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2 EXERCISE Skeletal Muscle Physiology Advance Preparation/Comments 1. Prior to the lab, suggest to the students that they become familiar with the exercise before coming to class. If students have a home computer, or access to a computer on campus, they can become familiar with the general operation of the simulations before coming to class. 2. You might do a short introductory presentation with the following elements: • Describe the basics of muscle contraction at the cellular level, focusing on the sarcomere. This explanation is especially important for the isometric part of the simulation. • Students often have problems distinguishing between in vivo stimulation via the nervous system versus the electrical stimulation we apply to whole skeletal muscle in an experiment. Mention that increasing the intensity of an electrical stimulus to the surface of whole muscle is not the same as stimulation via the nervous system, but that the outcome of increased force production is similar in both methods. • Encourage students to try to apply the concepts from the simulation to human skeletal muscles as they work through the program. • If a demonstration computer screen is available, briefly show students the basic equipment parts. 3. Keep in mind that many students in an introductory science course are deficient in their graphing skills. Reviewing the principles of plotting before the class begins may prove helpful. 4. Be prepared to help the students answer the more difficult “What if . . . ” questions. Answers to Questions/Experimental Data Pre-lab Quiz in the Lab Manual 1. Tendons 2. b. A motor neuron and all of the muscle cells it innervates 3. a. Latent 4. acetylcholine 5. True 6. False 7. d. Threshold voltage 8. b. Isometric Activity 1: The Muscle Twitch and the Latent Period (pp. 18–20) Predict Question 1: No, changes to the stimulus intensity will not change the duration of the latent period. The latent period is a chemical event initiated by the stimulus regardless of its intensity. ZAO Ch 02-1 Chart 1: Latent Period Results Voltage Active force (g) Latent period (msec) 0.0 0.00 3.0 1.04 4.0 1.32 3.20* 6.0 1.65 3.20* 8.0 1.81 3.20* 10.0 1.82 3.20* *- -Students use a visual ruler to determine the latent period. A student who enters 2.80 msec as the latent period likely understood how to correctly measure the latent period (the data points at 2.80 msec and 3.20 msec look very similar in the software). Activity Questions: 1. A graph similar to the tracing generated in the simulation. See figure 2.3 in the simulation for comparison. 2. The events of the latent period include the events of excitation contraction coupling, most notably the release of calcium from the sarcoplasmic reticulum. Activity 2: The Effect of Stimulus Voltage on Skeletal Muscle Contraction (pp. 20–22) 6. 0.8 volts Predict Question 1: The active force will first increase and then plateau at some maximal value as the stimulus voltage increases. 12. 8.5 volts Chart 2: Effect of Stimulus Voltage on Skeletal Muscle Contraction Ch 02-2 Voltage Active force (g) Voltage Active force (g) 0.0 0.00 5.0 1.51 0.2 0.00 5.5 1.59 0.8 0.02 6.0 1.65 1.0 0.15 6.5 1.70 1.5 0.43 7.0 1.74 2.0 0.66 7.5 1.78 2.5 0.87 8.0 1.81 ZAO 3.0 1.04 8.5 1.82 3.5 1.19 9.0 1.82 4.0 1.32 9.5 1.82 4.5 1.42 10.0 1.82 Activity Questions: 1. The active force produced by the muscle increased as the stimulus voltage was increased. 2. In the body this is achieved by motor unit recruitment. More muscle fibers are recruited to increase the force generated. Activity 3: The Effect of Stimulus Frequency on Skeletal Muscle Contraction (pp. 22–23) Predict Question 1: As the stimulus frequency increases, the muscle force generated by each successive stimulus will increase. There will be a limit to this increase. Predict Question 2: The stimulus frequency will need to increase. Chart 3: Effect of Stimulus Frequency on Skeletal Muscle Contraction Voltage Stimulus Active force (g) 8.5 Single 1.83 8.5 Multiple Variable, ≤2.42 8.5 Multiple Variable, ≥2.42 8.5 Multiple Variable, >2.42 and <5.20 10 Multiple Variable, >2.42 and <5.20 8.5 Multiple Variable, ≥5.20 Activity Questions: 1. Treppe is known as the staircase effect because the tracing looks like a staircase, with each subsequent wave higher than the previous wave. 2. More force is generated by the muscle with each successive twitch, thought to be due to increased availability of calcium. 3. When you increase the frequency of stimulation, the amount of force generated increases. 4. Wave summation occurs in the body when muscle fibers are stimulated before they have had a chance to completely relax. ZAO Ch 02-3 Activity 4: Tetanus in Isolated Skeletal Muscle (pp. 24–25) Predict Question 1: As the stimulus frequency increases, the muscle force generated by each successive stimulus will increase. There will be a limit to this increase. Chart 4: Tetanus in Isolated Skeletal Muscle Stimuli/second Active force (g) 50 5.12 130 5.88 140 5.91 142 5.94 144 5.94 146 5.95 148 5.95 150 5.95 Activity Questions: 1. A summation of force is occurring at a high frequency of stimulation to produce smooth muscle contraction. 2. “Lockjaw” is a pathological tetanus. Tetanus boosters are vaccines to prevent the development of tetanus, the disease. Activity 5: Fatigue in Isolated Skeletal Muscle (pp. 25–26) Predict Question 1: The length of the rest period will proportionately increase the length of time for sustained muscle tension. Chart 5: Fatigue Results Rest period (sec) Active force (g) Sustained maximal force (sec) 0 5.86 10 0 5.86 10 Variable, 8–12 5.86 Variable, 0.2–1.8 Variable, 8–12 5.86 Variable, 4.2–5.8 Activity Questions: 1. Fatigue is still being investigated, but it is thought to involve the buildup of lactic acid, ADP, Ch 02-4 ZAO and inorganic phosphate, and possibly oxygen debt. 2. They best way to delay the onset of fatigue with intense exercise is to schedule brief periods of rest to allow muscle recovery. Activity 6: The Skeletal Muscle Length-Tension Relationship (pp. 26–28) Predict Question 1: Total force can increase or decrease depending upon the starting resting length. Chart 6: Skeletal Muscle Length-Tension Relationship Length (mm) Active force (g) Passive force (g) Total force (g) 75 1.82 0.00 1.82 70 1.75 0.00 1.75 65 1.55 0.00 1.55 60 1.21 0.00 1.21 55 0.73 0.00 0.73 50 0.11 0.00 0.11 80 1.75 0.02 1.77 90 1.21 0.25 1.46 100 0.11 1.75 1.86 Activity Questions: 1. Changes in the resting length of the sarcomere directly affect the amount of passive, active and total force that results as described by the length-tension relationship. 2. The dip in the total force curve is due to the fact that at a very short muscle length, there is too much overlap to generate a significant amount of active force. Additionally, there is no passive force at this muscle length. Activity 7: Isotonic Contractions and the Load-Velocity Relationship (pp. 28–29) Predict Question 1: The latent period will increase, the shortening velocity will decrease, the distance will decrease, and the contraction duration will decrease. Chart 7: Isotonic Contraction Results Weight (g) Velocity (cm/sec) Twitch duration (msec) Distance lifted (mm) 0.5 0.100 78.00 4.0 1.0 0.057 49.00 2.0 1.5 0.022 30.00 0.5 ZAO Ch 02-5 2.0 0.00 0.00 0.0 Activity Questions: 1. As the weight of the load increases, the initial velocity to move the weight decreases. 2. This is because a heavier weight will have a slower velocity for the repetitions, so it will take you longer to repeat the same number of repetitions of a heavier weight. 2 REVIEW SHEET Exercise Skeletal Muscle Physiology NAME _______________________ LAB TIME/DATE _____________ ACTIVITY 1 The Muscle Twitch and the Latent Period 1. Define the terms skeletal muscle fiber, motor unit, skeletal muscle twitch, electrical stimulus, and latent period. See definitions provided in the Introduction. 2. What is the role of acetylcholine in a skeletal muscle contraction? Acetylcholine binds to receptors in the motor end plate, initiating a change in ion permeability that results in the end-plate potential. 3. Describe the process of excitation-contraction coupling in skeletal muscle fibers. Excitation-contraction coupling is the release of calcium which binds to troponin, removing the blocking action of tropomyosin so that myosin can bind to actin. 4. Describe the three phases of a skeletal muscle twitch. Latent period is the time preparing for contraction. Contraction is when muscle tension peaks. The relaxation period is at the end of muscle contraction. 5. Does the duration of the latent period change with different stimulus voltages? How well did the results compare with your prediction? The latent period did not change with changes in stimulus voltage. 6. At the threshold stimulus, do sodium ions start to move into or out of the cell to bring about the membrane depolarization? Sodium would move into the cell to bring about membrane depolarization. ACTIVITY 2 The Effect of Stimulus Voltage on Skeletal Muscle Contraction 1. Describe the effect of increasing stimulus voltage on isolated skeletal muscle. Specifically, what happened to the muscle force generated with stronger electrical stimulations and why did this change occur? How well did the results compare with your prediction? The active force increased as predicted to the point in which it reached a plateau and was no longer able to increase. 2. How is this change in whole-muscle force achieved in vivo? This is achieved by the recruitment of more muscle fibers over time. 3. What happened in the isolated skeletal muscle when the maximal voltage was applied? All of the muscle fibers have been recruited and so the maximal force has been achieved. Ch 02-6 ZAO ACTIVITY 3 The Effect of Stimulus Frequency on Skeletal Muscle Contraction 1. What is the difference between stimulus intensity and stimulus frequency? The stimulus intensity is the electrical changes that relate to the action potential. The frequency is the number of action potentials per minute. 2. In this experiment you observed the effect of stimulating the isolated skeletal muscle multiple times in a short period with complete relaxation between the stimuli. Describe the force of contraction with each subsequent stimulus. Are these results called treppe or wave summation? With complete relaxation, it would be treppe. This is the staircase effect, where you see an increase in the force/tension produced. 3. How did the frequency of stimulation affect the amount of force generated by the isolated skeletal muscle when the frequency of stimulation was increased such that the muscle twitches did not fully relax between subsequent stimuli? Are these results called treppe or wave summation? How well did the results compare with your prediction? The voltage needed to increase because the tension wasn’t great enough at the lower voltage. This is consistent with wave summation. 4. To achieve an active force of 5.2 g, did you have to increase the stimulus voltage above 8.5 volts? If not, how did you achieve an active force of 5.2 g? How well did the results compare with your prediction? Yes, it was necessary to increase the voltage above 8.5 volts to achieve the active force of 5.2 grams. 5. Compare and contrast frequency-dependent wave summation with motor unit recruitment (previously observed by increasing the stimulus voltage). How are they similar? How was each achieved in the experiment? Explain how each is achieved in vivo. Frequency-dependent wave summation is dependent upon stimulation by the nervous system. The motor recruitment depends upon the number of motor fibers available. ACTIVITY 4 Tetanus in Isolated Skeletal Muscle 1. Describe how increasing the stimulus frequency affected the force developed by the isolated whole skeletal muscle in this activity. How well did the results compare with your prediction? The force developed increases as the stimulus frequency increases – to a point. 2. Indicate what type of force was developed by the isolated skeletal muscle in this activity at the following stimulus frequencies: at 50 stimuli/sec, at 140 stimuli/sec, and above 146 stimuli/sec. At 50 stimuli/sec: 5.12g. At 140 stimuli/sec: 5.91g. Above 146 stimuli/sec: 5.95g 3. Beyond what stimulus frequency is there no further increase in the peak force? What is the muscle tension called at this frequency? After 146 stimuli/sec there is no further increase in force. This is the maximal tetanic tension. ACTIVITY 5 Fatigue in Isolated Skeletal Muscle 1. When a skeletal muscle fatigues, what happens to the contractile force over time? When skeletal muscle fatigues, the contractile force decreases over time. 2. What are some proposed causes of skeletal muscle fatigue? The buildup of lactic acid, ADP and inorganic phosphate are thought to be involved in muscle fatigue. 3. Turning the stimulator off allows a small measure of muscle recovery. Thus, the muscle will produce more force for a longer time period if the stimulator is briefly turned off than if the stimuli were allowed to continue without interruption. Explain why this might occur. How well did the results compare with your prediction? When you increase the rest periods, you see an increase in the muscle tension produced. ZAO Ch 02-7 4. List a few ways that humans could delay the onset of fatigue when they are vigorously using their skeletal muscles. They could periodically rest during vigorous exercise. ACTIVITY 6 The Skeletal Muscle Length-Tension Relationship 1. What happens to the amount of total force the muscle generates during the stimulated twitch? How well did the results compare with your prediction? Total force can increase or decrease depending upon the starting resting length. This is due to the length-tension relationship of the sarcomere. 2. What is the key variable in an isometric contraction of a skeletal muscle? The length-tension relationship. The passive force is important in determining the active force produced. 3. Based on the unique arrangement of myosin and actin in skeletal muscle sarcomeres, explain why active force varies with changes in the muscle’s resting length. The active forces vary with the number of crossbridges formed, which changes with the resting length of the muscle. 4. What skeletal muscle lengths generated passive force? (Provide a range.) The muscle lengths from 80-100 mm generated passive force. 5. If you were curling a 7-kg dumbbell, when would your bicep muscles be contracting isometrically? No, it would be changing in length, so this would not be isometric contraction. ACTIVITY 7 Isotonic Contractions and the Load-Velocity Relationship 1. If you were using your bicep muscles to curl a 7-kg dumbbell, when would your muscles be contracting isotonically? Yes, because your muscle is changing in length. 2. Explain why the latent period became longer as the load became heavier in the experiment. How well did the results compare with your prediction? The latent period became longer because it takes more time to generate the force required. 3. Explain why the shortening velocity became slower as the load became heavier in this experiment. How well did the results compare with your prediction? It takes more time to generate the force required to lift the heavier load. 4. Describe how the shortening distance changed as the load became heavier in this experiment. How well did the results compare with your prediction? The shortening distance decreased with the heavier load. 5. Explain why it would take you longer to perform 10 repetitions lifting a 10-kg weight than it would to perform the same number of repetitions with a 5-kg weight. The velocity of shortening decreases with a heavier load, so the repetitions will take longer with a 10 kg weight. 6. Describe what would happen in the following experiment: A 2.5-g weight is attached to the end of the isolated whole skeletal muscle used in these experiments. Simultaneously, the muscle is maximally stimulated by 8.5 volts and the platform supporting the weight is removed. Will the muscle generate force? Will the muscle change length? What is the name for this type of contraction? The muscle will still generate force and change length. The type of contraction is isotonic. Ch 02-8 ZAO