Properties of Skeletal

Muscle
Lab 2

Purpose
To see the effects of direct muscle
stimulation
To analyze the effects of stimulus intensity
and stimulus frequency on contraction
To analyze the effects of acetylcholine
(ACh) receptor blockade on muscle activity

Skeletal muscle

Skeletal muscle makes up ~40%

of body weight.
Primary function: to generate
force or movement. Responsible
for positioning and moving the
skeleton.

Muscle Fibersindividual muscle

cells
Muscle fibers
contain many
myofibrils
Myofibrilscomposed of long
proteins (actin,
myosin, and other
proteins).
These proteins are

1. Molecular basis of contraction

1 m
Sarcomeres are basic contractile unit of muscle fiber (functional
unit).

Contraction involves shortening of each

sarcomere

Structure of thin and thick filaments

Actin-myosin interactions and cross-bridge cycling

are regulated by [Ca2+]

The cross-bridge cycle in skeletal muscle

Need Ca+2
and ATP in
order to
continue with
the CBC.

Excitation-Contraction Coupling
Neuromuscular Junction:
Area where the motor
neuron synapses with the
muscle fiber
Depolarization causes the
release of Acetylcholine
(ACh) into the synaptic cleft
ACh binds to nicotinic ACh
Receptors (nAChR) causing
them to open
nAChR are ligand gated
cation channels (Do Not
confuse with muscarinic ACh
receptors)

Action potentials race down

the sarcolemma and down
into the muscle via T-Tubules
AT-tubule(ortransverse
tubule) is a deep
invagination of the
sarcolemma, which is the PM
of skeletal muscle and cardiac
muscle cells.
These invaginations allow
depolarization of the
membrane to quickly
penetrate to the interior of
the cell.

Calcium Release

Summary of molecular events in EC coupling

- Depolarization

activates voltage-gated
Ca channels (DHPR),
which open Ca release
channels (RyR) in SR
- Ca binds to troponin,
allowing actin/myosin
binding and power
stroke

Relaxation occurs when cytosolic [Ca2+]

decreases

3. Neural regulation of contraction

Motor unit consists of one
motor neuron and the
muscle fibers that it
innervates (10s - 1000s).
Motor pool consists of all
motor neurons innervating
a single muscle.
Contraction is regulated by
2 mechanisms:
a. stimulation frequency of
individual MUs
b. recruiting different
numbers of MUs.

a. Frequency of stimulation of motor unit

Force is increased by summing twitches in each muscle fiber.

b. Progressive recruitment of motor units

H principle
- In whole muscle, force

increases as additional MUs

are recruited.
- Progressive recruitment of
first small and then larger
motor units - referred to as
the size principle.
- Force is increased by
summing contractions of
fibers in each MU.

4. Muscle mechanics

Contractions that occur at constant length are called isometric

contractions (generate force but no muscle shortening)
Contractions that occur against constant load are called
isotonic contractions (generate force greater than load and
shortening occurs)

Length-Tension Relationship

Summary

1. The contractile unit of the muscle fiber is the sarcomere,

which is composed primarily of myosin and actin
filaments. Contraction results from filament sliding.
2. E-C coupling involves Ca release from the SR, which then
acts via the troponin complex to initiate cross-bridge
cycling.
3. Neural regulation of contraction is mediated by the motor
unit - frequency of stimulation of the motor unit and by
recruiting different numbers of motor units.
4. Force of contraction depends on muscle length (number of
crossbridges formed).

 Part 1: Equipment Set-up

1. Set-up computer and calibrate force
transducer
2. Isolate gastrocnemius from frog and attach to
transducer
3. Isolate sciatic nerve and connect to electrode

Notes:
Have 1 or 2 people set-up the computer
and calibrate the transducer while the
others dissect the frog.
Be very careful with the sciatic nerve. Do

Part 2: Experiment & Data Collection

Part 2(a): Effect of stimulus intensity on
Muscle Activity-Threshold & Max
You will increase the voltage used to stimulate
the sciatic nerve to find the lowest voltage that
elicits a twitch and the voltage at which the
twitch height no longer increases
You will then increase the stimulus voltage by
steps between these values to look at how
stimulus intensity effects tension

Q: What do you expect to happen to

tension as the stimulus intensity increases
and why?

 Part 2(b): Effect of stimulus frequency on

muscle activity
You will stimulate the muscle at the maximum
voltage found in part 2(a) and increase the
frequency the stimulus is given

Q: What do you expect to happen to tension

if the frequency of stimulation is increased
and why?

 Part 2(c): Effect of Tubocurare on muscle

activity
Tubocurare is a plant alkyloid that is a
competitive inhibitor of nAChR
You will look at the effect of the addition of
tubocurare on muscle contraction
Note: Remember to take control tension
measurements before adding Tubocurare

Q: What do you expect to happen to muscle

contraction after tubocurare is added and
why?

Competitive
Inhibition
ACh
ACh

nACh
R

ACh

nACh
R

Depolariza
tion

TC

TC

nACh
R

nACh
R

Depolariza
tion

Generally, you can overcome competitive inhibition by

increasing the concentration of the agonist (in our case

 Part 2(d):Effect of Direct stimulation on

muscle activity
Here you will be directly stimulating the muscle
itself to cause a contraction instead of
stimulating the sciatic nerve

Q: Do you expect the same general effect

of tubocurare on muscle tension if you
directly stimulate the muscle and why?

Notes for lab:

Remember to mark (F9) and label your
graphs as you are collecting the data
When add tobucurare press F9 at the beginning
and at the end of the injection.

Wear gloves when handling the frog, but

DO NOT touch the computer with gloved
hands
Wash tools used and place on the back
table to dry

Quizzes
Quizzes are given the first 10-15 min of lab.
Please arrive on time, no extra time will be
given if you are late.
Approximately, half of the quiz will cover
material from the previous lab and the
other half the material from the current
days lab
For example: You should be familiar with the
material & methods used, concepts behind the
lab, goals of the lab, expected results, etc
From the previous lab you should understand
the physiological concepts, material we cover
in the discussion section, interpretation of data,
etc

Before you come to lab Bring signed NPB 101L Contact Form
Read REQUIRED READING on SmartSite (will be on the
quiz)
Read Preface of lab note book, Lab 2 introduction and
methods