive cell, is a very minute
body of microscopic size. It consists of an elongate head and a long, slender, filamentous tail which
merges almost imperceptibly with the head (Fig. 17). The head, which is made up largely of nuclear
materials, is surmounted by a slightly recurved acrosome.
Fertilization
The ovary of the laying hen contains numerous ova in various stages of development.
The rate of growth of the vast majority of them is very slow and they are very small in con-
sequence. In succession, certain ova, a few at a time, begin to increase rapidly in size) so
that in the last 5 to 8 days of development fully 99 % of the yolk material is added to each
egg. Then, when the maturing egg has completed its growth, it is set free from the follicle
of the ovary in which it developed and is picked up by the funnel of the oviduct. Imme-
diately after it is liberated into the oviduct it is surrounded by fluids which contain myriads
of spermatozoa received from a male in a prior copulation. Some of these enter the egg cell,
and one fertilizes it. The remainder migrate to the periphery of the germ spot where they
ultimately disintegrate.

CLEAVAGE AND GERM LAYER FORMATION

FIG. 17. Sper- The processes of cleavage and gastrplation in the hen's egg take place during the pass-
matozoon of
the fowl. age of the ovum down the oviduct. Cleavage, from its earliest stages, is quite irregular and,
si,pce it involves only the tiny island of cytoplasm lying on the yolk, a blastula is soon formed
consisting of a thin, discoidal sheet of cells separated from the yolk by a fluid-filled cavity, the blastocoel.
The process of gastrulation follows immediately after cleavage and results in the formation of a two-
layered embryo. The exact manner in which this is brought about is very difficult to demonstrate.
Until recently it was considered to be a process of invoJution whereby the cells along a particular portion
of the margin of the blastodisc (the dorsal lip of the :Blastopore) rolled under the original layer. Lately
it has been shown that the process involves polyinvagination, that is, numerous small grooves appear on
the surface of the blastodisc and at these points cells insinuate themselves beneath the surface layer. 1
·From these first migrants, strands of cells are produced which spread out and coalesce to form a con-
tinuous sheet of endoderm beneath the original surface layer which is now ectoderm (Fig. 18).

a b c d
FIG. 18. Cleavage and endoderm formation in the chick. a, b, early cleavage ; c, endoderm form ation by polyinvagination: small
solid lines represent furrows along which future endoderm (stippled) invagimttes; d, the completed sheet of underlying endoderm
(stippled) .
Mesoderm Formation
When development is resumed during incubation, the blastoderm begins to grow out rapidly over
the yolk and formation of mesoderm from the two-layered gastrula begins. This involves an interme-
diate step, namely, the formation of the primitive streak. This structure has the appearance of a thick-
ened line on the blastoderm. Until recently it was considered to be due to concrescence, a process
involving the forcing together of the two halves of the dorsal lip of the blastopore. Actually it appears
to be formed by c ence of surface material toward the future primitive streak accompanied by com-
plicated streaming movements of cells in certain surface areas. 1 The whole effect, therefore, is to force
1 A. M. Dalcq, "Form and Causality in Early Development," Cambridge University Press, 1938.
, -tr 1· • I .r
2• ~ v a;t, }- ~ 'L AQ ~ M- ' ~r\..t. p ~~ ., _, o-
A.-~ ~ c.f.,__~ ~ tt- J)o ? 1 ~ ) , - -r ~ owe r
I
 24-HOUR EMBRYO

together cells from the right and left sides of the blastodisc as a linear thickening and to allocate certain
cell groups to special regions of the blastoderm where they are destined to give rise to particular parts
of the embryo. Some of the cells which move to the primitive streak invaginate and spread out between
the two layers thus forming the mesoderm (Fig. 19). It grows out on each side from the tissue of the
primitive streak as a broad winglike sheet extending forward and laterally. A rod of cells which consti-
tutes the notochord also grows forward from the anterior end of the primitive streak, and is, therefore,
considered to be of mesodermal origin.

a b c d
Fm. 19. Formation of the primitive streak and mesoderm in the chick. a, the gastrula ; b, convergence of surface cells toward future
primitive streak; c, d, lateral mesoderm and notochord extending outward from the primitive streak. Mesoderm heavily stippled,
primitive streak solid black. ,.-- 1 1
~'i r ~

CLd ctJ
18-HOUR EMBRYO

After 18 hours of incubation the chick embryo·begins to take definite form. The early part of this
-
period is occupied by the process of gastrulation and the formation of the third germ layer, or mesoderm.
Consequently, at this stage the blastoderm has the form of a small round plaque of tissue about 1 em.
in diameter. In this blastoderm there can be seen a general division into a small, clear, inner area, the
area pellucida, and a broad, outer, heavily stained margin, the area opaca.
The following parts of the embryo can be distinguished:
Head fold. A heavily staining crescentic fold of blastoderm near one end of the area pellucida.
Primitive streak. A heavily staining thickened streak tailing off away from the head fold toward the
opposite side of the area pellucida. The primitive ridges constitute the lateral margins of the streak,
and between these is seen the sunken primitive groove. The anterior end of the streak is marked by
the nodelike primitive knot.
Notochord. A rodlike structure composed of mesoderm extending forward from the primitive knot
toward the head fold can be seen in favorable specimens.

Laboratory Directions
Study carefully a stained total mount of an 18-hour embryo and identify the structures described
above. Make a drawing of this embryo 2 inches in diameter and label all parts (page 34).

24-HOUR EMBRYO

BoDY FoRM

In the 6-hour interval of incubation which has elapsed since the 18-hour stage, considerable change
has occurred in the embryo: .
The head fold has become much more prominent and appears as a flattened finger-shaped struc-
ture protruding forward from the blastoderm, beginning at the level of the anterior intestinal portal
(Fig. 20). The body is well marked out, and a number of paired blocklike somites have been formed on
each side of the mid-line in the space between the head fold and the primitive knot.
34 ANATOMY OF CHICK EMBRYOS

Total Mount of an 18-Hour Chick Embryo.

The blastoderm surrounding the chick shows several characteristic developments:

The area immediately surrounding the embryo, which is somewhat slipper shaped in general out-
line, is lighter and clearer in appearance and forms the area pellucida. Beyond the area pellucida the
blastoderm is more opaque and hence is called the area opaca. In this region, particularly where it

Notochord-

Fro. 20. Diagrammatic lateral view of the head fold of a 24-hour chick. Arrow indicates anterior intestinal portal.

adjoins the area pellucida, the blastoderm is splotchy in appearance owing to the development of heavily
staining scattered aggregates of cells. These are blood islands, and the whole region is known as the area
vasculosa. It is terminated laterally by a definite margin which later forms a blood vessel, the sinus ter-
minalis. Beyond the sinus terminalis the bla~toderm continues as the area opaca· externa.
c J, -
;( ; r

C@i

... --...
]11.:s_I >-• ..-
:.....,.....
" .. .:... .. :
1 ..· •

/ :.-:-: ••;'-.;.,.;.t,;..
,_ :,>.......... :~~-: ' rp.,.,·...,;f ,' ... e.
1~;,. e ~t
,...
I ·,
.
.... ~,,.. ..
.
.•.
• ... ' ,
- "Ri~t e.

,. ' ..··~
__"?.. ;'""it••t. 8r0111t-
- X. - i ... •,
~··
·-:;-:~·.­
·:-..,..._:
_,.. .-. ...
:l' . '
' :: .- .. .~, !>- - -
Jt'ofc Chl>-d •
.. ~

, -. , ...... .,.. /'/e~J f.IJ

\ .... ~
...
\ "l
\ -.... ,·.
.•.
-: . . ,I
\~

''
....
. ·,,. ,

F,·g I. 1:.lal 1'1ttu I "' If.It,. CJ,:~k

!m b..-., " :n.s (I. )

.-.,.:.
• r

-- '""·- -.... - -An. fr uo

r: I/ 1

...
..
-- ... '.;)'.
" . \.•
.·.
--.-~--- t>f/,'c ~"'.S•
.... :~·
c,

le,
- - - !lei1 J?, J
'· ...
l.

~ --
...· .... _ -·t-.._ _. Ho J
- f. !••

.;.- -
'-•

11 ) t•V •
Te.-........ 1·.s.
..
fy,'l'I'
--'~--
·~:-
,_:

...
"t. '
;;_

..
.,. .· .. ?

ti
) '
 36 ANATOMY OF CHICK EMBRYOS

ECTODERMAL DERIVATIVES

The ectoderm which forms the outer covering of the dorsal side of the head fold has already begun
to differentiate into the beginnnings of the neural tube. This is brought about by the development of a
furrow extending longitudinally down the dorsal side of the body. This furrow is bounded laterally by
two ridges, the neural ridges, and the depression between is the neural groove. As the depression deepens
the neural ridges ultimateiy meet in the mid-dorsal line, thus forming'a closed tube, the neural tube.
This process beg~ns l~ the anterior end of the head fold an~ gradually works backward, s? that _at ~hours
the neural tube 1s· ~ closed as far back as the postenor end of the head fold. Behmd th1s pomt the
neural ridges gradually open out so as to form the boundaries of the original neural groove. This process,
which involves the sinking-in and closing-off of the tissues in a certain area, such as the surface ectoderm
in this particular case, is known as invagination.
It is to be noted that at this stage the anterior end of the neural tube still retains a small opening to
the outside which is known as the anterior neuropore. Some differentiation may already be apparent in
that portion of the neural tube within the head fold to the extent that the anterior end of the tube, the
forebrain, has prominent lateral bulges indicating formation of the optic vesicles.

ENDODERMAL DERIVATIVES

At this st"age the endoderm, which lies next to the yolk, protrudes up into the head fold in the form
r of a flattened fingerlike pouch constituting the fore-gup. This lies beneath the brain, which is developing
as described above. The fore-gut is a tubular structure communicating with the mid-gut by way of the
anterior intesti.pal portal (Fig. 20). The mid-gut is open ventrally over the yolk, and lacks a floor.

MESODERMAL DERIVATIVES

At this stage the notochord extends forward from the anterior end of the primitive streak, beneath
the neural tube, and up into the head fold in the form of a solid, cylindrical rod. Its anterior end fre-
quently appears to merge with the neural tube. The lateral sheets of mesoderm which extend alongside
the notochord do not extend into the head proper but instead proliferate a delicate netlike tissue, the
mesenchyme, which grows up into the head and occupies the space between the developing organs already
present. The lateral sheets do not meet beneath the head fold, and consequently there is no mesoderm in
this region, which is known as the proamnion. In the region of the body proper, the portions of the
lateral sheets adjacent to the notochord become divided off transversely into paired blocks known as
somites. Little differentiation in the individual somite _i~ evident at this stage beyond the fact that a
cav· , the ~el, is present in the center of each. Six pmrs of somites are usually present at this stage.
Lateral to the somites the mesoderm is separated tangentially into two sheets of cells; the uppe · con-
tact with ectoderm is the somatic mesoderm, and the lower in contact with endoderm is the splanchnic
mesoderm. In the splanchnic mesoderm, cell aggregates forming the blood islands are to be found.

Laboratory Directions
A. Study carefully a stained total mount of the 24-hour chick and make a drawing of the embryo
showing detailed structure in the body proper and in a narrow strip of the blastoderm. The body should
be drawn approximately 4 inches long. Label all parts.
B. Study through the transverse serial sections supplied and make a set of drawings of sections cor-
responding to the following levels: (1) forebrain, (2) anterior intestinal portal, (3) a somite, (4) primi-
tive streak. Label all parts.

--
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I
 24-HOUR EMBRYO 37

(1) Section through the forebrain.

(2) Section through the anterior intestinal portal.

(3) Section through a somite.

( 4) Section through the primitive streak.

Transverse Sections of a 24-Hour Chick.

38 ANATOMY OF CHICK EMBRYOS

33-HOUR EMBRYO I
BODY FORM

After 33 hours of incubation the embryo has increased in size to a little over 4 mm. The head fold
is considerably longer and bends downward slightly over the anterior end of the notochord. The body
proper has also increased in length, and at this stage about 12 pairs of somites are present. Thickened
ridges, extending out laterally just in front of the anterior intestinal portal, mark the beginnings of the
vitelline veins. In the area vasculosa the blood islands are very prominent, and there is some indication
that they are beginning to coalesce to form the blood vessels of the yolk sac.

EcToDERMAL DERIVATIVES

The developing neural tube has closed over throughout almost its entire length. The anterior neuro- I
pore has usually been obliterated, but at the extreme posterior end the neural folds still remain broadly .
open, producing the large rhomboidal sinus. The brain shows three prominent vesicles at the anterior I
end, which named in order posteriorly are as follows: the prosencephalon, mesencephalon, and rhomben-
cephalon. Behind the rhombencephalon a varied number of small constrictions are apparent. The sides
of the forebrain or prosencephalon show prominent lateral bulges, which are the optic vesicles. On the
sides of the head fold alongside the rhombencephalon the auditory placodes.form thickened ectodermal
areas which later invaginate to become the otic (auditory) vesicles.
II
ENDODERMAL •DERIVATIVES I
Little change has taken place in the endodermal derivatives except that the fore-gut, extending up
into the h~aafold, has grown in length along with the head fold .
... ~ ;,t._ "'t MESODERMAL DERIVATIVES I
The notochord is still present as a median rodlike structure beneath the neural tube. The lateral
sheets of mesoderm ,are blocked off into twelve pairs of somites alongside the notochord. Mesenchyme
is present in the head as at 24 hours. -
In the region of the anterior intestinal portal the splanchnic mesoderm lying alongside the lateral
walls of the intestinal portal bulges out laterally into the body cavity so as to leave a narrow space be-
tween it and the endoderm. This is occupied by a delicate tube formed from cells proliferated off the I
adjacent splanchnic mesoderm. In front of the anterior intestinal portal these two tubes fuse, and ulti-
mately the dividing wall between them breaks down. In this manner the inner lining or endocardium
of a single-chambered, tubular heart is formed. In a similar manner the thick, overlying bulges of
splanchnic mesoderm also meet and fuse above and below this tubular structure so as to form the thicker I
epimyocardium of the heart and the supporting dorsal and ventral mesocardium. It will be noted,
therefore, that the heart arises from two fused primordia, and consequently the vessels which grow for-
ward from the heart are also paired. · I
These tubes growing forward from the heart are the ventral aortae. They are located beneath the
fore-gut, and as they grow forward from the heart swing dorsally around the anterolateral ends of the
fore-gut as the first aortic arches. They then course backward, dorsal to the gut, as the paired dorsal I
aortae. The actual extent to which this development has progressed at 33 hours varies with individual
specimens, but at least the heart and vitelline veins are uniformly present.
Laboratory Directions I
A. Study carefully a stained total mount of the 33-hour embryo. Make a drawing stressing par-
ticularly the form of the brain and also what can be seen of the developing circulatory system.
B. Study through the series and make drawings as follows: (1) through the region just in front
of the anterior intestinal portal showing the heart; (2) through the region of the anterior intestinal por· I
tal to show the vitelline veins. -

'
c "] ' If

··· ..-
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- - -- "tt1 '1h~ I 1 E!.

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-
 33-HOUR EMBRYO 39

(1) Anterior end of 33-hour phick showing heart and forebrain.

(2) Section through the heart.

(3) Section through the anterior intestinal portal and vitelline veins.

33-Hour Chick Embryo.

40 ANATOMY OF CHICK EMBRYOS

48-HOUR EMBRYO

EXTERNAL FORM
(,) ,.,
From the 33- to the 48-hour stage the body of the chick has increased greatly in size, and its shape
has been altered profoundly. The increase in size is due largely to the growth of the head and the addi-
tion of more somites to the body, so that its length is now about 7 mm. and the number of somites ap-
proximately 26. The change in form is brought about by the development of several flexures or bends
in the head fold and by twisting or torsion in the body proper so that the anterior portion lies mostly on
its left side. As a result of this change in form, the head is more readily accommodated in the small
space available beneath the shell since it lies flat on the yolk. At this stage the cranial flexure, which
usually begins to appear in the region of the midbrain at 33 hours, has become so pronounced that, taken
inconjunction with the cervical flexure in the region of the neck, the anterior end of the head is directed
backward. Twisting or torsion occurs at the level where the head fold rises from the blastoderm.
Hence the posterior half of the body lies in its original position with the ventral side down. At he ex-
treme caudal end, the tail fold is being marked out very much in the same manner as that by which the
head fold originated.
The head at this stage exhibits a number of prominent bulges corresponding to the divisions of the
brain. There are also present the beginnings of the sense organs-nose, eye, and ear. Just posterior to
the head on the lateral walls of the head fold three transverse slitlike openings can be seen. These mark
the pharyngeal region. Protruding out ventrally from the >
body
.
at about this level is the prominent heart.

Amnion and Chorion (Fig. 21)

The anter~t._part of the body of the chick is covered by a fold of somatopleure (ectoderm somatic +
mesoderm), which is thrown up over the body, beginning first in front of the head fold in the region
recognized earlier as the proamnion. Mesoderm, however, has grown into this region in the meantime.

Fm. 21. Development of the amnion and chorion in the 48-hour chick.

Since the fold is reflected back out over the yolk sac again, two independent coverings are formed,
namely: the amnion, which lies next to the embryo, and the chorion outside the amnion. The amnion
contains a cavity, the amniotic cavity, which becomes filled with amniotic fluid. The space bet-ween the
amnion and chorion represents an outward extension of the coelom called the exocoel.

Laboratory Directions
A. Study carefully a stained total mount of the 48-hour chick, and make a drawing of the embryo
showing detailed structures. The body of the embryo should be drawn approximately 6 inches long.
Label all parts.
CIRCULATORY SYSTEM

The circulatory system of the 48-hour chick represents a feature of fundamental importance. It
should, therefore, be studied with the greatest care in both injected total mounts and in serial sections.
The system is of mesodermal origin. Most of the details described below can be readily identified in in-
jected specimens, and after they have been worked out should be recorded by a drawing.

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 --~·

42 ANATOMY OF CHICK EMBRYOS

HEART

As pointed out in the study of the 33-hour chick, the heart develops from paired primordia, the
posterior ends of which represent the stumps to which the vitelline veins from the yolk sac will ulti-
mately attach themselves. The tubular structure lying in front of the anterior intestinal portal grows
much mot~ rapidly than the space in which it is accommodated. It adjusts itself to this situation by:
(1) Bending outwiJ,rd tu the right (as at 33 hours) (Fig. 22).
(2) By looping so that the posterior end is pushed up under the anterior end (as at 48 hours)
(Fig. 22).

-<;.e
.,..,1\'lt"
33 Hour 48 Hour 72 Hour
Frn. 22. Stages in the developm~t of the heart of the chick.

Furtherm~re, a certain amount of differentiation in the character of the walls of the chambers of the
heart has occurred at this stage so that we may recognize:
Sinu_s __ Ve_ri_o_sus. The thin-walled posterior end sitmi.ted,_ in the mid-line which serves as a general
receiving chamber. This region receives the fused vitelline veins, and hence "can be readily recognized by
locating them alongside the anterior intestinal portal and tracing them, after they have fused to become
the meatus venosus, forward into the heart.
A,tdum. Thin-walled and receiving blood from the sinus venosus. This chamber lies anterior to the
sinus~~s but ~;i~gs posteriorly to join the ventricle.
Ventricle. Thicker-walled with a developing reticulum of muscle bands, lined by a delicate endo-
thelium and receiving blood from the atrium. The ventricle constitutes the posterior portion of the loop
formed by the heart.
BulQ_us. Arteriosus·. Thi~ed, roun,d or oval in cross-section and possessing a delicate endo-
thelium. This chamber runs forward from the ventricle and swings toward the mid-ventral line just
below the visceral arches and clefts of the pharyngeal region.
Ve.ntril _Aorjae. The blood leaves ,the bulbus arteriosus by way of the ventral aortae, which at this
stage have almost ceased to exist, owing to their failure to keep pace with the growth of related vessels.

ARTERIAL SYSTEM

4J2rtfr:_{irc;hes. From the ventral aorta the blood was originally carried to the dorsal aorta by means
of paired vessels swinging around the front end of the fore-gut and constituting the first aortic arch.
These vessels still persist as the paired first aortic arch flowing thr9ugh the first visceral arch, which has
been established by the demarcation of the first visceral cleft. In addition, -paired aortic arches have de-
veloped in the second and third visceral arches. These are the second and third aortic arches, respec-
tively. They also course upward from the ventral aorta to join the paired dorsal aortae.
External Carotid. A minute vessel located at the point of origin of the first aortic arch. It can
sometimes be seen to extend into the base of the first visceral arch where it breaks up into a capillary
network. -
 48-HOUR EMBRYO 43

Internal Carotid Arteries. From the point where the first aortic arch meets the dorsal aorta, the
latter extendsforward into the head as the internal carotid artery to sup£!Y-~__capillary network to the
brain. -
.-- Dorsal Aortae. The dorsal aortae are still paired in the region of the pharynx, and each vessel is
often spoken of as a "radix" of the dorsal aorta. At about the level of the heart they fuse to form a
single dorsal aorta. This opens out again to the paired condition slightly anterior to the level at which
the vitelline arteries flow out to the yolk sac.
Vitelline Arteries. These course out laterally over the yolk sac about the level of the twenty-
second somite.
Caudal Arteries. The backward prolongations of the dorsal aortae beyond the vitelline arteries
form the caudal arteries.
lntersegmental Arteries. Since the body is growing rapidly by the lengthening of the head fold
and the addition of somites to the trunk, a series of intersegmental arteries develop to supply its nutri-
tional needs. These vessels are paired and course outward and dorsally between the somites arising from
the dorsal aorta (or aortae).

VENOUS SYSTEM

A. SPLANCHNIC VESSELS: VITELLINE VEINS

The blood distributed to the yolk sac by the vit~lline arteries is returned to the heart by the vitel-
line veins. The latter converge toward the region o~ the anterior intestinal portal, where the two main
trunks fuse to form the meatus venosus just before entering the sinus venosus. The vitelline arteries
and veins anastomose
.... freely over the yolk sac, but large trunks from both arteries and veins communicate
with the sinus terminalis encircling the blastoderm.
B. SOMATIC VESSELS: THE CARDIN AL SYSTEM
Since a great increase has occurred in the amount of blood supplied to the developing body, a new
set of veins, the cardinals, develop to drain blood back to the heart. These are laid down following the
general plan of the letter H-modified, of course, to the contours of the body.
Anterior Cardinal Veins. These vessels drain the head arising from the capillary plexus of the in-
ternal carotids over the brain. They course backward, following the ventrolateral angles of the neural
tube until they have just passed the level of the otic vesicles. At this point they begin to swing ven-
trally but pass lateral to the dorsal aorta to join the common cardinals.
Common Cardinal Veins. These vessels (also paired) receive blood from the anterior and posterior
cardinals and empty into the dorsolateral angles of the sinus venosus.
Posterior Cardinals. These vessels drain the somites of the posterior end of the body coursing lat-
eral to the dorsal aortae and in close association with the developing mesonephroi, or embryonic kidneys.
lntersegmental Veins. The blood distributed by the intersegmental arteries is returned to the car-
dinals by way of the intersegmental veins which lie between somites and anastomose with the interseg-
mental arterioles.
NOTE: One of the most outstanding features of the developing circulatory system is the fact that all primordia
of the original system are paired and that practically all new additions to the system are also paired. Future develop-
ment depends on elimination of certain parts of the system and general simplification.

Laboratory Directions
B. Using injected 48-hour total mounts, study the circulatory system carefully and make a drawing
showing the vessels described above. Label all parts.
 ANATOMY OF CHICK EMBRYOS

I
I
Circulation in 48-Hour Chick Embryo-Injected Specimen.
 48-HOUR EMBRYO 45

REPRESENTATIVE SECTIONS OF THE 48-HouR EMBRYO 1

A. Section through the upper part of head (Fig. 23)

This section was taken through the head at a level just below the floor of the brain in the region
of the cephalic flexure. For this reason it shows the mesencephalon (9) as a thick-walled rounded struc-
ture quite separate from the thin-roofed myelencephalon ( 1). In the mid-line between the two is a
small portion of the notochord ( 4). Alongside the left ventrolateral margin of the myelencephalon is a
heavy condensation in the mesenchyme (2). This is the semilunar ganglion of the trigeminal nerve
(V). The nerve proper (3) can be seen leaving the lower side of the ganglion. The other trigeminal

-16

I
3
Fxa. 23. Section through the head of a 48-hour chick at the level of the mesencephalon.

nerve (11) has been cut so that it appears as a small band of fibers in the mesenchyme on the opposite
side. The anterior cardinal veins (12) also lie along the ventrolateral margin of the myelencephalon,
and the one shown on the right side (10)" extends forward toward the mesencephalon. Small branches of
this vein (13) are located alongside the myelencephalon. Closely applied to the surface of the mesen-
cephalon are a number of small blood vessels (5, 6) which represent portions of the capillary plexus of
the internal carotid artery supplying the brain. Surrounding the head are: the amnion (15), which
forms a closed sac; the chorion (14), which lies outside the amnion; and the yolk sac (16), also covered
by the chorion and containing numerous blood vessels (17). ·The amniotic cavity (8) and the exocoel (7)
are also shown.
B. Section through the upper end o£ aortic arch I at the point of origin o£ internal carotid arteries
(Fig. 24)
In this section the head is cut twi((e, owing to the cranial flexure, and the posterior end is almost
completely separated from the anterior end. In the posterior end note the thin-roofed myelencephalon
(1), alongside which lie the otic vesicles (2, 20). Owing to the tilt in the plane of section, the left (2)
appears to be just invaginating but the right (20) appears as a closed vesicle. Associated with tne left
otic vesicle is the acoustic ganglion (3) of the auditory nerve VIII. The anterior cardinals (19) lie at
the ventrolateral angles of the myelencephalon; the notochord ( 4) is ventral to it. Immediately above
or dorsal to the gut are the paired dorsal aortae (18).
In the anterior portion of the head the brain is represented by the thick-walled, oval-shaped dien-
cephalon (12) which is constricted ventrally to form the infundibulum (13). Beneath the infundibulum
is Rathke's pocket (9), which represents an evagination of the stomodeum (8), and which, in associa-
tion with the infundibulum, forms the hypophysis or pituitary body. Extending alongside the stomo-
deum is a pair of elongate blood vessels. These are the internal carotids (16) and the first aortic arch
1 All sections are shown exactly as they appear in the microscope. Descriptions apply only to the sections illustrated in the

manual. The student will probably encounter some variations in the sections studied by him. -
 ANATOMY OF CHICK EMBRYOS
I
46

( 17) combined. Small branches of the carotids ( 11) lie close to the brain. Branches of the anterior
cardinals (10) are evident in the lateral areas. The right optic cup (14) is cut so that it shows as a
I
hollow structure containing the solid lens primordium ( 15). The pharynx shows two lateral outpocket-
15
l
I I
I
-22

I
I
I I
6 5
F10. 24. Section through the head of a 48-hour chick at the level of the eye and ear.
II
ings which form the first visceral pouches (6). Atlterior: to the first pouch on the right side, the head
shows a distinct bulge which represents the first visceral arch (7), in which the first aortic arch flows.
The first viseeral groove (5) is to be seen on the side of the head.
C. Section through the body proper and heart (Fig. 25)
At this level, and throughout the rest of the series, four typical structures are to be seen in the mid-
line arranged from dorsal to ventral, as follows: spinal cord (1), notochord (2), dorsal aorta (3), gut
(4). Lateral to the spinal cord are paired thickenings in the mesoderm constituting the dermatomes (14)
I
,...1s
/ I

I
I
l
6
Fm. 25. Section through the heart of a 48-hour chick. I
of a somite. Lateral to the gut are two cavities which represent forward extensions of the coelom (13).
The left, in this section, communicates directly with the exocoel. In the ventrolateral extensions of
the body wall are large paired blood vessels, the common cardinals ( 12), and of these the right com- I
municates with the sinus venosus of the heart. The parts of the heart are arranged in order forming
a loop and marked off by slight constrictions, as follows: sinus venosus (5), atrium (6), ventricle (7),
bulbus arteriosus (10). Note the thin endothelial.lining (9) of the bulbus and ventricle contrasting
with the thicker outer epimyocardium (8).
I

•
 48-HOUR EMBRYO 47

D. Section through the mid-body at the level of origin of vitelline arteries (Fig. 26)
· At this level the thick-walled neural tube (10) which lies below the surface ectoderm is somewhat
elliptical in outline. The notochord lies beneath the neural tube. The dorsal aorta (3), paired at this
level, lie beneath the notochord but slightly lateral in position. The vitelline arteries (1) course out-
ward laterally from them. The endoderm of the wide-open mid-gut (2) forms the basal layer. Lateral
to the neural tube are the paired somites. In these we may distinguish a 1:ieavy platelike dermatome

'
'l
FIG. 26. Section through the body of a 48-hour chick at the point of origin of the vitelline arteries.

(11) closely associated with the surface ectoderm, the poorly defined sclerotome (8) lying next the
spinal cord and notochord, and the myotome (9), or somite proper, forming an inner hook on the der-
matome. The nephrotome has already differentiated into the nephric duct (5) and tubules ( 4) ; the
duct being thick-walled and circular in outline while the tubule is ventromedial to it and irregular in
section. The lateral sheets of mesoderm are divided into the upper somatic layer, which is associated
with the ectoderm to form the somatopleure (7), and the lower splanchnic layer associated with endo-
derm to form the splanchnopleure (6). The exocoel (12) lies between these layers.

E. Section through base of tail fold (Fig. 27)

The tail fold has begun to protrude above the surface of the blastoderm in a manner similar to
that of the head fold. Into it. a blind extension of the endodermal tube, the hind-gut (3), will extend,

''
1
Fm. 27. Section through the tail fold of a 48-hour chick.

and at this level the posterior intestinitl portal, which represents the point of communication with the
mid-gut, is to be seen. The bulk of the tail fold is made up of a solid mass of mesodermal cells ( 4, 5).
Ectoderm (6), somatic mesoderm (7), splanchnic mesoderm (1), and endoderm (2) are clearly recog-
nizable.
Laboratory Directions
C. Study the serial sections provided and find, draw, and label sections illustrating the follow-
ing levels:
(1) Section through the head showing developing eye or ear (Fig. 24).
(2) Section through the heart (Fig. 25).
(3) Section through mid-body region showing the amniotic raphe (which is the level at which the
lateral folds of the amnion meet above the embryo).
(4) Section through the tail fold (Fig. 27).
48 ANATOMY OF CHICK EMBRYOS

':Dorsa./ Aorl• - , 1--ltrifeY'11r l'u~11111 \}ej"'

Sro • d '''"' ·,
/,st 1ttit r• ~J I :- tr'Je• •:i
J_

/1uc. c .... ri -'

::!) IC r
r,wr.11 •.,
°f'llc c .. r- _- /. 01tsti c
....
o.fA .. ,+, ...~
~
I
I ---- Kofo~li o rd
"'Ct\5 - - - - -
,~ ---- _ _ • _ Is~ Vo'rc.e r...
______ _ )sf Vise ~;a. I (}.,.,. >'C..
_ _ }s-t /)jtt er ii I

....
(1) Section through the head.
I
c ... • Q. ,..:r-
- 1
I

I
s;,, ... s Ve
I
e
'/

(2) Section through the heart.

48-Hour Chick Embryo.

 48-HOUR EMBRYO 49

e.

A""" n• •O\ f'

A ,., , , ol 1 c • "' )
_____ ;Yo-/ c u

Io (J- ,:ii />"" ,I..._ ,..-{_

(3) Section through mid-body region.

""'' .....
,- - --
' \
......... '

c
I o el"'rn o-1 s <!..
~ .. d-Gv..:~
t

( 4) Section through the tail fold.

48-Hour Chick Embryo.

 50 ANATOMY OF CHICK EMBRYOS

STUDY OF SERIAL SECTIONS

Methods. It should be quite obvious, from the work already done in drawing and identifying
structures in a few representative sections of an embryo, that a thorough knowledge of its anatomy and
the relations of various parts can never be obtained by such a procedure. A set of serial sections con-
sists of a series of slices taken in order through the embryo from the tip of the
- . . x head to the end of the tail. Consequently every structure in the embryo will be
represented in a certain group of sections in some particular part of the series.
c As a result, it is possible to build up a mental picture of any particular structure
, A and its relation to surrounding structures by tracing it, forward and backward
in the series, and studying it independently of, or in relation to, associated
structures.
It is highly desirable, therefore, that the student should now attempt to sub-
stitute this method of study for the time-honored process· of making drawings,
except, for example, in the case of the total mount. The drawings already made
have served a useful purpose in making the student familiar with the general
appearance and structural plan of such organs as the neural tube, gut, and blood
vessels.
. y'
Orientation of the Sections (Fig. ~8). Before beginning to study a series of
sections it is always advisable to try to get some conception of the plane of sec-
Fm. 28. Diagram illustrat- tion ABCD in relation to th~ iongitudinal axis of the body x-y. It should be
ing the relation of the realized that no two series are absolutely identical because the plane of section
plane of section ABCD to
the longitudinal llXie of the may be tilted in many ways from left to right or from front to. back. It is pos-
body z-y. sible, for example, to cut the eye on one side of the head and miss it altogether
on the other. The use of a ruler laid across a · total mount drawing will often
enable the student to appreciate the plane of sectioning. ·
Records. As the study progresses a record should be kept of the various structures encountered.
The most convenient method is to list the location of the structure as to slide, row, and section alongside ·
the name of the structure in the manual, thus:
1 I 5 I 17: Thyroid gland.
Since most structures persist through many sections, it is not necessary to state the exact section in which
they occur but only the slide and row.
It is also desirable to make small sketches of the structures identified as well as their immediate
surroundings. Tl.!ese drawings are not intended t~ be elaborate but should be placed in the manual
in the space provfded wherever an asterisk occurs.
EMBRYONIC MEMBRANES
In beginning the study of a set of sections of the 48-hour chick, the amnion, chorion, and yolk sac
show to advantage in the first few sections through the head. These structures may always be recog-
nized by the following characteristics.
*Amnion. A thin membrane composed of ectoderm (inner layer)
and somatic mesoderm (outer layer) lying next to the body but
separated from it by a space, the amniotic cavity.
~ .Chorion. A thin membrane also composed of ectoderm (outer
layer) and somatic mesoderm (inner layer) forming a second
covering layer outside the amnion. The space between amnion
and chorion is the exocoel.
: : :; * Yolk Sac. A thin membrane composed of endoderm and splanch-
nic mesoderm lying outside the amnion. It may be distin- I
guished from the chorion by the fact that- its mesodermal layer ·
contains vitelline blood vessels, whereas none are present in the
chorion.
 48-HOUR EMBRYO 61

ECTODERMAL DERIVATIVES

EPIDERMIS. A single layer of ectoderm cells forming the outer covering of the body.
BRAIN AND SPINAL CORD. In working down into the series the first prominent structure encountered is
the brain.1 Learn to recognize its parts by the following diagnostic features:
- M yelencephalon. Very thin roof. Associated with otic vesicle.
M etencephalon. Located in front of the myelencephalon but behind or posterior to the narrow
isthmus which separates mesencephalon and metencephalon.
M esencephalon. Located in front of the isthmus. In cross-section it is thick-walled and almost
always round.
. Diencephalon. Associated with the eye.
Telencephalon. Associated with the nose. Located at the anterior end of the neural tube.
Spinal Cord. The thick-walled, narrow, tubular extension of the neural tube posterior to the
myelencephalon.

2
ORGANS OF SPECIAL SENSE

-> * Otic Vesicle. Situated alongside the myelencephalon. Has the

form of a thick-walled pit of ectoderm widely open to the out-
,11 side of the head .
• •
I ~ Jf·~ -

- * Optic Cup and Lens. Situated lateral to the diencephalon. The

developing eye is made up of two primordia, as follows:
Optic Cup. A double-walled, cup-shaped structure attached
to the diencephalon by the narrow optic stalk.
Lens. A thick-walled, pitlike invagination of surface ectoderm
which will ultimately occupy the mouth of the optic cup.

*Olfactory (Nasal) Placode. Situated on the ventrolateral sur-

faces of the head and associated with the telencephalon. A
.v. thickened plate of surface ectoderm just beginning to invag-
inate.

CRANIAL NERVES. At this stage of development certain of the cranial nerves have already appeared,
namely: V, VIII, and IX. Each of these possesses a ganglion, which as shown in Fig. 23, has the
appearance of a heavy ectodermal cellular condensation in the mesenchyme s!tuated alongside the
1 It is well to remember at this point that as you get deeper into the series the front end of the neural tube will eventually

separate from the hind end, owing to the flexing of the head. This also applies to other structures such as the notochord and
the digestive tube. Hence it may be necessary to follow certain structures into the front end of the head first and then repeat
the study, tracing them posteriorly. .
2 The organs of special sense follow a general plan in early development. They are first set aside as thickened ectodermal

plates or placodes which undergo invagination. In the case of the optic cup, however, the process is complicated by the fact
that the sense placode is first taken in with the invaginating brain and is later evaginated as the <?J>tic vesicle, the outer surface
of which is later invaginated again to form the optic cup.
52 ANATOMY OF CHICK EMBRYOS

brain. Using the otic vesicle as a landmark, identify the following nerves associated with the myelen-
cephalon:
* A. Anterior to otic vesicle.
(VIII) Acoustic ganglion of the auditory nerve. Located im-
c mediately in front of, or anterior to, the otic vesicle and also
slightly ventral to it.
(V) Semflunar ganglion of the trigeminal nerve. This ganglion
is quite large and lies in front of VIII near the anterior end
/. of the myelencephalon.
* B. Posterior to otic vesicle
(IX) Superius ganglion of the glossopharyngeal nerve. A heavy
mass of nerve cells lying immediately posterior to the otic
• :vesicle and slightly dorsal to it. If present, this ganglion is
usually quite small.
I 2
ENDODERMAL DERIVATIVES AND ASSOCIATED STRUCTURES

A. MOUTH, PHARYNX, AND ASSOCIATED STRUCTURES (FIG. 29).

In its original form the fore-gut extended into the head fold of the 24-hour chick as a flattened
tubular structure ending blindly just behind the forebrain. At this stage the anterior end has acquired
an opening to the outside, the mouth. Th~ is developed by .the junction of the fore-gut and stomodeum.

....,,.,:,,/;;7· ,.... \)i!,,·'%Lf' Rathke's

pocket
Gill clefts .,·.~
1-111 r~
/~::
' lnfundibulum

- - Diencephalon

FIG. 29. Diagrammatic section of the head of the 4S-hour chick showing relation of the stomodeum to the developing mouth and
pituitary body. '

The latter is formed as an invagination of ectoderm meeting the ventral side of the fore-gut but not
quite ·at the end. Thus a short, blind p cket of the fore-gut protrudes anteriorly as the preoral gut or
Sessell's pocket. The closing plate of ectoderm and endoderm formed by the meeting of the stomo-
deum and fore-gut soon ruptures, thus producing the oral cavity. The mouth, therefore, is lined partly
with ectoderm and partly with endoderm.
,r 4
·,., * Hypophysis. From the stomodeal ortion of the mouth cavity
a_flattened tubular evagination.....arises o!.!_the dorsal surface and
grows upward to make contact with a similar evagination from
thefloor of the brain. These structures are known as Rathk_!l's
pocket and the infundibulum, respectively. They eyentua]ly
join and form the beginnings of the hypophysis ·or pituitary
d1~ pody; .
Visceral Clefts. Just posterior to the mouth a series of paired visceral clefts are developed on the
side .walls of the fore-gut in the region known as the pharynx. Each of these vis<l_eral clefts is produced
by the outward evagination of an endodermal visceral pouch from the side wall of the gut to meet a.Ii

..
 48-HOUR EMBRYO 53

inward invagination or branchial groove formed from surface ectoderm. The process is similar to that
by which the mouth is formed. The rupture of the ectendodermal closing plate produces a visceral
cleft or gill slit.1 At 48 hours three pairs of these are present.
:] Thyroid Gland. In the floor of the pharynx opposite the second
branchial arch the thyroid gland has begun to form as a rounded
ventral diverticulum which communicates with the pharynx by
I
- :q --- 0
means of the widely open thyroglossal duct. The cells consti-
tuting the wall of the thyroid diverticulum are characteristic-
ally tall and columnar at this stage.
B. DIGESTIVE TUBE POSTERIOR TO PHARYNX
Fore-gut. A thick-walled tube of small bore posterior to the
pharynx. Circular in cross-section. ,,
•Liver. Just in front of the anterior intestinal portal there is a
narrow ventral diverticulum from the floor of the gut. This
projects downward toward the meatus venosus and by cellular
I proliferation invades the walls of the vitelline veins which are
c
~c·~; '• prominent at this level, thus producing irregular masses of cells
which represent the primordium of the liver.
•Anterior Intestinal Portal. Posterior to the liver diverticulum
0 the ut ·continues as a tube for a short distance and then its
I
I
floot o~ns out ventrally. This marks the anterior intestinal
I portal.
... ' Mid-gut. From the anterior intestinal portal the gut continues
backward as the floorless mid-gut until it reaches the posterior
intestinal portal which marks t e gmning of the tail fold.
~t. At the level of the _posterior intestinal portal the gut
again assumes a flattened tubular appearance and extends back

• •
MESODERMAL DERIVATIVES
-
into the tail' fold as a blind tube, the hind-gut.

_,;, * Mesenchyme. Loose reticular embryonic connective tissue. Con-

stitutes the general padding mafenal between organs and sur-
face ectoderm.
- -- > • Notochord. CylindricalL rodlike (sometimes sinuous in outline).
Situated directly beneath neural tube. Represents the embry-
onic forerunner of the skeletal system and hence curves to
follow the contour of the floor of the brain.
CffiCULATORY SYSTEM

CARDINAL VEINS. Have the__g@eral form of the' letter H, of which: the u er limb represents the
anterior cardinals, the lower limb represents the posterior cardinals, and the cross barrepresents
the common cardinals.
Anterior Cardinals. Drain the head. Arise from a capillary J?lexus over .the_brain. The main
trunks lie at t e ventrolateral margins of the neural tube. They follow the neural tube closely
until the level of the heart is reached_ At this level they work ventrally but lateral to the
dorsal aorta to join the common cardinals. ·
1
It should be noted at this point that the visceral clefts are separated by pillars of tissue, whicli are known as visceral or
branchial arches. These are important at this stage as the territory through which certain blood vessels, the aortic arches, flow.
They are identified numerically as follows: visceral arch I lies in front of visceral cleft I: visceral arch II lies in front of
visceral cleft II, etc. (See Table 1 on pagl'l 7, Figs. 10 and 31.)
54 ANATOMY OF CHICK EMBRYOS

Common Cardinals. Receive blood from both anterior and posterior cardinals. Drain into the
dorsolateral angles of the sinus venosus.
Posterior Cardinals. Drain the posterior end of the body. Situated dorsal to the mesonephric
tubules. Join the common cardinal .veins.
VITELLINE VEINS. Return blood to the heart from the yolk sac.
--
B. ARTERIAL SYSTEM
AORTIC ARCHES AND RELATED VESSELS. In a section such as that shown in Fig. 24 we encounter the
upper end of the first aortic arch near the point where it joins the dorsal aorta and where the
latter is prolonged forward into the head as the internal carotid artery. Trace these vessels
deeper into the series and also identify the second and third aortic arches, etc., as follows:
,f
*Internal Carotid. Runs forward into the head to form a capillary
plexus closel_y_ associated with the brain.
* First Aortic Arch. Separates from the dorsal aorta. Runs through
the first branchial arch. Eventually swings into the mid-line
at its point of origin from the heart.
Dorsal Aortae. Situated beneath notochord, lateral to the mid-
re line and iust dorsal to the gut. They receive, in order pos-
teriorly1 the first, second, and third aortic arches. At the level
of the heart ihey fuse into a single vessel. Finally divide again
to form the two caudal arteries.
* Second Aortic A.,rch. Runs through· second visceral arch lateral
op arynx. Note association with thyroid _gland.
ird Aortic Arch. Runs through third visceral arch and hence
is found a few sections posterior to the second arch.
V itelline Arteries. Arise from dorsal aorta just posterior to level
atwhich the vessel splits to form the caudal arteries.
Intersegmen'tal Arteries and Veins. Small paired vessels which
supply the somites. Situated between somites. Arteries arise
from the dorsal aorta. Veins join the anterior or posterior
cardinal veins, depending on the level o( section.

* C. PARTS OF HEART
Bulbus Arteriosus. Usually circular in cross-section. Its anterior
end lies in t e mid-line beneath the aortic arches. Its posterior
nd swings laterally to the right and so comes to lie near the
chorion.
V:entricle. Usually oval in section. Its long axis lies across the
section. . It is farthest out ventrally from the body proper. It
has a delicate endothelial lining. Its wall is thickened by the
presence of a reticulum of developing muscle cells.
Atrium. Thin-walled. Situated ventral to sinus venosus.
Sinus Venosus. Thin-walled. Situated in mid-line bflneath gut.
Receives the common cardinal veins at its dorsolateral angles.
M eatus Venosus. Althou_gh not a part of the heart, it is. .a. 4.!gle
u e entering the sinus venosus. It represents the fused vitelline
vems.
 48-HOUR EMBRYO 55
DIFFERENTIATION OF THE SOMITES
It is usual to consider the lateral sheet of mesoderm as being made up of three regions which are
clearly distinguishable at this stage (Fig. 30). ·
(a) Somite proper. The :rnesoderm _lying alongside the notochord is divided by transverse constric-
tions into small paired blocks or somites from very early stages of development. The cavity in the
somi_te_is the myocoel. Each somite undergoes a. typical differentiation, which is clearly shown in any
of them, occurring about mid-way back in the body. It will be seen by reference to Fig. 30 that three
regions can be identified. Of these the ~ii!g layfil: lying just below the surface ectoderm is well
marked out and forms the dermatome, The hook of tissue continuous with the dermatome and adja-

Flo. 30. Differentiation of a typical somite as illustratf d by a section through mid-body of a 48-hour chicle.

cent to the sp~ord is. not very clearly marked out but in later develoE!?ent _giyes. rise. to skeletal
~ence is .known. as the ~ The remainder of the mesodermal block made up of the
bulk of the somite is the sclerotom it will give rise to skeletal structures.
(b) Nephrotome . ..J£. er portion of mesoderm lateral to the somite but not blocked off (in
the chick) by the transverse constrictions which separate the somites. The cavity in the nephrotome
is the nephrocoel.
( c) Lateral mesoderm. The broad lateral sheet of mesoderm not marked off by transverse divi-
sions. It is split tangentially into two layers, the somatic, or outer layer lying next to the ectoderm,
and the splanchnic, or inner layer, lying next to the e,ndoderm. The cavity between these layers is the
coelom.

COELOM
At this stag~ littl rogress has been made toward closing in the ventral side of the-body by lat-
eral undercutting. . Consequently the coelom is in wide-o_pen communication with the exocoel. There
is, ho~er, a blind pouchlike -extension of the coelom forward but lateral to the fore-gut in the region
of the heart (Fig. 25). The pericardiftl. cavity is continuous with the exocoel.

EXCRETORY SYSTEM
* PRONEPHROS. The excretory system of the chick is first developed as a pronephros or head kidney.
This sfructur~ is co.mp d of a_ll_erie of.proneR.hric. tubules which
grow backward and u~ite so as to form the prQnep]lric duct: At
48 hours a number of these tubules usually persist about the level
~. . of the fifteenth somite. Anterior to this they are degenerate.
Identify where possible:
Pro!';..ephric Tubules. Contorted thick-walled tu!?ttl_es. Equipped
with a e lifostome or opening to the coelom.
(
Pronephric Duct. Thick-walled, circular in cross-section. Formed
by union of backward extensions of pronephric tubules.
...
 56 ANATOMY OF CHICK EMBRYOS

* MESONEPHROS. The excretory system is represented in the 48-hour chick in the main by the mes- ·
onep os a series of individual mesonephric tubules which deve op
from the nephrotomes pos rior to the pronephros and communi-
cate with the mesonephric duct. In the ca.Se of the chick the neph-
rotome 1s tucked up under the spmite proper, and hence the mesone-
phric tubules appear to lie at the lateral angles of the somite. A
-- 'i;~~· close association with the dorsal aorta and posterior cardinal vefu
-. is ~vident. Identify:
" - - .7- M esonephric Tubule. Thick-walled. Contorted and irregular ·
in snape.
M esonephric Duct. Thick-walled. Circular in · cross-section.
Lateral to the mesonephric tubules which empty into the duct.
Represents a backward prolongation of the pronephric duct.

72-HOUR E MBRYO

GENERAL BoDY FORM

The 72-hour chick embryo is much larger than the 48-hour embryo, but the curvature of the body
is so great, owing to the presence of cranial, cervical, and caudal flexures, that its total length is only
about 7 mm. In general, the body may be divided into head, trunk, and tail regions, and, in addition,
the beginnings of the appendages are apparent in the form of anterior limb buds at the level of the
seventeenth to nineteenth somites and hind limb buds at the level of the twenty-sixth to thirty-
second somiteS;
The head, which is almost bent backward, shows a series of prominent bulges corresponding to the
divisions of the brain. The cerebral hemispheres of the telencephalon cause prominent lateral bulges
in the extreme front end. Diencephalon and mesencephalon are c.learly marked out, and, posterior to
the isthmus, the metencephalon and thin-roofed myelencephalon. Associated with the extreme ventro-
lateral surfaces of the head are the groovelike nasal pits. The optic cup and lens are clearly apparent
alongside the diencephalon, and the otic vesicle can be seen as a small saclike structure alongside the
myelencephalon. ·
The pharyngeal region is marked by the presence of the first three visceral clefts. The fourth,
although indicated, has not broken through. Anterior to the first visceral cleft the first visceral arch
is prominently developed, and the maxillary process, primordium of the upper jaw, has grown forward
to become attached to the head proper. The mandibular process, which is the primordium of the lower
j constitutes tlie main body of the arch. In total mount preparations the maxillary and mandibular
p ocesses have the appearance of a heavy, bilobed tissue mass lying just above the heart.
The heart is attached prominentl3{ to the ventral side of the body just below the pharynx. It is
still a looped structure (Fig. 22), and consequently the bulbus arteriosus as well as the more pos- ·
teriorly located ventricle form the uppermost limb of the heart. It will be noted that the anterior end
of the bulbus swings to the mid-line ventral to the gill slits. The atrial region is beneath the ven-
tricular limb but situated well forward. It communicates with the sinus venosus, which is located in
" the median line posterior to the atrium. The vitelline veins from the yolk sac converge toward the
posterior end of the sinus venosus.
In the body proper the neural tube swings backward in the mid-line and paired somites lie along-
side it. The somites, of which there are thirty-five pairs, begin just posterior to the otic vesicle and
extend back into the tail. The vitelline arteries leave the dorsal aorta at approximately the mid-level
of the body.
The tail fold is lifted above the blastoderm and curves slightly forward. At this level the allantois
protrudes out ventrally as a prominent vesicle.
Covering the whole body are the delicate amnion and chorion, which are almost entirely closed in
by fusion of the head and tail folds.
 /
'7 t.,
/

72-HOUR EMBRYO 57
Laboratory Directions
Study carefully a total mount of a 72-hour embryo, and make a drawing showing the structures
identified.

~ /1101· /'1, J,l,., ,.

Pr~ " ,~ al I
-)-Je;a,+ .

' Cj ,. ,,,,

> .J 'J

A ,,, , s

.·
.,_,,

72-Hour Chick Embryo.

58 ANATOMY OF CHICK EMBRYOS

DIBECTIONS FOR STUDY OF TRANSVERSE SERIAL SECTIONS

For convenience in working out and identifying the structures of the 72-hour chick we shall study
ectodermal, endodermal, and mesodermal derivatives as individual units after first considering the em-
bryonic membranes. A record slio.uld~e made · . · · in the form
of dra;vi_ngs of the various structu~s~if'cfthe listing of their location in the series.

EMBRYONIC MEMBRANES
Just as in the 48-hour chick series, the embryonic membranes are prominent and easy to identify
in the first few sections through the head as follows:
*Amnion. Lying next the embryo and forming a complete sac
· around the head. Composed of an inner layer of ectoderm and
~,.....,- an outer layer of somatic mesoderm. .·
---:---..~ ~ *Chorion. Lying outside the amnion and forming a single sheet.
Composed of an outer layer of ectoderm and an inner layer of
somatic mesoderm.
Yolk Sac. Lying outside the amnion and forming a single sheet.
Composed of endoderm and splanchnic mesoderm. Contains
vitelline blooq vessels in the mesodermal layer.

ECTODERMAL DERIVATIVES
....
EPIDERMIS. The outer covering of the body composed of a single layer of flattened cells.
BRAIN
M yelencephalon. Thin-roofed. Associated with otic vesicles. Side walls marked by a series of in-
dentations, the neuromeres.
Metencephalon. Anterior to myelencephalon and not clearly marked off from it. Thick-walled and
tapering inward toward isthmus.
Isthmus. Constriction between metencephalon and mesencephalon.
M esencephalon. Anterior to metencephalon. Characteristically thick-walled and almost round m
cross-section.
Diencephalon. Associated with eye.
i6'! * Epiphysis. A small but prominent rounded bulge on the dorsal
side of the diencephalon. Situated in the mid-line.

* Infundibulum. A flattened diverticulum arising from the floor or

ventral side of the diencephalon and directed toward the stomo-
deum.

*Cerebral hemis heretJ. Prominent rounded lateral bulges of the

;;:;..>---=-----=---'"-
y
telencephalon.

I~ 1 '1· ~o

Telencephalon. Associated with the nasal pits.

 72-HOUR EMBRYO 59
SPINAL CORD. The extension of the neural tube posterior to the myelencephalon. A narrow thick-walled
tube. Situated just beneath the dorsal surface of the body.
ORGANS OF SPECIAL SENSE
* Otic Vesicle. Thick-walled, ho!low oval vesicle. Lateral to myel-
encephalon. Frequently retains connection to the outside of
the head by the endolymphatic duct.
q.H: ! Optic Cup and Lens. The developing eye is made up of two
j c oo•C- primordia situated lateral to the diencephalon, as follows:
..__.~""t Optic Cup. A double-walled, cup-shaped structure. Attached
.;,; to .dience halon by the optic stalk on the under side of which
is a groove, the choroid groove. Its thick inner layer is the
'P.g .... ,,4., /. ~ .. ~tinal layer. Its thin, dark, outer layer constitutes the pig-
'Ir!! n il ' a.., r
mented layer.
,,,.. Lens. A rounded, ball-like structure occupying the mouth of
the optic cup.
*Olfactory Pit. · Situated on the extreme ventrolateral surfaces of
~1. • .i- the head adjacent to the telencephalon. Thick-walled. Ex-
ternal orifice much more constricted than at 48 hours.

CRANIAL NERVES

NERVES ASSOCIATED WITH MYELENCEPHALON

* A. Anterior to otic vesicle and listed in order anteriorly
(VI II) Acoustic ganglion. of the auditory nerve.
(VII) Geniculate ganglion of the facial nerve. Closely associated
with VIII.
(V) Semilunar ganglion of the trigeminal nerve. (Becomes tri-
partite.)

* B. Posterior to otic vesicle and list~ in order posteriorly

(I'Jr) Superior ganglion of the glossopharyngeal nerve.
(X) Jugul_f. ganglion of the vagus nerve. (Seldom present.)
(XI) Spinal accessory. The external branch is best developed at
this stage and appears as a long band of fibers adjacent to the
myelencephalon.
0 (XII) Hypoglossal nerve. When present the roots of this nerve
appear as a row of small dots ventrolateral to the floor of the
. .. . brain at the level where it is just separating from the spinal cord.

I. '/
 60 ANATOMY OF CHICK E~RYOS

NERVES ASSOCIATED WITH MESENCEPHALON

Bff IS *(III) Oculomotor. This nerve arises from the floor of the mesen-
cephalon. Hence it is found beneath the brain and its basal
portion is directed almost straight a.way from the bra.ill.

NERVES ASSOCIATED WITH TELENCEPHALON

E i lS/j . *(I) Olfactory. This nerve runs laterally from the wall of the tel-
/ 3 /(u~ I 'J>,+. encephalon to the nasal pit.
1 0 .(., ..,
I))'

'
: .

*SPINAL NERVES. The primordia of the segmentally arranged dorsal root ganglia are loc'a ted dorsolateral
to the neural tube as paired condensations of neural crest tissue.
They are usually somewhat difficult to recognize (Fig. 30).

81 S/
ENDODERMAL DERIVATIVES AND ASSOCIATED STRUCTURES

Oral Aperture. The external opening of the mouth cavity formed by invagination of the ectodermal sto-
modeum. Situated beneath or ventral to head. Bounded by:
Maxillary process of first visceral arch which has grown forward and become prominently attached
to the head proper.
Mandibular process of first visceral arch which constitutes the main body of the arch.
Mouth Cavity; The anterior chamber of the digestive tube. Originates from stomodeum and fore-gut.
*Pituitary Body; Consists of two closely associated ectodermal primordia:
(1) Rathke's pocket, a dorsal diverticulum of the stomodeum.
(2) Infundibulum, -a ventral diverticulum of the diencephalon.

Pharynx. The broad anterior end of the original fore-gut posterior to mouth cavity. Flattened dorso-
ventrally. Marked by presence of visceral clefts.
 72-HOUR EMBRYO 61
Visceral Clefts. Paired lateral openings extending from the sides of the head into the pharynx. Formed
by the junction of:
(1) Visceral pouches, which represent lateral outpocketings of the gut.
(2) Visceral grooves, which represent inpocketings of the surface ectoderm. Three pairs are present
-a fourth usually so.
4t _this time it is highly desirable to learn to identify the visceral grooves and the visceral arches
which lie between them. This is i!nportant because of their relation to the aortic arches. ~ page-~
In such a section as that in Fig. 3!,,,_the arches and grooves are clearly indicated. Learn to identify:
·<.:_/le/A~ •s;/t?//8 •t:!074J ~N/n M.14.J

Fla. 31. Section through the head of a 72-hour chick shoWing relation between visceral arches and groove&
B.//.:r/ :i ~ ~
(la) Maxillary process of arch I attached to the Head proper.
(lb) Mandibular process of arch I attached to the body proper.
(1) Hyomandibular cleft between arches I and II.
(II) Hyoid arch next dorsal to lb.
(2) Visceral cleft II between arches II and III.
(III) The third arch.
(3) Visceral cleft III between arches III and IV.
(IV) The fourth arch.
* Thyroid Gland. A rounded median ventral diverticulum from the floor of the pharynx.

1-_:e
11.,,,,..
J
, i. ... Situated opposite the second arch.

* Laryngotracheal Groove. Posterior to the fourth pharyngeal pouch the gut elongates ventrally. The
dorsal portion represents the oesophagus, the ventral, the begin-
d !f ~ :i. r nings of the trachea.
" I

B~,. ,, , h,
Ti c. ea b - - -

,, -.,.,
62 ANATOMY OF CHICK EMBRYOS

Oesophagus. The oesophagus continues beyond this level as a thick-walled oval tube.
..* Primary Bronchi. The posterior end of the tracheal portion of the laryiigotracheal groove bifurcates
~ /'I into two lateral budlike prominences, the primary bronchi. Note
Oe.t>•r
the close a.Ssociation of the pulmonary vein with the bronchi. This
vessel flows immediately into the left side of the atrium of the heart.·

--

81 $
e.1

Stomach and Duodenum. These follow the oesophagus but as yet they are not clearly defined except
for the fact that the posterior boundary of the duodenum is marked by the development of the liver
diverticulum.
*Liver. A ventral diverticulum from the gut. Its tissues spread laterally and invade the walls of the
.....
vitelline veins ~d meatus venosus.

Ant:ri;r ·1~t:stinalPortal. Just posterior to the level at which the liver arises the gut elongates ven-
trally once more to open out on the yolk sac. This marks the anterior intestinal portal.
*Pancreas. At the level at which the ventral elongation of the gut begins, a thickening of the dorso-
lateral wall of the gut on the right side can be seen. This is the
beginning of the pancreatic diverticulum.

Mid-Gut. Posterior to the anterior intestinal portal.·' Has no floor. Lateral walls thick. Bounded an-
teriorly by the anterior intestinal portal and posteriorly by the posterior intestinal portal.
Hind-Gut. Traced posteriorly the mid-gut closes in again on the ventral side at the level of the posterior
intestinal portal, thus forming a tube extending into the tail fold.
Cloaca. The gut becomes narrowed from side to side at the level of the future cloaca. Receives the
mesonephric ducts at its dorsolateral angles.
 ' 72-HOUR EMBRYO 63

• Allantois. A small, rounded, bladderlike, ventral diverticulum from the hind-gut growing out into the

s, - ... -
Postcloacal Gut. The extension of the hind-gut into the tail fold posterior to the cloaca.
MESODERMAL DERIVATIVES
M esenchyme. Loose, reticular embryonic connective tissue filling spaces between organs in the head.
N otochord. A solid rodlike structure representing the embryonic forerunner of the skeletal system.
Located beneath the brain and spinal cord. Curves to follow contours of the floor of the brain. Cir-
cular in cross-section--sometimes elongate and sinuous.
CIRCULATORY SYSTEM
A. VENOUS SYSTEM
CARDINAL VEINS. These vessels still have the general form of the letter H but conform to the con-
tours "Of the body and are still paired throughout.
Anterior Cardinals. Drain the head.
Common Cardinals. Receive anterior and posterior cardinals. Drain into dorsolateral angles of
sinus venosus.
Posterior Cardinals. Drain posterior part of body.
Vitelline Veins. Large paired vessels which drain the yolk sac and converge to form the meatus
venosus before entering the sinus venosus.
Allantoic Veins. These vessels are not yet well developed but are represented by a capillary
plexus in the ventrolateral body wa.11 posterior to the level of the sinus venosus.
Pulmonary Veins. Closely associated with the primary bronchi. Located ventral to the bronchi.
Drain into the left side of the atrium.
B. ARTERIAL SYSTEM

... • AORTIC ARCHES AND RELATED VESSELS.

$~~m•I ... 'Po•-s-.I A•.J, Since the aortic arches fl.ow through the corresponding
visceral arches they can' be easily identified when once the visceral
_ arches have been ·worked out as described above (pages 7 and
61). It is well to begin the study of the aortic arches with a sec-
tion such as Fig. 31, working forward in the series to their union
with the radices of the dorsal aorta and then backward to their
drigin from the bulbus arteriosus. Remembering that the aortic
arches fl.ow through the corresp~nding visceral arches, identify
aortic arches II, III, IV, and V-VI (usually combined).
•Internal Carotid Arteries. Anterior extension of dorsal aortae to
form capillary plexus over brain.
Dorsal Aorta. Paired radiees dorsal to pharynx. These vessels may
be confused with the anterior cardinals at this level but can be
readily distinguished because of the fact that they are situated
more toward the mid-line. Traced posteriorly they fuse to form
a single vessel. ·
 ANATOMY OF CHICK EMBRYOS
64
i.1/t.,.,, -.1 c,,,.;, a
'[;, ., *External Carotid Arteries. Minute paired vessels arising from the
If T
> base of aortic arch II. Extending ventrolaterally into the man-
1;.
1 dibular process of arch I.

Pulmonary Arteries. Small paired vessels arising from aortic arch VI and directed posteriorly
toward the lung buds. :Di "
• I ntersegmental Arteries ( arnfV eim). Small paired vessels ramifying between the somites. First
encountered posterior to the otic vesicle. The arteries anastomose
with the intersegmental veins which in turn join the anterior or
posterior cardinals.
-.:r. e I

M esenteNc Arteries. The dorsal aorta. can be followed posteriorly for a. considerable distance,
giving off small mesenteric arteries ventrally to the gut and sm8.ll paired dorsolateral interseg-
mentals between the somites. ·
-; ' Vitelline Arteries. A pair of large vessels which are given off from the dorsal aorta ventrally,
and at about the level of somite twenty-two course out over the yolk sac.
Caudal Arteries. Extensions of dorsal aorta. into the tail fold. At the level of the posterior limb
buds these usually give off lateral capillaries, which are the forerunners of the iliac arteries,
and which in turn give off small vessels to the allantois, the allantoic arteries.
• c. PARTS OF THE HEART. The heart of the 72-hour chick is greatly increa~d in size but it still has
the general form of a looped tube. It is attached to the body proper
at two points: (1) In the region beneath the visceral clefts by the
bulbus arteriosus, and (2) at a more pos~rior level by the sinus
venosus. The looped tube .is appended ventrally. Hence, while it
is usually possible to find a section where the regions of the heart
can be easily identified, it is always necessary to trace the parts of
the loop very carefully.
Sinus Veno8U8. Attached to ventral side of body. Receives right
and left common cardinals at its dorsolateral angles. Receives
the meatus venosus representing the fused vitelline veins at
its posterior end. Shifts to the right side of heart.
Atrium. Ventral to the sinus venosus. Lies nearest the yolk sac.
It is usually delimited from the sinus venosus and ventricle by
slight constrictions.
Ventricle. Ventral to atrium. Represents the outer curved por-
tion of the loop and hence lies transversely across the section.
Bulbus Arteriosus. Lies lateral to the other parts of the heart
and nearest the chorion. Communicates with the ventricle and
.. also with the aortic arches. It usually ap~ars sharply defined
as .circular or oval in cross-sections.
 72-HOUR EMBRYO . . 65

DIFFERENTIATION OF THE SOMITES. The parts of a typical somite are essentially the same as those
already encountered in the 48-hour embryo, as follows:
* SOMITE PROPER. Lateral to neural tube and consisting of:
Dermatome. Thick, dorsolateral plate. Primordium of the dermis.
M yotome. The dorsal "hook" of mesoderm continuous with the
1
dermatome but nearest the neural tube. Primordium of the axial
musculature.
Sclerotome. The main body of the somite consisting of loose mes-
enchymal cells.
Nephrotome. Ventrolateral to the somite proper. Primordium of
the excretory tubules.
Lateral M esoderm. The broad sheets of the mesoderm extending
'1 out lat~rally from the body proper between ectoderm and endo-
derm. Somatic mesoderm adjacent to the ectoderm is separated
from splanchnic mesoderm by the coelom.
COELOM
At the 72-hour stage the lung buds have begun to develop. They extend out laterally into the an-
terior coelomic pouches which now become part of the pleural cavity. They are also being undercut by
medial extensions from this cavity so as to form the pleural grooves.
At the posterior end of the body the tail fold has developed and in it the cloaca is taking form. A
pair of backward extensions of the -<:oelom extend alongside the cloaca into the tail fold.
In the trunk the lateral undercutting is more extensive and consequently the coelom within the
' .
body is more clearly delimited from the exocoel.
The pericardia! cavit~is"* ~tgI conj~1!..mJ~.~ith the exocoel. ./. /-
EXCRETORY SYS'N;M ':J! 3 /I/ t>. !\ ~;).
/ * Mesonephros. This system of tubules already encountered in the
/ 48-hour 'embryo is still present at 72 hours. A close association
with the dorsal aorta and postcardinal vein is apparent.
~ M esonephric Tubules. Thick-walled. Contorted shapes.
":? M esonephric Duct. Thick-walled. Circular in cross-section.
Lateral to mesonephric tubule. Empties into cloaca.
* M etanephros. The mesonephros is repiaced in later development
by the metanephros, which is also made up of a series of tubules.
These, however, have a very different origin. The metanephros
is not always present at .n hours, but when beginning to develop
the following parts may be identified:
Ureter. A dorsal evagination from the mesonephric duct at
the point where it joins the cloaca. It may show several bi-
furcations.
N ephrogenous Tissue. A heavily staining, caplike condensa-
tion of nephrogenous mesoderm over the end of the ureter.
 PART Ill

ANATOMY OF THE 10-MM. PIG

EXTERNAL FORM

The 10-mm. pig is a much larger and more complicated embryo than the 72-hour chick last studied,
and it has not only begun to take on its final external form but it has also developed the primordia of
nearly all the organs and organ systems. The body is clearly divisible into head, trunk, and tail regions
and the limb buds are well developed .
.The head is bent sharply almost at right angles to the longitudinal axis of the body owing to the
cervical and cranial fl.exures. The head is marked by prominent lateral bulges corresponding to the lo-
cation of the five vesicles of the brain. At the ventrolateral angles of the head the olfactory pits are
clearly evident, bounded by the median and lateral na.Sal processes. The eye, lateral to the diencephalon,
bulges prominently. The depression surrounding 'it is joined to the nasal pits by the nasolacrymal
groove which marks the anterior boundary of the maxillary process of the first visceral arch. The man-
dibular process, or primordium of the lower jaw, which is the main body of the first arch, lies posterior
to the maxillary process, the primordium of the upper jaw. The mandibular process is separated from
the second or hyoid arch by the hyomandibular groove which gives rise to the external auditory meatus.
The tuberculated edges of the hyomandibular groove furnish small tissue masses which form the pinna
of the ear. The second arch and groove are followed by the third arch and groove, but the fourth and
succeeding arches are squeezed in beneath the head in the region of the future neck by the cervical
flexure and their location is indicated only by the cervical sinus, a pitlike depression on the side of
the head.
In the region of the trunk, the heart is visible through the thin epidermis just below the head. The
liver lobes and mesonephros lie posterior to the heart in the order given. The liver is approximately the
same size as the heart, but the mesonephros is very large, occupying most of the space between the an-
terior and posterior limb buds. Along the dorsolateral sides of the body adjacent to the spinal cord a
series of small bulges marks the position of the spinal nerves and indicates the number of somites present.
The anterior limb buds project from the sides of the body at the level of the heart and the posterior limb
buds at the level of the posterior end."of the mesonephros. The body stalk is attached to the ventral
abdominal body wall.
On the ventral side of the body at the base of the tail is a small rounded prominence, the genital
tubercle, which is the primordium of the external genitalia.

Laboratory Directions
Study a 10-mm. pig embryo, and make a drawing showing the external features. Label ·fully.

INTERNAL ANATOMY
General Directions
In studying the 10-mm. pig embryo it is advisable to work out ectodermal, endodermal, and meso-
dermal derivatives separately. A careful record should be kept as usual, and a aeries of small sketches
should be made recording the location and identification of various structures.
67
68 ANATOMY OF THE 10-MM. PIG

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10-mm. Pig Embryo.

 INTERNAL ANATOMY 69

EcTODERMAL DERIVATIVES

The brain, organs of special sense, cranial nerves, spinal nerves, and developing sympathetic ganglia
constitute the chief ectodermal derivatives which are present at this stage in addition to the epidermis.
These will be considered independently. Constant reference should be made to Fig. 32.
A. EPIDERMIS. A layer of cells constituting the outer covering of the entire body.

B. BRAIN .
1. VESICLES.The brain is already divided into five vesicles. These are encountered as you work
down through the series in the following order:
*(a) SECTIONS SUCH AS THOSE AT LEVEL 1, FIG. 32

Otic vesicle, ,.....~Isthmus

''

Fro. 32. Lateral view of dissected head of a 10-mm. pig showing relation of cranial
nerves to brain and organs of special eenBe.

Mesencephalon. Thick-walled. Section typically round in shape.

t sthmus. Constriction between mesencephalon and meten-
cephalon. .
..... M etencephalon. Just posterior to isthmus. Thick-walled. Widens
gradually behind isthmus. .
_..,M yelencephalon. . Posterior to metencephalon-tapering gradually
toward spinal cord. Roof of anterior part thin-walled. Side
walls thick and marked by a series of indentations, the neuro-
meres, which were once thought to represent primitive segmenta-
tion of the brain.
* (D) SECTIONS SUCH AS THOSE AT LEVEL 4, 32
FIG.
/f Diencephalon. Associated with optic cup. Section of the brain is
round at junction with mesencephalon. It gradually changes to
an oval at the level of the optic cups, which are connected to the
brain
--· _,,,,
by the optic
. stalk.
70 ANATOMY OF THE 10-MM. PIG

* ( C) SECTIONS SUCH AS OSE AT LEVEL 5, FIG. 32

Telencephalon. The telencephalon has already begun to develop
the two thick-walled cerebral hemispheres which bulge laterally.
Their cavities are the lateral ventricles .

.!.5/1 7

2. SAGITTAL SECTION. At ·this stage it is well to examine a single typical mid-sagittal section '
•
through the brain. In such a section identify:
Telencephalon. The most anterior vesicle of the brain which is
marked off from the diencephalon by two constrictions, namely
the optic recess in the floor and the velum transversum in the roof.
Diencephalon. Bounded anteriorly by the optic recess and velum
transversum an? posteriorly by the tuberculum posterius on the
floor of the brp.in.
M esencephalon. This part. of the brain is located in the region of
the cranial flexure. It appears posterior to the diencephalon as
a .sudden expansion and is bounded posteriorly by the narrow
isthmus.
M etencephalon. This division lies just posterior to the isthmus.
It is marked by a comparatively thick roof.
~yelencephalon. There is no sharp line of demarcation between
this part of the brain and the metencephalon. The thin roof,
however, is quite characteristic.

c. CRANIAL NERVES. Working down through the series identify the various cranial nerves, as follows:
1. NERVES ASSOCIATED WITH ISTHMUS. LEVEL 1, FIG. 32
•(IV) Trochlear. Very small, inconspicuous. Leaves the brain
on the upper side of the isthmus where the mesencephalon just
joins the metencephalon. This nerve cannot be traced very far
at this stage but it eventually innervates the superior oblique
muscle of the eye. ·
 INTERNAL ANATOMY 71

2. NERVES ASSOCIATED WITH LATERAL WALLS OF MYELENCEPHALON. LEVEL 2, FIG. 32

* (a) ANTERIOR TO THE OTIC VESICLE (LISTED IN ORDER ANTERIORLY)
:1.d .P/ 3 7i[: (VII I) Auditory Nerve: Acoustic Ganglion. Innervates the otic
vesicle.
(VI I ) Facial Nerve: Geniculate Ganglion. CloselJ"'associated with
VIII. The nerve is directed posterolaterally and innervates the
side of the head, its fibers passing into visceral arch II.
(V) Trigeminal: Semilunar Ganglion. Situated at the broad an-
terior end of the myelencephalon. The ganglion is half-moon
shaped. Traced deeper into the body, three main branches of
the nerve appear: ·
(a) Superficial Ophthalmic Ramus. Arises from anterior end of
the ganglionic mass and passes toward the eye.
(b) Maxillary Ramus. Arises from mid-portion of the ganglion
·and passes into the maxillary process of the first visceral arch.
( c) Mandibular Ramus. Arises from posterior end of the gan-
glion and passes into the mandibular process of the first
visceral arch.
* (b) POSTERIOR TO OTIC VESICLE (LISTED IN ORDER POSTERIORLY) 1
(IX) Glossophp,ryngeal: Superius Ganglion,· Petrosal Ganglion.(' .:z.... z..
Situated juit posterior to the otic vesicle. Its fibers are directed
deeper into the head to innervate the tongue and pharynx. The
superius ganglion is found near the point of origin. The petrosal
ganglion is located farther down the nerve. Ca-/~ -
(X) Vagus: Jugular Ganglion; Nodose Ganglion. Its fibers at first
· r-qJl,~o those of IX but may be -traced well down into the
( body, where they innervate the visceral organs. The jugular
c/
,_11 ganglion is located near the point of origin. The nodose is found
1( by tracing the nerve somewhat deeper into the series.
,.. (XI) Spinal Accessory: Froriep's Ganglion. This nerve arises from
a series of small roots, which, therefore, appear like a row of dots
alongside the posterior end of the myelencephalon. The ganglion

::
I is obscure. The internal branch of the nerve runs parallel to the
vagus down to the shoulder, whereas the external branch runs
alongside the hind brain and parallels the spinal cord. It may
--/ thus appear as a long band of fibers adjacent to the myelen-
cephalon, or, deeper in the series, as a minute fiber wedged in
between the dorsal roots of the spinal nerves and the spinal cord.
3. NERVES ASSOCIATED WITH THE FLOOR OF MYELENCEPHALON. LEVEL 3, FIG. 32. A section through a
level such as this will show the spinal cord, base of the floor of the myelencephalon and the
mesencephalon, or possibly the diencephalon.
*(VI) Abducens. This small nerve arises from the ventrolateral
.C > portion of the floor of the myelencephalon. I ts fibers are di- >
rected forward and outward toward the eye, where it will even-
tually innervate the external rectus muscle.
* (XII) Hypoglossal. Arises as a series of roots from the ventro-
lateral sides of the myelencephalon before the brain narrows to
the spinal cord. Appears as a row of dots bridging the gap be-
tween the spinal cord and base of myelencephalon. This nerve
innervates the tongue.
 72 ANATOMY OF THE 10-MM. PIG

4. NERVES ASSOCIATED WITH THE FLOOR OF MESENCEPHALON. LEVEL 2, FIG. 32

* (III) Oculomotor. The fibers 9f this nerve protruding posteri-
7 orly arise from the floor of the mesencephalon. It may be traced
some distance toward the eye, where it will eventually innervate
certain eye muscles, namely: inferior oblique, superior rectus, in-
ferior rectus, and internal rectus. ·
,
c,

5. NERVES ASSOCIATED WITH DIEN CEPHALON. LEVEL 4, FIG. 32

(II) Optic. This nerve runs from the retina along the choroid fis-
sure on the under side of the optic stalk. It can rarely be seen at
this stage.
6. NERVES AS$0CIATED WITH TELENCEPHALON. LEVEL 5, FIG. 32
* (I) Olfactory. This nerve runs from the olfactory pit to the tel-
encephalon. It can usually (though not always) be seen.
(la) Terminal. Not present at this stage.

h 7
D. ORGANS OF SPECIAL SENSE
* 1. OTIC VESICLE. Lateral to the myelencephalon. Thick-walled and elongate oval in shape. Slightly
constricted into dorsal utllicular portion and ventral saccular
portion.
* Endolymphatic Duct. Situated between the otic vesicle and the
myelencephalon. Thick-walled and almost circular in outline.
Joins the lower or saccular portion of the otic vesicle.
/'/J .1.,, ~erA~/011

~~ y
* 2. OPTIC CUP AND LENS. Located alongside the diencephalon to which the optic cup is attached by
means of the optic stalk. The lens is almost solid and located within
the rim of the optic cup. .
Retinal Layer of Optic Cup. The inner layer of sensory cells of the
optic cup.
Pigmented Layer of Optic Cup. The outer layer of cells of the
optic cup darkened by the deposit of pigment.

j
t ~I J/ ' #-
I
 INTERNAL ANATOMY 73

Located at the anterior ventrolateral surfaces of the head. Has the form of a
thick-walled pit extending inward from the surface of the head.

C.5> /£"
-.

* E. SPINAL CORD. FIG. 36. At almost any level past the head, preferably one about midway through the
series, typical sections of the spinal cord and associated spinal nerves
c.7- can be found. Identify:
: :. Spinal Cord. Located dorsally just beneath the epidermis. Some-
what oval in shape.
-Central Canal or Neurocoel. The cavity of the spinal cord.
Ependymal Layer. The inner epithelial lining of the central canal.
Note the abundant mitotic figures.
Mantle Layer. The intermediate layer composed of neuroblasts
and spongioblasts-the precursors of the neurons and neuroglia
cells.
- Marginal Laytr. The outer layer of w ite matter composed of the
processes of neurons and neuroglia cells.
...
* F. TYPICAL SPINAL NERVE. Paired. Located ventrolateral to the spinal cord.
.;7 Dorsal Root. The bundle of afferent nerve fibers joined to the dor-
solateral wall of the spinal cord.
/ ?I Dorsal Root Ganglion. A prominent mass of neurons located on
the dorsal root close to the point of origin from the spinal cord
/ and lying alongside the cord.
.:;, Ventral Root. The bundle of efferent nerve fibers joined to the ven-
trolateral wall of the spinal cord.
~ Spinal Nerve. The main trunk formed by junction of the dorsal
and ventral root. It gives off three branches or rami, as follows:
Dorsal Ramus. The branch arising from the main trunk and
ascending to the dorsolateral body wall to supply the skin
and muscles of the trunk.
,... Lateral Ramus. The branch directed laterally to the ventral
body wall.
_.. Communicating Ramus. The branch qirected ventrally and
medially toward the ganglia of the sympat~etic system.
* G. NERVE PLEXUS
Brachial Plexus. At the level of the anterior limb buds the lateral
7•1-/ • rami of the spinal nerves are interconnected, thereby forming the
brachia! plexus which innervates the limb bud.
Lumbosacral Plexus. The nerve plexus supplying the posterior
limb bud.
 74 ANATOMY OF THE 10-MM. PIG

• H. SYMPATHETIC GANGLIA. Paired ganglionic masses lying dorsolateral to the aorta. These are most 0

readily recognized at about the level of the anterior limb buds

where the dorsal aorta is still paired.

..•
..
ENDODERMAL DERIVATIVES AND ASSOCIATED STRUCTURES

The original gut or endodermal tube has already become differentiated into the main parts of the
digestive system as well as the primordia of the respiratory apparatus. Beginning at the anterior -end,
follow the digestive tube backward in the series, identifying the following:
Oral Aperture. The external opening of the mouth cavity which is bounded by the maxillary and man-
dibular process of the first visceral arch. The maxillary process appears as a paired lateral prorp.inence
attached to the head below the eye. The mandibular process constitutes a thick ridge just posterior
to the maxillary process.
Mouth Cavity. Formed by the junction of the stomodeum and anterior end of the fore-gut. Its pos-
terior boundaries are indefinite. •
•
• Hypophysis. The hypophysis or pituitary body is form~d from ectodermal tissue contributed by
~c Rathke's pocket, which is a dorsal evagination from the stomodeum
and the infundibulum.

J., -1 .... J :L ... .. ?--

* Tongu'<: A flattened thick prominence on tne floor of the mouth formed of tissues contributed by all
.1-o .! /i / / s ~ ~three germ layers.

/ $ ~ l /"f ,
1...· -; ... ... . "
sl '(/,.$C'° ya I " ~
A .. J,./o-.-.. ,'f -r;, J. c..
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.4 I'

Pharynx. Posten ~to the mouth cavity. Compressed dorsoventrally. Marked by the presence of four
paired, lateral viscer_a l pouches. These meet the corresponding visceral grooves but the closing plates
do not rupture to form. a row of clefts. The visceral arches are pillars of tissue between the pouches.
The parts of visceral arch I form the boundaries of the mouth opening, the maxillary process being the
primordium of the upper jaw and the mandibular process, the primordium of the lower jaw. Next in
order posteriorly are arches II (the hyoid), III, and IV. Note the siinilarity to what has already been
encountered in the 72-hour chick.
Auditory Tube. Proximal end of visceral pouch I.
.s * Cavum Tympani. Dilated terminal portion-of visceral pouch I .
 INTERNAL ANATOMY 75

•Parathyroid Glands. Located at the upper (or dorsal) ends of third and fourth visceral pouqhes.
These primordia have the appearance of irregular thickenings or
Jlr buddings of the endoderm running into the adjacent mesenchyme
J a. "' o - of arches III and IV.
-fl.,.~c' ~
c
s> (::>ti) ..
., 2.- I

( . Thymus Glands. Located at the lower or ventral ends of the third and fourth visceral pouches. Their
appearance is similar to that of the parathyroids.

Thyroid Gland. A ventral diverticulum arising from the floor of the pharynx at the level of the second
visceral popch. At this stage of development it has already sunk deeper into the mesoderm and will
be much more readily identified when the aortic arches are studied.

• Laryngotracheal Groove. Posterior to the fourth visceral pouch the tube narrows greatly and develops
I a deep longitudinal depression on its ventral surface, known as the
s I"• 1 lo•e

-
laryngotracheal groove. The groove is prolonged posteriorly so
that its distal end separates off from the main tube, thus forming the
"'• trachea and leaving the oesophagus in the position of the original
tube.
Trachea. Ventral to the oesophagus. Round in cross-section.
.5 ~ * Eparterial Bronchus. A single rounded, budlike protuberance
on the right side of the trachea.
l 3/' .I
* Primary Bronchi. The rounded bifurcated ends of the trachea.

Oesophagus. Round and thick-walled in section. Forms the connection between pharynx and stomach.
• Stomach. Elongate and thick-walled. Broader than oesophagus. Supported by the dorsal mesentery.
1"t=:... Directed ventrolaterally and toward the right.
 76 ANATOMY OF THE 10-MM. PIG

Duodenum. Round and thick-walled. Begins where stoma.ch narrows posteriorly. Ends at point· of
(i .:i. / y. origin of the bile. duct and pancreas.
•Bile Duct ( ductus choledochus). A ventral tubular canal joining
the distal end of the duodenum. Thick-walled and directed ven-
trally toward the gall bladder within the liver tissue. ·
.; Gall Bladder. The enlarged sa.ccular distal end of the ductus
choledochus.
jf_iver (Fig. 36). Both ductus choledochus a.nd gall bladder are
entirely surrounded by the liver tissue, which is almost spongy
in appearance owing to the extensive ramifications of the blood
vessels and capillaries throughout the entire structure. As a re-
- suit, liver tissue is endodermal in origin as regards the secretory
tissue but has also a great deal of tissue of mesodermal origin
whch forms the walls of the blood sinusoids:
• Pancreas. A dorsal diverticulum from the duodenum near the
level at which the bile duct enters. It forms an irregular mass
lying in the dorsal tnese_ntery distinguishable mostly on the basis
of its staining reaction. Slight indication of ,the ventral pan-
creatic diverticulum is sometimes found in the form of a minute,
budlike prominence just at the point of entrance of the bile duct
into the duodenum. It is directed dorsally and to the right.

Intestine Prop"er. A thick-walled tube of small bore continuing bac~ard frQm the duodenum. It is
supported by the dorsal mesentery and is accompanied by the vitelline arteries. and veins.
• 0testinal Loop. At the level of th~ body stalk the intestine swings ventra11y out into the body stalk
and then back into 'the body cavity again. TJle two limbs of this
~. 1 loop are in close proxUnity, and at_its distal eiiel the ·narrow yolk·
rs...
(!_ ,JI'"'4D ,...... )
a ..... C¥/1/
stalk, which communicates wi~)l the .yolk sac, 'is ~ttached.
.
Colbn. After re-entering -the body cavity the tube continues backward tow~d tlie cloaca.
Cloaca. An enlarged cavity into which the colon enters (Fig. 33).
. .
NOTE. This structure is of considerable importance and will be dealt with more fully when the· nephri!J system is
considered. }/es ~
Allantoic stalk. A ~ · y 9(}1lapsed duct opening into the ventral side of the .cloaca. Runs -through
th body stalk to communicate with the bla.dderlike allantois, which
, .., - is sually removed before sectioning. .

/.I J, ./)
c1. t!1

(• I ... (

--
 INTERNAL ANATOMY 77

MESODERMAL DERIVATIVES
MESENCHYME. The loose reticular tissue which fills spaces between developing organs throughout the
body of the embryo. This material is of great importance in that it constitutes the forerunner of the
various types of connective tissue. It is composed of the cell body containing a nucleus, and proto-
plasmic processes which extend from cell to cell forming a delicate reticulum, in the interstices of
which a more or less fluid matrix is developed.
NOTOCHORD. A cylindrical rodlike structure forming the skeletal axis of the embryo. Circular in cross-
section. Located ventral to the spinal cord. Extends from region of mesencephalon to tail.
SOMITES. In the 10-mm. pig embryo there are approximately forty-four somites. These have already
undergone the early differentiation corresponding to that studied in the chick, and consequently in a
typical somite occurring about midway through the body we may recognize:
Dermatome. A dense strip of cells situated lateral to the spinal cord just beneath. the ectodermal
epidermis.
Myotome. The. proximal hooklike end of the strip of cells constituting the dermatome but bent
under or ventral to it and hence located closer to the spinal cord.
Sclerotome. The main bulk of the somite consisting of more loosely arranged cells.
M yocoel. The cavity of the somite. Located beneath .or ventral to the dermatome and usually
obliterated.
Nephrotome. The intermediate mesoderm is already differentiated into the complicated meso-
nephros which will be considered under the eoxcretory system.
Lateral M esoderm. The lateral sheets of mesoderm are already differentiated into a somatic and
splanchflic layer. Identify:
Somatic Layer. In <;:lose association with the ectoderm of the outer body wall.
Splanchnic Layer. Covering. tlie mesonephroi and also forming the mes®tery supporting the
. gut and associated-structures.
cOELOM. In the 10-mm. pig the pleural cavities and pleural grooves are already present at the level of
the lung buds. .Ill the region of the cloaca the backward extensions of the coelom form a saddle-
shaped. pelvic recess beneath the urog.enital sinus of the cloaca.
Within t,h.e trunk a blind ·c0elomic pocket has been formed in the mesentery owing to the shifting
of the positionif the stomac!i. As a result the dorsal mesentery is wrapped around the surface of the
stomach, as it were, and so forms a blind pouch, the omental bursa. This communicates with the
· coelom by means of the epiploic foramen ( foramen of Winslow).
PARTITIONING OF COElLOM. The transverse partitioning of the coelom into thoracic,and qbdominal regions
has begun in the 10-mo:i~ pig with the development of the septum transversuni/wiiich forms a sort of
· semicircular shelf attad1ed to the ventral body wall situated between the heart and liver.
MESENTERIES. During early embryonis development the splanchnic mesoderm envelops the gut. As
a result the digestive tube is attached to the dorsal coelomic wall by the dorsal mesentery and to the
ventral wall by the ventral mesentery. Only the dorsal mesentery persists, however, and the usual re-
gions may be recognized:
M esogaster-supporting the stomach .
. M e;oduodenum-supporting the duodenum.
M esentery proper-supporting the main body of the gut.
M esocolOn-supporting the colon.
In addition the gastrohepatic omentum is a persisting portion of the ventral mesentery joining the gut
to the liver.
 78 ANATOMY OF THE 10-MM. PIG

EXCRETORY SYSTEM

PRONEPHROS. In the pig a transitory pronephros is found in the most anterior nephrotomes. It has
already degenerated and been_ superseded by the mesonephros at the 10-mm. stage.
* MES~NEPHROS. The ~esonephroi which constitute the functional kidney of the 10-mm. pig are very
conspicuous and almost disproportionately large in comparison with
the rest of the body. They are located posteri{ r to the liver and
develop in behind the splanchnic mesoderm; hence they are said to
be retroperitoneal in position. They are so large, however, that
they protrude down into the coelom. Well-mar~d -differentiation
of parts is apparent at this stage, but it should be remembered that
individual mesonephric tubules make up the entire structure. Se-
lect a mid-body section (Fig. 36) and identify:
*Bowman's Capsule. The enlarged, double-walled chamber situ-
II " ated at the terminal end of the tubule and containing a glo-
merulus. Usually located near the mid-line. These structures
are usually quite irregular in shape.
~ Glomerulus. The knot of capillaries contained within the inner
cup of the Bowman's capsule. Irregular in shape, compact in
• I cri appearance. - Located near the mid-line.
~· M esonephri~ Tubule. The tubules lead from Bo~man's capsule
to the mesonephric duct. The tubules are hollow, curving
\ tubes. Usually situated laterally.
* M esonephric Duct. The common collecting duct leading back-
ward to the cloaca. Located on the ventrolateral border of
the mesonephros. Thick-walled and prominent. Usually flat-
tened oval in cross-section.
* METANEPHROS. The metanephros has already begun to develop at this stage. It arises from two
C.ft
sources, as follows: The secretory portions of the tubules take their
V A 1la1, /,Js
origin in the non-segmented nephrogenous tissue posterior to the
mesonephros and the collecting portions and duct develop as evagi-
nations from the mesonephric ducts. They may be recognized in
the following manner after tracing the mesonephric ducts backward
Coe.lo,., to the cloaca. \..
tre1~;e. re s. ) * Ureter or M etanephric Duct. A small evagination from the meso-
nephric duct at its point of entrance into the cloaca. Heavy-
%sf ·lue~I '· f walled. Directed dorsally at first, then anteriorly. Often bi-

•
• furcated several times to give the primor.Q.ia of the collecting
1' tubules of the first and second order. Its cavity represents the
primary pelvis of the future kidney.
* Primordium of the Secretory Tubules. Heavy condensations of
,,,.,, J. • 7A. J- nephrogenous mesoderm forming a cap over the end of the
es••7 .-.,; e I ureter or the collecting tubes originating from it.
v
------::;;. t!.10 . ,. ..2.

c I/ t,,_ rs,,. ' r

 INTERNAL ANATOMY 79

DIFFERENTIATION OF CLOACA (FIG. 33)

The cloaca is a structure of considerable importance at this stage, and consequently it should be
studied with care. Two regions may be recognized as follows:

/ ::
J_ *Urogenital Sinus. The broad, saddle-shaped anterior portion of
~ the cloaca which receives the allantois, and, at its lateral mar-
/
1
· "'"'; r -
~ gins, the mesonephric and metanephric ducts.
NOTE. It is important not to confuse this portion of the cloaca with the pelvic
recess of the coelom which also extends underneath the saddle-shaped urogenital
" ..5
sinus. In cross-section, both are curved, arc-shaped cavities but the coelom
lies within the arc of the urogenital sinus.
~ Rectum. The posterior narrow portion of the cloaca receiving
the gut and postcloacal gut.
* Cloacal Plate. The closing plate formed by apposition of the
~ lateral walls of the terminal portion of the cloaca. . '
-"
yV- 1"' :}/'~

Flo. 33. Reconstruction of the cloaca and related structures in the 10-mm. pig. Mes.
duct, mesonephric duct; Met. duct, metanephric duct; Neph. tissue, nephrogenous
tll.true; P.C.G., post cloaca! gut; Rect., rec:t\lDl; U.G.S., urogenital sinus.

REPRODUCTIVE SYSTEM

The gonads of the embryo are reptesented in the 10-mm. pig only by the merest rudiments in the
form of thickened genital ridges on the ventromesial side of the mesonephros. These structures ace
located about midway back along the mesonephroi. Identify:
1~ ~ * Germinal Epithelium. The irregular, thickened splanchnic meso-
/1 derm forming the surface layer.
• Primordial Germ Cells. Prominent cells with large vesicular
( nuclei situated in the tissue underlying the germinal epithelium.
Gonoducts. Since the sex of the individual pig is quite uncertain
/
at this stage, the gonoducts have not yet been differentiated.
'rot ) However, if the animal is to be a male, the mesonephric duct
hrr 1 will eventually function as a ductus deferens.
External Genitalia. No differentiation of the external genitalia.
can be recognized, but the genital tubercle is the primordium
of these structures. ..
80 ANATOMY OF THE 10-MM. PIG

CIRCULATORY SYSTEM

At this stage of development the circulatory system has become very complex, for the early stages
of development corresponding to those of the 33- to 72-hour chick have . already been completed. As
a consequence it will be found highly desirable to work out the circulatory system in separate units:

I. ARTERIAL SYSTEM AND HEART

A. ARTERIES OF THE HEAD

1. AORTIC ARCH SYSTEM

In the pig, just as in other vertebrates, the aortic arches develop as a series of six pairs of blood
vessels associated with the visceral arches. They also do not all develop at once and the ones formed
first soon begin to disappear. As a result, at the 10-mm. stage, aortic arches III, IV, and VI alone per-

-Radix aorta
-Nodose gang. X
. -Aortic arch Ill
Internal
Anterior- ·:~,¥5'"",.,;-~ "''1- carotid
cardinal vein ·~·~)liJ~d·~i~~~~~ artery

Fm. 34. Section through the head of a 10-mm. pig at the level of origin of the internal carotid arteries.

sist. These constitute the basis on which the adult plan of the circulation is established. Consequently
they should be carefully worked out in detail.
As a key point for tracing these blood vessels, select a section approximating level 3, Fig. 32, as
illustrated in Fig. 34. Such a section is significant because it shows the junction of the radix of the aorta,
the third aortic arch, and the internal carotid artery. The characteristic relation of these vessels is clearly
shown in the figure (subject to slight modifications, of course) as well as the typical association with ~
the nodose ganglion of the vagus nerve X. Trace out the courses of these vessels,_as follows: 1
SI INTERNAL ANATOMY 81

* (a.) Internal Carotid Artery. This vessel can be traced forward

in the head to the side of the diencephalon. In this region
it becomes broken up into a multiplicity of capillaries running
out over the brain. The main channel, however, can be
picked up again beneath the floor of the mesencephalon.
C ,,,. I Here the two vessels unite to form the basilar artery which
continues backward beneath the floor of the brain as a. some-
what uncertain channel to the beginning of the spinal cord.
At this level the basila:.r flows into the paired vertebrals which
continue backward, paired, to the level of the limb buds.
Here they anastomose with the subclavian arteries but also
continue backward and eventually fuse to form the spinal
artery which flows beneath the spinal cord.
* (b) Radix of Dorsal Aorta and Third Aortic Arch. These vessels
should be traced deeper into the animal. It will be found
that the paired radices maintain their original position in the
a 11...section relative to the spinal cord, etc., and that the paired
third aortic arch moves steadily a.way from the radix. The
two third arches eventually meet in the mid-line a.t a point
which represents their origin from the ventral aorta. Near

--
this level 1harp watch should be kept on the radix aorta for
the junction point of the fourth aortic arch.
NOTE. This is the point at which the developing *thyroid gland is readily iden-
....
tified as a small caplike condensation of endodermal tissue above the end of
the ventral aorta. i I •.-I-;
* ( c) External Carotid Artery. At the level where the two third
arches arise from the ventral aorta, the small external caro- 1
tids are found coursing obliquely· forw~d and outward into
the mandibular arch. · c,;~' ,. . . ... ~ '~, 1~
* (d) Fourth Aortic Arch. This vessel usually persists through
only a very few sections, and often its origin from the ven-
~ 7 tral aorta and junction with the dorsal aorta can be seen in
the same section. This gives it. a characteristic U sha .
* (e) Sixth Aortic Arch. A few sections posterior to the level at
which the fourth arch can be identified the ~·unct"-....
n point§)
c... -
of the sixth and dorsal aortae are encountered. en traced
deeper these vessels a.re seen to a.rise not directly from the
.. -· ventral aorta but rather from a separate trunk-the pul-

-
monary trunk-which has been separated off from the ven-
tral aorta. At this level it is usual to speak of the ventral
aorta as the systemic trunk. Note the characteristic Y shape
of the sixth arches at their point of separation from the_EUl.::.
~ryklJ.nk.

- (f) PUlmonary Arteries. These arteries which are quite small

arise from a point about midway along the sixth aortic arch
and may be traced deeper in the body toward the lung buds.
 82 ANATOMY OF THE 10-MM. PIG
'
2. HEART
* Bulbus Arteriosus. Follow the systemic and pulmonary trunks
deeper to their point of origin from the bulbus arteriosus. This
channel is very thick-walled and will be found to show a tend-
ency toward division into two channels near the ventric)llar
region of the heart. This is indicated by the presence of a ridge-
like thickening on both its dorsal and ventral wall. The ridge
j._,.J f is the beginning of the septum aorticopulmonale. The potential
S1"1t - t• ... l'.w'i.11tfulmonary trunk will be found to communicate with the right
,a., ,.iec. lat-
/)a/lie ventricle and the systemic trunk with the ~ft ventricle. -
The heart is already-divided into atrial and ventricular regions
and these in turn are incompletely partitioned into right and left
halves, thus forming the beginnings of the adult four-chambered
heart. As the bulbus arteriosus is traced backward in the series,
the parts of the heart become clearly evident in association with
s; 1 it, as follows:
''"/ Atria. Thin-walled and characterized by a number of saccular pro-
'!.,, .,."lf
jections on their upper lateral surfaces.
J.." f j Sinus Venosus. Associated with the dorsal surface of the right
,.. - atrium and communicating with the heart by an opening guarded
by a pair of thin valves which project into the atrium.
Interatrial Septum. The thin partition between the atria.
. /, /, * Poramen Ovale. The opening in the interatrial septum which al-
f!~f~ ..,, 11
~ 'f' lows communication between right and left atria.
_Atrioventricular Canals. The passageways forming a communica-
Cs 1 ~ tion between atrium and ventricle on each side. The tissues of
the rim of the right canal will form the tricuspid valve, those
of the left, the bicuspid valve. C. .r, ~ /-'/ - '1
Ventricles. Thick-walled and characterized by the presence of a
reticular syncytium of myoblasts in their walls. c
lnterventricular Septum. The thick partition bet~ the ventricles.
lnterventricular Foramen. The canal allowing communication be-
tween right and left ventricles. c
Endocardial Cushion. Thick masses of mesodermal tissue in the
dorsal and ventral wall of the ventricle of the heart at the level
of the atrioventricular canal. ,.. .s-, :a 1 - ?
 INTERNAL ANATOMY 83

3. ARTERIES OF THE BODY. As the dorsal aorta courses backward in the body, a series of branches
are given off, as follows:
* I ntersegmental .Arteries. Small paired vessels. Arise from.

-
the dorsolateral angles of the aorta. Run between the
somityrind to the neural tube.
* Subctfivian Arteries. Run out laterally into the anterior limb
buds where they spread out as a capillary network. "' ~
• Renal Arteries. Numerous small vessels arising laterally
from the aorta.. Supply the glomeruli of the mesonephros.
• Coeli.ac Artery. Arises from ventral side of aorta. at about
... / the level of the s~ach.
13 ISuperior MesenterjeArtery. Arises from ventral side of aorta
slightly post¢0r to the coeliac artery. Runs out along the
yolk stall¥
• Allantoiv or Umbilical Arteries. Lateral .vessels arising from
the aorta at the level of the posterior limb bud. Run out
along the allantoic stalk into the umbilical cord.
Iliac Arteries. Branch off the allantoic arteries. Run out
laterally into the posterior limb bud. C 1 2. 5'
Caudal Arteries. Backward extension of the dorsal aorta
• into the tail .
NOTE. It is desirable at this point to consider briefly the significance of the
status of the aortic arches .in the 10-mm. pig. The third aortic arches repre-
sent parts of the future internal carotid arteries. The dorsal aortae between
third and fourth arches are obviously dwindling in size and importance and
r "' r • will eventually be eliminated. Of the paired fourth arches, the left side alone
will persist while the right, along with the radix of the aorta on that side, will
disappear. The sixth arch on the right side will peraisi to the point of origin of
• the pulmonary artery, while the left will persist in its entirety until the time of
birth, as the ductus arteriosus, thus maintaining a sort of "by-pass" to the
dorsal aorta so that little blood actually goes to the lungs.

II. VENOUS SYSTEM

The venous system of the 10-mm. pig, like the arterial system, is already quite complicated, and it is
desirable, therefore, to work it out in a similar manner by studying individual units. In the pig, just as in
the chick, the cardinal system constitutes the basic series of veins draining the body proper in the early
embryo. Moreover, the vitelline veins bring back blood from the yolk sac, which, in the pig, is large and
of considerable importance as a nutritiQ11al organ. To these must be added the allantoic veins, which
are very large and prominent, since they constitute the drainage from the placenta. Finally a new
vessel, the post ca.val vein, is in process. of being established as one of the main drainage channels.

A. ANTERIOR CARDINAL VEINS AND TRIBUTARIES

The anterior cardinal veins are broad sinuslike vessels which course backward along the sides of
the brain lateral to the otic vesicle. At this level they are easily identified (Fig. 34). When traced
forward toward the forebrain they soon break up into a capillary network anastomosing with branches
of the internal carotid artery which cover the surface of the brain. These ramifications are difficult to
follow and give little information of importance. Traced posteriorly, however, they soon take up a
position dorsal and lateral to the paired dorsal aortae which they accompany posteriorly.
NOTE. (1) The typical close association of these vessels with the ganglia and fibers of the vagus nerve X.
(2) Intersegmental veins, draining independent areas of the side of the head, brain, and spinal
coi-d, can be seen at regular intervals emptying into the cardinals. -
 84 ANATOMY OF THE 10-MM. PIG

NoTE. (3) At the level of the visceral arches fairly large but uncertain vessels join the cardinals later-
ally and ventrally. These represent beginnings of the external jugulars.
( 4) At the level of the limb buds a number of vessels draining the appendages join the anterior
cardinals. From the most prominent of these the subclavian vein will develop.
B. COMMON CARDINAL VEINS
The anterior cardinals ultimately trace backward to joiri the
common cardinals which flow into the heart. The right common
cardinal will be seen to communicate directly with the right atrium
of the heart by way of the sinus venosus. Note the prominent
valve~ of the sinus venosus at the point of entry of the vein. The
left common cardinal behaves in a different manner. Instead of
flowing directly into the heart it will be found to narrow down
greatly, assume a close contact to the back wall of the left atrium,
and then loop across to the right side (still closely applied to the
heart), where it communicates ;vitJl the ,postc~v!l-1 ?"- ,._. "'
Relation of Anterior Cardinals and Common Cardinals to Precaval, Postcaval, and Coronary Veins.
The conditions encountered in the 10-mm. pig are highly significant, since they foreshadow the develop-
ment of the precaval and coronary veins. The precaval vein represents a shift in drainage of the venous
blood into the right side of the heart. Its main trunk is formed from the right common cardinal vein plus
a small portion of the proximal part of the right anterior cardinal. The tributary channels on both
sides are the sub_Q_lavian veins and the internal and external jugulars. Blood from the left side of the
Anterior
,rl External

I
cardinal jugular

Common-
cardinal
-- L. innominate
-----------: -~Subclavian

' ,. _. Coronary
Posterior
cardinal \
Precaval

R L R L
~ IG. 35. Changes occurring in the cardinal veins of the pig during development. Left, primitive condition; Right, adult condition .
Vessels added -------.

head ultimately will flow across to the right by the development of a new channel, the left innominate
vein, which will form an anastomosis from the left to the right (Fig. 35). As yet these developments
are not definitely under way. However, the striking manner in which the left common cardinal is
associated with ·the heart at this stage indicates that it is being taken over to drain the wall of the
heart, for, along with small portions of the proximal ends of the left anterior and posterior cardinals,
it thus becomes the coronary vein.
C. PULMONARY VEINS
At this stage the pulmonary veins are too uncertain and too poorly developed to trace successfully.
D. VEINS OF THE POSTERIOR PORTION OF THE BODY (FIG.36)
There are four main venous channels to consider in working out the drainage of the posterior part
of the body.
1. Posterior Cardinals. These are vessels of dwindling importance. They originally drain the
dorsolateral portion of the body wall and adjoining organs and hence are located dorsal to the meso-

,
 INTERNAL ANATOMY 85

nephros. Beginning at the point of junction with the common cardinal, they may be traced backward
deeper into the series and will be found to maintain their original dorsolateral position with reference
to the dorsal aorta. * As they are followed backward a level is finally reached where a n~w ~ of
tissue suddenly appears lying across the vessel, thus dividing it into an upper and lower half. The new .. ..._'rl-.~..__,
-.
$ 1/L - 11
If r 'II I

Subcardinal
vein

. .. 1J'w. 36. Section through mid-body of the 10-mm. pig•

tissue represents the anterior tip of the mesonephros, and the car·
dinal is thus separated into a dorsal posterior cardinal and a ventral
subcardinal, each of which receives numerous tributaries from the
mesonephros. The, posterior cardinals from this point back grad-
ually dwindle and may even be obliterated for a time. Farther back
in the body they may be picked up once more, again in a position
dorsal to the mesonephros, and from this point they may be traced
back into the tail.
2. H epati.c-Portal System. The hepatic-portal system is derived from the paired vitelline veins of
the younger embryo. These vessels originally course iµward along
the lateral walls of the gut from the yolk sac to the heart.
With the increase in size of the developing liver primordium,
which arises originally as a ventral diverticulum from the gut, the
closely associated vitelline vessels become broken up into a series of
capillary channels running through the liver. By the time the
10-mm. stage is reached the liver has become a large and prominent
organ; modification of the vitelline vessels is under way; and the
beginnings of the hepatic-portal system are apparent.
Posterior to the liver a series of transverse anastomoses, ta.
gether with the occlusion of intervening portions of the original
channels, transforms the vitelline veins into a vessel tending to
spiral around the intestine. It will be found that the right vitelline
forms the upper or anterior end of this spiral and that at this point
the vessel, now the' portal, suddenly moves across from the gut
into the liver. (Fig. 37).
86 ANATOMY OF THE 10-MM. PIG

Liver Ductus
venosus

Vitelline.-
veins
Umbilical
veins
Intestine - -
R L R L
Fla. 37. Changes occurring in the vitelline and umbilical veins of the pig during de-
. velopment. Left, primitive condition showing relation of ve1Eels to liver; Right,
condition at birth: vitelline veins give rise to the hepatic portal system, umbilicals,
to the ductus venosus which is occluded after birth. Vessels added -------, vessels
dropped ...•.•••

In studying out this system it is best to begin with a section taken through the level at which
this occurs. The condition is shown with almost diagrammatic exactness in such a section as Fig. 36.c?,L?fi_~
From this point the vessels should be traced forward and then backward in the series. The blood from
the portal is distributed by capillary sinusoids throughout the liver and is picked up again by the con- \..
vergence of another group of sinusoids in a main veSsel representing the upper end of the right vitelline
vein. This is the hepatic vein, and it usually joins the developing postcaval vein before entering the
heart. The proximal portion of the left vitelline vein contributes little save a few capillary sinusoids
in the liver. .
3. UmlJilical Veins (transitory). The blood distributed by the umbilical artePies to the allantois
is returned to the heart by the umbilical veins. These vessels course inward along the body stalk as a
pair of broad venous channels. At the level at which the umbilical cord joins the ventral surface of the,
body, they move into the adjacent liver and become involved with it as a series of capillary sinusoids
in a manner recalling the condition affecting the vitelline veins (Fig. 37).
c' .! 7 11. .,.j It is evident in the 10-mm. stage that the right umbilical vein
e.u.irr"fs losing its importance and dwindling in size. The..kf!J. on the other (! Yp/S
hand, becomes large and prominent and takes over a series of capil-
/.. lary sinusoids in the liver to make a direct path across the liver
,,e.,. from left to right toward the heart. This shortcut through the liver
.is..tbe-~ctus venosus. It usually joins the postcaval vein before
entering the heart.
In working out these vessels a section at a level approximating
Fig.· 36 should be chosen as a starting point and the umbilical
veins traced forward to the heart and back into the umbilical cord
" ~ where they are fused into a single vessel.
* 4. Postcaval Vein. A In the adult animal most of the blood is returned~ the heart by way of the
postcaval vein. This vessel has already begun to establish itself in the 10-mm. pig. The greatest im-
petus to its development seems to be the fact that blood from the mesonephros requires a more direct
passage to the heart than the circuit through the original posterior cardinals. As a result a pair of some-
what uncertain channels, ventral and medial to the mesonephros, first appear (Fig. 38). These are the
subcardinals. They receive tributaries from the mesonephros but empty into the upper ends of the pos-
terior cardinals. At the 10-mm. stage these upper vessels connecting with the posterior cardinals are
still both present but dwindling in importance, and the two subcardinals have begun to join across the
mid-line by a series of anastomoses. .
. This..., establishes what amounts to a single vessel which can be
 INTERNAL ANATOMY 87

Subcardinal

R L R L
Fm. 38. Development of the postcaval vein in the pig. Left, subcardinals added
to drainage of mesonephroi in 1()..mm. pig, in addition to original postcardinals;
Right, fusion of subcardinals to form postcaval. Vessels added .............. , vessels
dropped ......•

traced forward into the heart by way of a new addition to its an-
terior end. The whole channel is the developing postcaval vein.
It s~uld_Q~_noted that other vessels also contribute to the develop-
ment of the postcaval but at this stage they cannot be clearly
demonstrated.
In order to work out the postcaval vein and its relationships,
it is convenient to start with a section about midway back through
... the body, such as Fig. 36, and work forward and back in the series.
5. M esonephric Veins. Small vessels arising from the mesonephros which communicate with the
postcaval vein.
6. I ntersegmental Veins. Small vessels draining the dorsolateral body wall and flowing into the
posterior cardinals.