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Skeletal Muscle Physiology

INTRODUCTION
The physiology of skeletal muscle was not fully understood until the early 20th century. Some of the
earliest experiments on muscle physiology were performed between 1661 and 1665 by Jan
Swammerdam, who demonstrated that an isolated frog muscle could be made to contract when the
sciatic nerve was irritated with a metal object. Later, Luigi Galvani (1737–1798) demonstrated that frog
muscle responded to electrical currents (Fulton and Wilson, 1966).

The basic unit of a muscle is the muscle cell, or fiber. Whole muscles are made up of bundles of these
fibers. A single muscle fiber has a very regular structure, defined by myofibrils. Each myofibril consists of
an arrangement of the contractile proteins actin and myosin, which are able to slide past each other in
the presence of Ca2+ and ATP.

Figure 6.1. Skeletal muscle structure.

Skeletal muscle is similar to nerve tissue in that it responds to a stimulus in an all-or-none fashion. This
response is called a twitch. Depending on the intensity and frequency of stimulation, greater numbers of
fibers are activated. By increasing the number of active muscle fibers, the muscle is able to increase the
force it generates. Muscles with large cross-sectional areas are able to generate larger forces than those
with small cross-sectional areas.

The muscle fibers and their associated motor nerves are called a motor unit. Greater numbers of motor
nerves associated with a muscle give finer control over that muscle. Motor nerves release the
neurotransmitter acetylcholine from their synaptic bulbs onto the muscle. This junction between a

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 Exercise 6 – Physiology of Skeletal Muscle

nerve and a muscle is called the motor end plate. The release of acetylcholine at the motor end plate
depolarizes the muscle tissue and in turn leads to the release of intracellular calcium from the
sarcoplasmic reticulum, a variant of smooth endoplasmic reticulum. This release of intracellular calcium
sets in motion the biochemical events that allow actin and myosin to move past each other and
hydrolyze ATP for energy.

Figure 6.2. Skeletal muscle motor unit.

In this experiment, you will examine the basic principles of skeletal muscle physiology, including the all-
or-none response, the effect of stimulus intensity and frequency on contraction force and the
phenomenon of muscle fatigue. These experiments illustrate the collective understanding of muscle
physiology gained from over 400 years of research.

Figure 6.3. Some muscles of the forearm and hand.

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 Exercise 6 – Physiology of Skeletal Muscle

OBJECTIVES
At the end of this exercise, you are expected to:
1. demonstrate the effects of electrical stimuli using the nerves of the forearm;
2. record and measure the muscular twitch response to nerve stimulation, and show recruitment
in the twitch response as the stimulus strength increases;
3. measure the effects of changing the interval between paired stimulus pulses and observe a
short tetanic contraction;
4. calibrate a hand dynamometer with respect to a volunteer's maximal grip strength; and
5. measure the decline in maximal force during a sustained contraction, and examine some
properties of muscular fatigue.

MATERIALS
• PowerLab 15T [ML818], or PowerLab 4/26T [ML856]
• Stimulating Bar Electrode [MLADDF30]
• Electrode Cream [MLA1090]
• Finger Pulse Transducer [MLT1010]
• Hand Dynamometer [MLT003/D]

PROCEDURE
A. EQUIPMENT SETUP

Caution! Some exercises involve application of electrical shocks to muscle through electrodes
placed on the skin. People who have cardiac pacemakers or who suffer from neurological or cardiac
disorders should not volunteer for such exercises. If the volunteer feels major discomfort during the
exercises, discontinue the exercise immediately and consult your instructor.

1. Make sure the PowerLab is connected and turned on.

2. Connect the Finger Pulse Transducer to Input 1 on the PowerLab.

3. Place the Finger Pulse Transducer diaphragm-side up on the top of the lab bench, and tape the
transducer in place along the Velcro strap.

4. Connect the Stimulating Bar Electrode to the isolated stimulator output of the PowerLab.

The leads are color-coded. Plug the red lead into the red socket and the black lead into the black
socket.

5. Place a small amount of electrode cream on the two silver contacts of the stimulating bar.

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 Exercise 6 – Physiology of Skeletal Muscle

Figure 6.4. Equipment setup.

B. NERVE STIMULATION
Explore the effects of electrical stimuli using the nerves of the forearm and a Stimulating Bar
Electrode.

1. Check that the stimulator switch is off.

2. Place the Stimulating Bar Electrode over the volunteer's ulnar nerve at the wrist. The
Stimulating Bar Electrode should be held in place along the axis of the arm with the red dot
closest to the elbow.

Figure 6.5. Placement of the stimulating bar electrode.

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 Exercise 6 – Physiology of Skeletal Muscle

3. In the Stimulator panel set current to 5 mA.

4. Click Start then set the stimulator switch to ON. The stimulator status light should now flash
green, indicating that the chosen stimulus current is being passed through the subject's skin. If
the light flashes yellow, current is not flowing properly.
5. Note the twitch contractions affecting the thumb and fingers. Examine the effect of small
adjustments in the position of the electrodes, and locate the position giving the largest twitches.
If no twitch occurs, increase the stimulus current.

6. Explore the motor and sensory results of stimulating at other places in the forearm. Each time
you move the electrode to another location, wipe away the residual electrode cream from the
skin to prevent short-circuiting. (Stimulation will be ineffective if the current flows along a
surface layer of electrode cream rather than through the arm.)

You will probably find that effective stimulation will only occur when the two pads of the bar
electrode are aligned along the arm's length. If the stimulus status light changes in color from green
to yellow, you will need to put more electrode cream on the pads.

7. Try stimulating the ulnar nerve at the level of the elbow. The nerve passes behind a bony
prominence (the medial epicondyle) on the humerus. At this location, the nerve is exposed to
minor mechanical injury and is known to children as the "funny bone". Stimulation at this site
gives large and obvious motor effects.

8. Click Stop and turn the stimulator switch off.

C. TWITCH RESPONSE AND RECRUITMENT

Record and measure the muscular twitch response to nerve stimulation, and investigate recruitment as
the stimulus strength increases.

FINDING THE THRESHOLD

1. Have the volunteer place his or her hand as shown here, with the fingers under the edge of the
table and the edge of the thumb resting lightly on the pulse transducer. (If the table edge is too
thick for the subject's hand, a plank or shelf may have to be used.)

2. Wipe the electrode cream from the subject's wrist.

3. Apply a small amount of electrode cream to the pads of the stimulating bar electrode.

4. Make the subject hold the electrode with the free hand firmly in place at the site for stimulation
of the ulnar nerve at the wrist. Ensure that the edge of the subject's thumb is resting lightly on
the transducer.

5. Ensure that the stimulus current is set to 1 mA.

6. Set the stimulator switch to ON.

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 Exercise 6 – Physiology of Skeletal Muscle

7. Click Start.

LabTutor will stimulate, record for a fixed duration of 0.5 seconds and then stop automatically.

8. Increase the stimulus current to 2.0 mA.

9. Click Start.

10. Continue to increase the stimulus current in 1 mA steps, clicking Start each time.

For most subjects, the threshold stimulus at which a response is first observed is in the range 3-
8 mA.

11. When you first see a response, add a comment to the recording, noting the subject's name and
the stimulus current used.

RECRUITMENT

12. Reduce the amplitude by 1 mA.

13. Click Start.

14. Increase the amplitude in 0.5 mA steps, clicking Start each time and adding a comment, noting
the current used.

15. Continue this until the response no longer increases.

For most subjects, this maximal stimulus is in the range 6-15 mA.

ANALYSIS

1. Locate the beginning of the recruitment recordings. Type into the first column of the table the
current delivered to produce each response.

2. Move the cursor over the waveform and click on the peak of each response to transfer its value
to the Value panel.
3. Drag the value to the appropriate cell in the table.

As you enter the data, the Graph panel will graphically display the relationship between stimulus current
and response size.

Note the stimulus current at which the response no longer increases. This is called the 'maximal
stimulus'

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 Exercise 6 – Physiology of Skeletal Muscle

D. SUMMATION

Now stimulate with paired pulses, and investigate how varying the interval between the pulses affects
the response.

Use the same setup as in Exercise 2.

1. Turn the Stimulator switch ON.

2. In the Stimulator panel, set the current to 5 mA greater than the maximal stimulus you
determined in Part C.

Be sure to use the proper value for the present subject!

3. Check that the stimulus interval is set to 1000 ms.

4. Click Start.

LabTutor will automatically deliver the stimulus and stop recording after a fixed duration of 3
seconds.

5. In the Stimulator panel decrease the stimulus interval to 500 ms, and click Start.

6. Repeat this for intervals 200 ms, 150 ms, 100 ms and 50 ms, noting the values in comments as
you did above.

When you have finished, turn the Stimulator switch OFF, ready for the next exercise.

ANALYSIS

1. Place the Marker on the baseline and the Waveform Cursor on the Peak to determine the peak
amplitude of the first response.

2. Click on the peak to enter the force value into the Value panel.

3. Drag the value into the appropriate cell in the table.

4. Repeat steps 1-3 above for each response, at each stimulus interval.

E. TETANUS

Examine the effect of rapid stimulation and observe a short tetanic contraction.

1. Ensure that the volunteer's hand and stimulus electrodes are placed as before, and turn the
stimulator switch ON.

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 Exercise 6 – Physiology of Skeletal Muscle

2. Check that the stimulus interval is set to 50 ms. Set the number of pulses to 1.

3. Click Start.

LabTutor will automatically deliver the stimulus and stop recording after a fixed duration of 3
seconds.

4. Change the number of pulses to 2 and stimulate again.

5. Change the number of pulses to 3 and stimulate again.

6. If there was not too much discomfort, increase the number of pulses to 4 or 5 depending on the
comfort level of the subject.

7. Turn the stimulator switch OFF and disconnect the stimulating bar electrode and the finger
pulse transducer from the PowerLab.

ANALYSIS

1. Place the Marker on the baseline and the Waveform Cursor on the Peak to determine the peak
amplitude.

2. Click on the peak to enter the force value into the Value panel.

3. Drag the value into the appropriate cell in the table.

F. GRIP FORCE CALIBRATION

In the next exercise we investigate the decline in maximal force during a sustained contraction. Be sure
the same volunteer performs both the calibration and the exercise to achieve accurate results.

Make sure that you have removed the finger pulse transducer and the electrodes from the PowerLab.

1. Connect the plug of the grip force transducer to the Input 1.

2. The volunteer should loosely grip the hand dynamometer in the fist, as shown here.

3. Click Start.

4. The volunteer should squeeze the dynamometer as hard as possible for a second or two, and
then relax.

5. Click Stop

Now, to calibrate for the strength of the volunteer:

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 Exercise 6 – Physiology of Skeletal Muscle

6. Click the trace at a time when the force is effectively zero, then click the Point 1 button in the
Calibration panel. This will set the 0% grip force.

7. Click the trace at peak force, and click the Point 2 button in the Calibration panel. This
represents 100% of the subject's grip force.

8. Click Apply.

G. FATIGUE

Measure the decline in maximal force during a sustained contraction, and examine some properties of
muscular fatigue

The grip force transducer should already be calibrated for the volunteer, as described in the
previous page.

1. Allow the volunteer to view the computer screen.

2. Click Start.

3. Ask the volunteer to maintain 25% maximal grip strength while watching the recorded trace.

4. After 20 seconds, tell the volunteer to relax.

5. Click Stop.

6. Wait for 30 seconds to allow recovery of muscle function.

7. Repeat steps 2-6 for contractions of 50%, 75% and 100% of maximal grip strength.

8. Allow the volunteer to rest for two minutes.

9. Turn the volunteer away so that he or she cannot see the computer screen.

10. Click Start.

11. Ask the volunteer to produce a sustained maximal contraction.

12. After 8 to 10 seconds, or when the force has obviously declined, instruct them to try harder.

13. After a further 8 to 10 seconds, repeat the encouragement.

14. A few seconds later, ask the volunteer to relax.

15. Click Stop.

16. Click Start.

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 Exercise 6 – Physiology of Skeletal Muscle

17. Ask the volunteer to produce a sustained maximal contraction. Every 8 to 10 seconds, allow the
volunteer to relax very briefly (half a second), and then return to maximal contraction.

18. After 30 to 40 seconds click Stop.

19. Allow the volunteer to use his or her other hand if gripping the transducer has become painful.
Turn the volunteer so that they can see the computer screen.

20. Click Start.

21. Ask the volunteer to produce a 50% contraction while watching the trace.

22. After 10 seconds, press the Enter key to enter a comment (to mark the time).

23. Have the volunteer close his or her eyes, and attempt to maintain exactly the same contraction
force for the next 30 seconds.

24. After the elapsed time, the volunteer should open their eyes, and adjust the contraction force
back to 50%.

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