Neurobiological Basis of Sleep

Julie Ann Kristy L. Torres, MD, FPCP, FPNA

Brokenshire College – School of Medicine
References
 Why do we sleep and wake up?
• Humans evolved as equatorial
animals and our behavior is
dominated by a circadian rhythm
of daylight activity and nocturnal
sleep
 Biological Rhythms
• Defined as the inherent timing mechanisms that control or
initiate various biological processes
– Linked to cycles of days and seasons produced by the Earth’s rotation
on its axis
 Biologic Clock
• Synchronize the behavior of organisms to the temporal passage of a real
day
• Allows an organism to anticipate events in advance and prepare for
them both physiologically and cognitively
• Regulates feeding times, sleeping times and metabolic activity so that
they are appropriate to day-night cycles
• Regulate gene expression in every cell for homeostasis
 Biologic Clock
• 24 hours
– Time required to complete a cycle of activity
 Suprachiasmatic Nucleus
• Main biological clock
• Located in the hypothalamus
– Above the optic chiasm
• Curt Richter (1965)
– First to locate the SCN
– Circadian rhythm in rats lost when
SCN was ablated by electrical
current
 Suprachiasmatic Nucleus
• Neurons in this area have an
intrinsically rhythmic pattern of
activity that is genetically
determined
 Suprachiasmatic Nucleus
• GABA
– main neurotransmitter of the SCN neurons
– Inhibitory synapses
– Allows SCN neurons to act in synchrony
 Suprachiasmatic Nucleus
• 2 parts:
1. Core – non rhythmic,
entrains the shell
neurons
2. Shell - rhythmic
 Suprachiasmatic Nucleus
• Zeitgebers
– Time-givers or cues that entrain the SCN neurons to the solar day
– Light as the main zeitgeber
– Others:
• feeding, mobility, arousal, changes in lighting
 Suprachiasmatic Nucleus
• Experiment in rodents:
– When a radioactive tracer is injected into rodents, more tracer is
found in the SCN after injections given in the light period than after
injections given in the dark period of the light–dark cycle
– Therefore, suprachiasmatic nucleus neurons are more active during
the light period.
Suprachiasmatic Nucleus

LIGHT ON LIGHT OFF

 Suprachiasmatic Nucleus
• Light entrains the SCN and in turn drives a number of slave
oscillators, which are responsible for rhythmic occurrence of one
activity
 Suprachiasmatic Nucleus
• The SCN clock entrains slave oscillators through:
1. SCN neurons send axonal connections to nuclei close by in the
hypothalamus and thalamus.
2. The SCN connects with pituitary endocrine neurons to control the
release of a wide range of hormones.
3. The SCN also sends indirect messages to autonomic neurons in the
spinal cord to inhibit the pineal gland from producing the hormone
melatonin, which influences daily and seasonal biorhythms.
 Suprachiasmatic Nucleus
• The SCN controls the release of these hormones:
1. Melatonin
• from the pineal gland
• circulates during the dark phase of the circadian cycle
• rest activities
2. Glucocorticoids
• from the adrenal gland
• circulate during the light phase of the circadian cycle
• arousal activities
 Melatonin
• Hormone that induces
sleepiness
• Postganglionic retinal nerve
fibers mediate its production and
secretion through the
retinohypothalamic tract to the
SCN then to the superior cervical
ganglion and finally to the pineal
gland
 Melatonin
• Light inhibits its release
• Darkness facilitates its release
• Tryptophan – source of
melatonin
• The daily rhythm of melatonin
secretion is also controlled by an
endogenous, free-running
pacemaker located in the
suprachiasmatic nucleus.
 Melatonin Production
activation of a 1-and b 1-adrenergic
receptors in the pineal gland

raises cyclic AMP and calcium

concentrations

activates arylalkylamine N-
acetyltransferase, initiating the
synthesis and release of melatonin
 Biologic Clock
• Other biological clocks:
– Intergeniculate leaflet of thalamus
– Pineal gland
– Raphe nucleus of brainstem
 Damage to Suprachiasmatic Nucleus
• Animals still eat, drink, exercise, and sleep, but they do these
activities at haphazard times.
– If all the pathways into and out of the SCN are cut, SCN neurons
maintain their rhythmic electrical activity.
 Suprachiasmatic Nucleus
• The SCN receives information about light through the
retinohypothalamic tract
– This pathway begins with specialized retinal ganglion cells (RGCs)
 Retinohypothalamic Pathway
• Retinal ganglion cells
– Distributed throughout the retina
– Contains melanopsin
• photosensitive pigment
– Receive light-related signals from the rods [dim] and cones [light] and
send these signals to visual centers in the brain
– Can also be activated directly by certain wavelengths of blue light in
the absence of rods and cones
 Retinohypothalamic Pathway
• Retinal ganglion cells
– Glutamate
• main neurotransmitter
– Other neurotransmitters:
• substance P
• pituitary adenylate cyclase-activating polypeptide (PACAP)
Retinohypothalamic Pathway
Retinohypothalamic Pathway
 Molecular Basis of the Biological Clock
Step 1: Transcription

In the cell nucleus:

three Period genes (Per1,*
Per2, Per3)
two Cryptochrome genes
(Cry1, Cry2)

Transcribed into
Per1, Per2, and Per3 mRNA transcription-translation-
Cry1 and Cry2 mRNA inhibition feedback loop
 Molecular Basis of the Biological Clock
Step 2: Translation

In the endoplasmic reticulum,

ribosomes translate these
mRNAs into the proteins PE R1,
PE R2, PE R3 and CRY1, CRY2.

transcription-translation-
inhibition feedback loop
 Molecular Basis of the Biological Clock
Step 2: Translation

In the intracellular fluid, the

proteins then form various
dimers, or two protein
combinations, such as PERCRY.

transcription-translation-
inhibition feedback loop
 Molecular Basis of the Biological Clock
Step 3: Inhibition

PERCRY dimers enter the cell

nucleus, where they bind to
and inhibit the CB dimer
(formed by the CLOCK and
BMAL proteins).

transcription-translation-
inhibition feedback loop
 Molecular Basis of the Biological Clock
Step 3: Inhibition

The CB dimer turns ON the

Enhancer box (Ebox), a part of the
DNA that activates transcription of
the Period and Cryptochrome
genes.

So when the CB dimer is inhibited,

the Per and Cry genes are no
longer expressed. transcription-translation-
inhibition feedback loop
 Molecular Basis of the Biological Clock
Step 4: Decay

After they play their inhibitory role,

the PERCRY proteins decay.

Then the CB dimer resumes its

activity, the Per and Cry genes
resume expression, and the 24-
hour cycle begins anew.

transcription-translation-
inhibition feedback loop
 Molecular Basis of the Biological Clock
• This sequence of gene turn-on followed by gene turn-off
occurs in an inexorable, daily loop.
 Molecular Basis of the Biological Clock
• Mutations in any circadian gene can lead to circadian
alterations, including absence of a biorhythm or an altered
biorhythm.
– For example, alleles of Period 1 and Period 2 genes determine
chronotype.
• whether an individual will be “early to bed and early to rise” or “late to bed
and late to rise”
PHYSIOLOGY OF SLEEP
 Function of Sleep
• Not a passive process
• Biological adaptation
– Energy conserving strategy to cope with times when food is scarce
– Increases alertness in waking state
 Function of Sleep
• Neural maturation
• Facilitation of learning and memory
• Targeted erasure of synapses to “forget” unimportant
information that might clutter the synaptic network
• Cognition
• Clearance of metabolic waste products generated by neural
activity in the awake brain
• Conservation of metabolic energy
 Sleeping Behavior
• Consists of
– Resting
– Napping
– Long bouts of sleep
– Various sleep-related events including snoring, dreaming, thrashing
about, and even sleepwalking
 Cellular Basis of Sleep
• Sleep promoting activity
– Mediated by GABA through its interactions with the GABA-A receptor
• GABA released by sleep-promoting neurons in the anterior hypothalamus
– Adenosine also promotes sleep
• inhibits hypocretin/orexin neurons in the basal forebrain, lateral
hypothalamus and tuberomammillary nucleus
• Activates neurons in the preoptic/anterior hypothalamus and ventrolateral
preoptic area
 Cellular Basis of Sleep
• Wakefulness promoting activity
– Acetylcholine, dopamine, norepinephrine, serotonin, histamine and
hypocretin promote wakefulness
 Stages of Sleep
• Sleep is not a unitary state but consists of a number of stages
1. Waking state
2. Non rapid eye movement sleep – N1, N2, N3
3. Rapid eye movement sleep
• 75% of sleep is spent in NREM stage (N2)
 Stages of Sleep
• The body cycles through all stages of sleep approximately 4 to
6 times each night
– From N1 to N2 to N3 to N2 then REM
• A complete cycle takes about 90 to 110 minutes
• First REM period is short and as night progresses, longer period
of REM and decreases time in deep sleep (NREM) occur
 Rapid Eye Movement vs Non-REM Sleep
REM Sleep Non-REM Sleep
Fast wave activity Delta rhythm or slow wave
sleep

Eyes flickering back and

forth (REM)

Atonia [no muscle

tone]
 Stages of Sleep
• Awake
– EEG:
• beta rhythm – open eye awake state
• alpha rhythm – if relaxed and drowsy, eyes closed
 Stages of Sleep
• N1 Stage
– 5% of total sleep time
– Lasts for 1 to 5 minutes
– EEG: theta rhythm – low voltage
– Lightest stage of sleep
– Begins when more than 50% of alpha rhythm are replaced by low
amplitude mixed frequency activity
– Muscle tone is present, breathing is regular
 Stages of Sleep
• N2 Stage
– 45% of total sleep time
– Lasts for 25 minutes in the 1st cycle and lengthens with each
successive cycle
– EEG: sleep spindles and K complexes
– Deeper stage of sleep
– Heart rate and body temperature drop
– Bruxism may occur
 Stages of Sleep
• N3 Stage
– 25% of total sleep time
– Slow wave sleep (SWS)
– Deepest stage of sleep
– EEG: delta rhythm – lowest frequency, highest amplitude
– Difficult to awaken from; if awoken, may have sleep inertia (mental
fogginess)
– As people age, they spend less time in this stage and more time in N2
– Body repair, builds bone and muscle, strengthen immune system
– Sleepwalking, night terrors and bedwetting may occur
 Stages of Sleep
• REM Stage
– 25% of total sleep time
– Lasts for 90 minutes after sleep onset
– 1st REM cycle lasts 10 minutes and final cycle lasts for 1 hour
– EEG: beta rhythm – similar to wakefulness
– Associated with dreaming
– Atonic skeletal muscles except for eye and diaphragm movement
– Breathing is erratic and irregular
– Dreams, nightmares, penile or clitoral tumescence may occur
 Stages of Sleep
• REM Stage
– Important characteristics:
• People tend to awaken spontaneously in the morning during an episode of
REM sleep
• Loss of motor tone, increased brain O2 use, increased and variable pulse and
blood pressure
• Increased levels of ACh
• The brain is highly active throughout REM sleep, increasing brain metabolism
by up to 20%
 Evolution of Sleep through Aging
• Newborns and Infants (birth to 1 year)
– Sleep timing in newborns is distributed evenly across day and night
for the first few weeks of life, with irregular sleeping and waking
patterns.
– Newborns sleep approximately 16 to 18 hours per day
discontinuously, with the longest continuous sleep episode typically
lasting 2.5 to 4 hours.
 Evolution of Sleep through Aging
• Newborns and Infants (birth to 1 year)
– Newborns have 3 different types of sleep: quiet sleep (similar to
NREM), active sleep (similar to REM), and indeterminate sleep.
– In contrast to children and adults, newborn sleep onset occurs
through REM, not NREM, with each sleep episode consisting of
only 1 or 2 cycles.
 Evolution of Sleep through Aging
• Newborns and Infants (birth to 1 year)
– Circadian rhythms develop around 2 to 3 months of age
– At 2 months of age, the progression of nocturnal sleeping begins.
– At 3 months of age, the cycling of melatonin and cortisol in a
circadian rhythm occurs, and sleep onset begins with NREM.
• REM sleep decreases and shifts to the later part of the sleep cycle.
• The total NREM and REM sleep cycle is typically 50 minutes instead of the
adult 90-minute cycle.
 Evolution of Sleep through Aging
• Newborns and Infants (birth to 1 year)
– At 6 months of age, the longest continuous sleep episode lengthens
to 6 hours.
– At 12 months, infants typically sleep 14 to 15 hours daily, with most
sleep occurring in the evening and only 1 to 2 naps needed during
the day.
 Evolution of Sleep through Aging
• Toddlers (1 to 3) and Children (3 to 9)
– Around 2 to 5 years of age, the total sleep time needed each day
decreases by 2 hours, from 13 to 11 hours.
– By 6 years of age, children manifest circadian sleep phase
preferences and tend toward being night owls or early risers.
 Evolution of Sleep through Aging
• Adolescents (10 to 18)
– The total sleep time required for adolescents is 9 to 10 hours each
night.
– Due to various pubertal and hormonal changes accompanying
puberty's onset, slow-wave-sleep and sleep latency time declines,
and time in stage N2 increases.
– Around mid-puberty, daytime sleepiness occurs more frequently than
at earlier puberty stages.
 Evolution of Sleep through Aging
• Adults (19 years and beyond)
– Adults tend to demonstrate earlier sleep time, wake time, and
reduced sleep consolidation.
– Adults aged 65 and older awaken approximately 1.5 hours earlier
and sleep an hour earlier than adults aged 20 to 30.
Most people sleep less as they grow older
 Evolution of Sleep through Aging
• Gender differences
– Men tend to spend a greater amount of time in stage N1 sleep and
experience more nighttime awakenings, so there is a greater
propensity for daytime sleepiness.
– Women maintain slow-wave sleep longer than men and tend to
complain more often of difficulty falling asleep.
• Additionally, daytime sleepiness increases during pregnancy and the first few
months postpartum.
 Neural Basis of Sleep
• Reticular activating system
– Proposed by Moruzzi and
Magoun
– Sensory pathways entering the
brainstem have collateral
axons that synapse with
neurons in the RAS.
 Neural Basis of Sleep
• Reticular activating system
– They proposed that sensory stimulation is conveyed to RAS neurons
by these collaterals then RAS neurons produce the desynchronized
EEG via axons that project to the cortex.
 Neural Basis of Sleep
• Stimulation of RAS
– Waking EEG

• Damage to RAS
– Slow-wave, sleep-like EEG
 Neural Basis of Sleep
• Basal forebrain
– contains large cholinergic cells
which secrete acetylcholine (ACh)
from their terminals onto
neocortical neurons to stimulate a
waking EEG (beta rhythm).
– Waking associated with being still
yet alert
 Neural Basis of Sleep
• Median raphe
– Located in the midbrain
– Contains serotonin (5-HT) neurons
whose axons also project diffusely
to the neocortex, where they also
stimulate neocortical cells to
produce a beta rhythm, recorded as
a waking EEG.
– Waking associated with movement
 Neural Basis of REM Sleep
• Peribrachial area
– Responsible for REM sleep
– Dorsal part of brainstem anterior to
cerebellum
– Extends to the medial pontine
reticular formation
– Damage to this area reduces or
abolishes REM sleep
 Neural Basis of REM Sleep

SLD/subcoerulear nucleus
Clinical Correlates
 SLEEP APNEA
• Individuals with sleep apnea experience airway collapse in deeper sleep
states, causing them to experience reduced time in stage N3 and REM
sleep.
• This leads to excessive daytime drowsiness as proper, efficient sleep is
not obtained throughout the night.
• Treatment:
– CPAP or BiPaP machine during sleep
 SLEEP APNEA
• 2 types:
– Central sleep apnea occurs when
the brain fails to signal respiratory
muscles during sleep.
– Obstructive sleep apnea is a
mechanical problem in which there
is a partial or complete blockage of
the upper airway.
 REM SLEEP DISORDER
• If the temporary atonia of REM
sleep is disturbed, it may be
possible to physically act out (often
unpleasant) dreams with
vocalizations and sudden limb
movements.
• The cause of this disorder is not
entirely known but may be
associated with degenerative
neurological conditions such as
Parkinson disease or Lewy body
dementia.
 NARCOLEPSY
• Narcolepsy is a sleep cycle disorder in which individuals present with
persistent daytime sleepiness and brief episodes of muscle weakness
(cataplexy).
• Sleep regulation is disturbed, and individuals tend to skip the initial phases of
sleep and fall directly into REM sleep.
– These individuals can enter the REM phase and have dreams during short naps. This
limits their amount of sleep in the N3 deep-sleep stage and thus causes an irregular
sleep pattern.
– May experience a sudden loss of muscle strength as body muscles are atonic and
paralyzed in the REM-sleep phase.
– These lapses into REM sleep can happen anytime during the day and usually last seconds
to minutes.
 SOMNAMBULISM
• Sleepwalking
• Common occurrence in school-aged children
– These individuals tend to make purposeful movements, but they are not acting
out their dreams.
– Typically associated with common behaviors, such as dressing, eating, and
urinating.
• Occurs in the non-rapid eye movement phases, usually in N3
• Sleepwalking occurs because the sleep cycle is still in the maturing
phase, and proper sleep/wake cycles are not yet regulated.
Thank You