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Experiment 2: Skeletal Muscle Physiology

Camille Butac, Nickey Angelo Cayetano, Kersten Calimag, Ma. Audrey Jannie Dalupang

Department of Biological Sciences

College of Science, University of Santo Tomas
España, Manila, Philippines

A muscle contraction involves a wave of action potential in a motor neuron which triggers the release of
calcium ions from the sarcoplasmic reticulum of striated muscle cells. It is the calcium ions that play a vital role in
muscle contractions triggering actin, and myosin cross-bridge formation. In the first experiment, an isolated striated
muscle is stimulated to demonstrate treppe, and wave summation. Treppe was observed when there was an increase
in the active force generated by the muscle when subsequent stimulus, with intervening rest, occurs. The total force
generated has increased from 1.83 g to 2.40 g when subsequent stimulus occurs. Wave summation was observed
when multiple stimulus, with no intervening rest, increases the total force generated. Multiple stimulus has increased
the total force from 1.83 g to 2.72, 3.20 or even as high as 5.57 g. Furthermore, an increase in the voltage stimulus
also increases the total force generated by the muscle contraction. In the second experiment, an isolated striated
muscle is stimulated to demonstrate the relationship between the length and the force generated by the muscle. With
decreasing length, the active force generated decreases. When the length of the muscle is at 75 mm, an active force
of 1.82 g occurs. Decreasing the length to about 50 mm would decrease the active force to about 0.11 g. In contrary,
increasing the length would increase the active force, and generate a passive force. An increase in muscle length to
80 mm would generate an active force of 1.75 g and a passive force of 0.02 g. In the third experiment, an isolated
striated muscle is stimulated to demonstrate the effect of load to muscle speed and contracted distance. With
increasing load, there would be a subsequent decrease in muscle contraction velocity, twitch duration, and
contracted distance. A 0.5g load would have a muscle velocity of 0.100 mm/msec, a twitch duration of 78 msec, and
a contracted distance of 4 mm. A 2.0 g load, on the other hand, would have a muscle velocity, twitch duration, and
contracted distance of 0.

Skeletal muscles are the reason behind our ability to move our bodies. These are striated
muscles that characteristically span two joints and adhere by means of tendons, which append to
the periosteum of the bone. Based on its innervation, it is classified as voluntary muscle which
composes of individual cells termed muscle fibers that produce muscle tension also known as
muscle force. These muscles are able to contract by the active sliding of the thick and thin
filaments past each other as interpreted by the sliding filament mechanism. Moreover, a motor
unit includes a motor neuron, its axon fibers and all the muscle fibers it stimulate. An all-or-none
response is exhibited by a single muscle fiber wherein it contracts when stimulated by a
threshold stimulus.
An isolated skeletal muscle of an experimental animal can produce muscle contractions
when mounted on a force transducer with controlled electrical stimulation. Correspondingly, the
isolated muscle imitate the contractions of those which are active in the body. A single stimulus
results in a muscle twitch; this is the mechanical response brought about by a single action
potential. The three phases of a muscle twitch include: latent period. contraction period,
relaxation period.
In the effect of stimulus frequency on contraction, treppe is the continuous escalation in
force generated when there is a successive stimulation. Treppe is also known as the staircase
effect because of its step-like increase in force. On the other hand, wave summation arises when
the muscle fibers developing tension are repeatedly stimulated even before the fibers have
relaxed. This is achieved by increasing the rate of stimulus delivery to the muscle, or
stimulus frequency. Muscle contractions are either classified as isometric or isotonic.
Isometric contractions are experimentally carried out by keeping the muscles’ both ends
in a fixed position while it is electrically stimulated. In addition, a muscle contracts isometrically
when the force generated by the muscle is equal to the load it attempts to move. Passive force
results from the elastic recoil of the of the stretched muscle tissue itself. The generation of active
force is related to the cross bridge cycle and ATP hydrolysis when myosin binds to actin.
Contrastingly, isotonic contraction occurs when the force generated is greater than the load
adhered to the muscles’ end. In this type of contraction, the muscle length shortens and therefore,
the load involves movement.


Activity 3: The Effect of Stimulus Frequency on Skeletal Muscle Contraction

Using an isolated skeletal muscle and a stimulator, the stimulus was set to 8.5 V and the muscle
was first stimulated and traced using an oscilloscope, this is the first trial. After every trial, the
results were recorded. In the second trial, the muscle was stimulated again allowing the trace to
rise and fall before stimulating a second time. In the third trial,the muscle was stimulated with
two successive stimulus. In the fourth trial, the muscle was stimulated with four successive
stimulus. In the fifth trial, the stimulus was increase to 10 V with multiple stimulus. In the last
trial, the stimulus is reverted to 8.5 V and there is no limit to the stimulus applied.

Activity 6: The Skeletal Muscle Length-Tension Relationship

The resting muscle was set to 75 mm while the voltage was set to 8.5 volts. In order to deliver an
electrical stimulus to the muscle, the stimulate button was employed. Afterwards, three data
points were presented representing the active, passive, and total force generated during the
twitch. On the left oscilloscope display, a tracing of a single muscle twitch was seen. The muscle
was shortened by 5mm to determine the effect of muscle length on the different forces. The
stimulate button was again applied to note the values of the forces in comparison to the original
75 mm. These steps were repeated until a muscle length of 50 mm was reached. Eventually,
when the muscle was lengthened to 80 mm, same procedures were applied in delivering
electrical stimulus. Subsequently, the muscle was gradually lengthened by 10mm and was
stimulated until a length of 100mm was reached. The recordings were compared to the original
75mm muscle length. At the end of the experiment, the plot data showed the summary of the
recorded data.

Activity 7: ​Isotonic Contractions and the Load-Velocity Relationship

A 0.5-g weight was dragged from the cabinet and attached to the free end of the muscle. Then an
electrical stimulus to the muscle was delivered. The muscle action was then observed. As the
muscle shortened in length it lifted the weight off the platform. After, the muscle elongated as it
is relaxed and lowered the weight back down to the platform. The muscle was stimulated again
this time, both the muscle and oscilloscope screen was observed at the same time. The data was
then recorded.The current 0.5 g weight was removed and was replaced by 1.0-g weight. The
same procedure was performed with the other weights (1.5-g and 2.0-g). Lastly, the data
recorded was plotted.

Activity 3: The Effect of Stimulus Frequency on Skeletal Muscle Contraction

Table 1. The Effect of Voltage and the Frequency of Stimulus on the Active, Passive, and Total Force Generated by the Muscle

Graph 1.1. Single Stimulus Graph 1.2. Two Subsequent Stimulus with Rest
Graph 1.3. Two Subsequent Stimulus without Rest Graph 1.4. Four Subsequent Stimulus without Rest

Graph 1.5. Increasing Voltage Graph 1.6: Multiple Stimulus without Rest

There are three types of graded muscle responses: motor recruitment, wave summation,
and treppe. The latter two are the ones focused in this experiment. Wave summation happens
when muscle fibers that are developing tension are stimulated again before the fibers have
relaxed. It is predominantly attained by increasing the stimulus frequency, or the rate of stimulus
transport to the muscle. Treppe is the gradual increase in force generated when a muscle is
stimulated in progression, such that muscle twitches follow one another closely, with each
succeeding twitch peaking marginally higher than the one beforehand. If stimulus frequency
increases, the force generated with each successive stimulus similarly increases. This is both true
for wave summation and treppe. The solitary distinction is that, treppe permits the muscle to
relax before another stimulus is generated while in wave summation, stimulus is applied before
the muscle can even relax. Also, both have a maximum or threshold to the force generated.
Hence, the amount of active force created can be increased not only by increasing the stimulus
intensity but also the stimulus frequency.

Activity 6: ​The Skeletal Muscle Length-Tension Relationship

Table 2. The Effect of Muscle Length to the Active, Passive, and Total Force Generated by the Muscle

Graph 2.1: Muscle Length at 75 mm with Plot Data (right).

Graph 2.2. Muscle Length at 70 mm with Plot Data (right). Subsequent Graph and Plot Data Shows Preceding Data
Graph 2.3: Muscle Length at 65 mm with Plot Data (right)

Graph 2.4. Muscle Length at 60 mm with Plot Data (right)

Graph 2.5. Muscle Length at 55 mm with Plot Data (right)

Graph 2.6. Muscle Length at 50 mm with Plot Data (right)

Graph 2.7. Muscle Length at 80 mm with Plot Data (right)

Graph 2.8. Muscle Length at 90 mm with Plot Data (right)

Graph 2.9. Muscle Length at 100 mm with Plot Data (right)

Graph 2.10. Summary of Active Force Data

Based on the muscle length vs tension curve presented, there is a definite length range
where a skeletal muscle produces the highest tension which is the optimum latent length. This
correlation refers to the length-tension relationship. In the data table, 50 is the shortest length and
was able to produce the lowest tension whereas the longest length, 90, generated a higher tension
but lower compared to a length that is somewhere in the middle of the range. At a length of 50,
there would be little or no tension will be generated for the lack of space, as provided by the z
lines, for the myosin heads to move. At the optimal length, 75, it is such because the myosin
heads in contact with actin filaments are many, or close to all, generating a substantial amount of
tension. The passive force in lengths higher than 75 is produced by the protein titin due to its
spring-like nature.
Activity 7: ​Isotonic Contractions and the Load-Velocity Relationship

Table 3. The Effect of Load to the Speed of Muscle Contraction and the Distance of the Muscle Contracted

Graph 3.1. Muscle Contraction with 0.5 g Load

Graph 3.2 Muscle Contraction with 1.0 g Load

Graph 3.3. Muscle Contraction with 1.5 g Load

Graph 3.4. Muscle Contraction with 2.0 g Load

Graph 3.5. Muscle Load-Velocity Relationship

Isotonic muscle contraction creates force when muscle length is shifting. There are two
kinds of isotonic muscle contraction explicitly concentric muscle contraction and eccentric
muscle contraction. Concentric muscle contraction triggers the muscle to shorten thus producing
force. Instead, an eccentric muscle contraction triggers the muscle to extend due to greater
opposite force. In this experiment, as the load on the muscle increases, the latent period will
correspondingly increase, however the distance will decrease, as well as the contraction duration.
These are all distinct from the data in the table above. All of the weights used are isotonic
concentric contractions because the muscle varied in length contrasting to the one with a 2.0
-gram weight where the length of the muscle persisted the identical thus it can be classified as an
isometric contraction.
Muscle contraction results as a wave of action potential passes through a motor neuron.
There are a number of factors which affect muscle contraction. Three of which are the frequency
of stimulation, the length of muscle, and the load carried by the muscle. An increase in the
frequency of stimulation would lead to an increase in the total force generated by the muscle. An
increase in the length of the muscle would lead to an increase in the active force and a
subsequent increase in the passive force. Finally, an increase in load would lead to a decrease in
the speed of contraction, twitch duration, and distance contracted by the muscle. In other words,
there is a positive relationship between the frequency of stimulation and the length of the muscle
to the total force generated by that muscle, while there is a negative relationship between the
load carried by the muscle to the velocity of contraction.