Physio ex reviewer:

Experiment 2

Skeletal Muscles

- Spans 2 joints and attach to skeletons thru tendons

o Tendons attach to periosteum of a bone
- Composed of hundreds to thousands of muscle fibers
o Muscle fibers produce muscle tension also known as muscle force
- When isolated and placed on a force transducer
o Possible to generate muscle contractions
Controlled electrical stimulation
o An in vitro experiments producing in vivo function

Activity 1: Muscle Twitch and Latent Period

Motor Unit

- Consists of:
o Motor neuron and Intersect at Neuromuscular
o Muscle fibers it innervates Junction (NMJ)
- Neuromuscular Junction
o Location where axon terminal of neuron meets specialized region of muscle fiber plasma membrane
Motor End Plate (MEP) specialized region
- End Plate Potential
o Events in NMJ leads to this
o Triggers a series of events that results in contraction
o Entire process called Excitation Contraction Coupling
- Action Potential
o Motor neuron triggers release of Acetylcholine (Ach) from its terminal
Ach diffused onto sarcolemma (muscle fiber plasma membrane) and binds to receptors in MEP
o Use of electrical pulses instead of Ach in expt.
- Muscle Twitch
o Single electrical stimulus will result in this
o The mechanical response to a single action potential
o Has 3 phases
Latent Period
Time that elapses between generation of Action Potential (AP) and start of contraction
No force is generated
Release of Calcium from Sarcoplasm reticulum occurs in Contraction
o Chemical changes occur intracellularly
 Contraction Phase
Starts at end of latent period and ends in peak of muscle tension
Myofilaments utilize cross-bridge cycle
Muscle develops tension
Relaxation Phase
From peak tension till end of muscle contraction
Contraction ends and muscle returns to normal resting state

Equipments used:

1. Skeletal Muscle of Frog

2. Electrical Stimulator
a. Delivers voltage via electrodes on muscles
3. Mounting stand
a. Includes force transducer
b. Measures amount of force/tension in muscle
4. Oscilloscope
a. Displays stimulated muscle twitch and amount of active, passive and total force by muscle

Things to note while doing Physio Ex Expt 2 activity 1

Pre lab Quiz

1. Skeletal muscles are connected to bones by Tendons

2. Skeletal muscles are composed of thousands of cells called Fibers
3. Motor Unit refers to 1 motor Neuron and all the Skeletal muscle fibers it innervates
4. Motor Neuron and Muscle intersect at the Neuromuscular Junction
5. A twitch is 1 contractile response to a single action potential

Predict Question:

Will changes to stimulus voltage alter the duration of the latent period?

What occurs during the latent period of these isometric contractions?

- All the steps of excitation contraction coupling occur

Activity 2: The Effect of Stimulus Voltage on Skeletal Muscle Contraction

Skeletal Muscle

- Produces tension (aka muscle force) when nervous/electrical stimulation is applied

- Number of active motor units at a given moment
o Reflected by force generated by whole muscle
- Strong Muscle Contraction = Many Motor Units activated
- Weak muscle Contraction = few motor units activated (but each unit still reaches its maximal tension)
Motor Unit Recruitment

- Increasing number of active motor units we can produce a steady increase in muscle force this is called Motor Unit
Recruitment
- As in literal na nagrerecruit ka ng motor unit para dumami chz

Muscle Twitch

- A single stimulated contraction

- Regardless of number of motor units activated, still considered 1 stimulated contraction

In this activity you will:

- Isometric/fixed - length contraction of an isolated skeletal muscle

- Involves indirect stimulation by electrode

Threshold Voltage

- Smallest stimulus required to induce AP

- Stimulus Voltage
o As stimulus voltage to muscle increases beyond threshold voltage, amount of force produced by whole muscle
increases
- Maximal tension
o Occurs when all muscle fibers have been activated by a strong stimulus (the maximal voltage)

Pre-lab Quiz:

1. Skeletal muscle fibers are innervated/stimulated by motor neurons

2. A single action potential propagating down a motor axon results in a single action potential and a single contractile event in
the muscle fibers it innervates
3. In resting skeletal muscle, Calcium is stored in the sarcoplasmic reticulum
4. During latent period for isometric contraction the cellular events involved in excitation contraction coupling occur

Stop And Think Question:

What do you see in the active force display when the stimulus voltage is set to 0.0, and why does this observation make sense?

- 0.00 g there was no activation of skeletal muscle fibers by the stimulus

Maximal voltage

- Amount of stimulus required to successfully recruit all muscle fibers into developing active force

Why was maximal voltage observed in the expt?

- At maximal voltage, all muscle fibers in muscle are depolarized all develop active force (that is they were all successfully
recruited)

Activity 3: Effect of Stimulus Frequency on Skeletal Muscle Contraction

Treppe

- Progressive increase in force generated when a muscle is stimulated in succession

- Results in muscle twitches following one another closely
o With each successive twitch peaking slightly higher than the one before
- Also known as staircase effect (because of step-like increase)
o First few twitches, successive twitches produce more force than previous twitch
As long as muscle is able to fully relax between stimuli

Skeletal muscle stimulated repeatedly

- Twitches may overlap producing a strong muscle contraction

- Phenomenon known as Wave Summation
o Wave summation occurs when muscle fibers that are developing tensions are stimulated before fibers relax
o Achieved by increasing stimulus frequency or rate of stimulus delivery to muscle
o Occurs because muscle fibers are partially contracted when subsequent stimuli are delivered

Pre-lab quiz

1. During a single twitch of a skeletal muscle, maximal force is never achieved

2. Wave summation increases the force produced in the muscle, another way to increase the force produced by a muscle is to
increase the number of activated motor units

Was there any change in the force generated by the muscle during the second stimulated twitch?

- Yes, the second twitch generated more muscle force

- This demonstrates treppe
- Despite the treppe there is a limit to the increase

Activity 4: Tetanus in Isolated Skeletal Muscle

Increasing Stimulus Frequency

- Results in increase in force produced by whole muscle

Plateau

- State known as unfused tetanus

- Happens when stimuli continue to be applied frequently to a muscle over a prolonged period of time
- The maximum possible muscle force from each stimulus will reach this

Complete (fused ) Tetanus

- Stimuli applied with greater frequency

- Twitches begin to fuse so that the peaks of each twitch become indistinguishable from one another

Maximal Tetanic Tension

- When stimulus reaches a value beyond which no further increases in force are generated by muscle
Pre-lab Quiz

1. Stimulus frequency refers to the rate that stimulating voltage pulses are applied to an isolated whole skeletal muscle
2. In a state of unfused tetanus, muscle tension increases and decreases
3. Maximal tetanic tension is when the stimulus frequency reaches a value beyond which no further increases in force are
generated by the muscle

*Fused tetanus develops at greater stimulus frequency

*Maximal tetanic tension develops with high stimulation frequencies

Post Lab:

1. The term tetanus refers to sustained muscle tension due to very frequent stimuli
2. Maximal Tetanic tension is when the stimulus frequency reaches a value beyond which no further increases in force are
generated by the muscle

Activity 5: Fatigue in Isolated Skeletal Muscle

Maximal Tetanic Tension

- When stimulus frequency reaches a value beyond which no further increase in force is generated by the muscle

Fatigue

- Refers to decline in skeletal muscles ability to maintain a constant level of force/tension after prolonged repetitive
stimulation
- Causes of fatigue (thought to be involved)
o Accumulation of lactic acid
o ADP and P in muscles from exercise
- Failure of a muscle fiber to produce tension because of previous contractile activity
- Decline in muscles ability to maintain a constant force of contraction after prolonged, repetitive stimulation

Rest Periods

- Alter onset of fatigue in skeletal muscle

Intracellular concentrations of ADP and P decline during rest period

Post-Lab

1. During cross bridge cycling in skeletal muscle force is creased by the Power Stroke of Myosin Heads
2. The term tetanus refers to sustained muscle tension due to repetitive stimuli
3. Decline in muscles ability to maintain a constant level of force or tension after prolonged repetitive stimulation is called
fatigue
4. During fatigue a numer of active cross bridges begin to decline although the rate of stimulus delivery remains constant
5. If an intervening rest period is imposed on skeletal muscle the development of fatigue will be delayed
Activity 6: The Skeletal Muscle Length Tension Relationship

Skeletal Muscle Contractions:

- Either:
o Isometric Contraction
When a muscle attempts to move a load that is equal to the force generated by the muscle
Isometric means same length
Accomplished experimentally by keeping both ends of the muscle in a fixed position while electrically
stimulating the muscle
o Isotonic Contraction

Resting Length

- Length of muscle before stimulation

- Factor in determining the amount of force that a muscle can develop when stimulated

Passive force

- Generated by stretching the muscle

- Results from elastic recoil of tissue itself
- Caused by protein titin
o Titin acts as molecular bungee cord

Active Force

- Generated when myosin thick filaments bind to actin thin filaments

o Engages the cross bridge cycle and ATP hydrolysis

Skeletal Muscle

- 2 force properties
o Passive force when it is stretched
o Active force when stimulated
- Total Force
o Sum of passive and active forces

Isometric Length Tension Relationship

- Used to understand optimum resting length in max force production

Pre-lab

1. During an isometric contraction the skeletal muscle is generating force but remains at a fixed length
2. Active force is determined by the amount of myosin bound to actin
 3. The Isometric length-tension curve indicates 3 forces the:
a. Total force
b. Active force
c. Passive force
4. Passive force in skeletal muscle is largely caused by the protein titin

Post Lab

1. The muscle is contracting isometrically when a skeletal muscle is stimulated and generates force but remains at a fixed
length
2. Titin is responsible for passive force
3. Active force stimulated through a range of muscle lengths will utilize ATP hydrolysis to drive the cross bridge cycle
4. Maximal active tension will be produced in a skeletal muscle fiber when the fiber is at its resting length

Activity 7: Isotonic Contractions and the Load Velocity Relationship

Isotonic Contraction

- Skeletal muscle length changes

- Load moves a measurable distance
- Isotonic Concentric Contraction
o Happens when the muscle length shorts as the load moves
o Happens when muscle generates a force greater than the load attached to the muscles end
- There is a Latent Period in this contraction
o There is a rise in muscle tension but no movement of weight
o After muscle tension exceeds the weight of the load an isotonic concentric contraction begins
o Latent period gets longer as the weight of the load gets larger
o Building muscle force exceeds load, muscle shortens, weight moves
- Relaxation Phase
o Force of contraction will decrease as the muscle twitch begins in the relaxation phase
o Load will return to its original position

Isotonic Twitch

- Not all-or-nothing
- Load is increased, muscle must generate more force to move it
- Latent period will get longer because it will take more time for the force to generate by the muscle

Muscle Shortening Velocity

- Speed of contraction
- Depends on the load that the muscle is attempting to move

Maximal Shortening Velocity

- Attained with minimal load attached to the muscle

- The heavier the load, the slower the muscle twitch
Isotonic Muscle Contraction Experiment

- One end of the muscle remains free

- Unlike isometric contraction (both ends are held at a fixed position)

Pre-Lab Quiz

1. During isotonic concentric contraction the force generated by the muscle is greater than the weight of the attached load
2. During isotonic concentric contraction the latent period increases with heavier loads
3. During latent period for isotonic concentric contraction cross bridges cycle and when muscle tension exceeds the load,
muscle shortening occurs
4. Muscle shortening velocity is constant regardless of load
5. The length of muscle changes in isotonic contraction

Post Lab Quiz

1. Fastest Muscle Shortening velocity = lightest load

2. Longest Latent Period of Muscle Contraction = Heaviest Load
3. Muscle Contraction move the greatest distance = Lightest load
4. Longest duration of muscle contraction = Lightest Load
5. Isotonic contraction of a muscle is one in which the length of the muscle changes
6. In lifting a heavy load the muscle shortening velocity is decreased in comparison with lighter loads