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Tutorial 1

PREPERED BY :
Dr. Radwa Ali Mehanna
Dr. Eman Magdy

SUPERVISION BY :
Dr/Samar El ghotny

physiology department
Definition Of Simple
Muscle Twitch
A single brief contraction followed
by a relaxation of a skeletal
muscle due to application of a
single adequate stimulus.
SO We
Need
Recording apparatus
Muscle
Stimulus
Frog
? Electrical stimulus
-As human
-cheap
?
-As nature
-No permanent damage
-Easy for application
 Recording apparatus :
The apparatus which is commonly used
in the routine laboratory experiments on
the muscle and nerve consists of :
1. Stimulation set ( delivers electrical
stimulus for the muscle nerve preparation
).
2. kymograph .
3. Myograph .
Stimulation set :
1. source of galvanic current .
2. Induction coil .
3. Connecting wires.
4. Simple key .
5. Two stimulating electrodes .
Myograp
h
Frog Dissection

Fall 2000
Jenna Hellack
 OBJECTIVE: TO RECORD SIMPLE MUSCLE
TWICH (SMT)

 LEARNING OBJECTIVES:
 By the end of this experiment student should know:
 How to pith the frog.
 How to prepare nerve and muscle for further procedure.
 How to record a graph of simple muscle twitch.
 TO PITH THE FROG
 REQUIREMENTS:

Frog and dissecting instruments.


 THEORY:
 Before using any animal for
experimental purposes it should be
made unconscious.
 To produce unconsciousness there
are two methods:
Anesthesia:
The unconsciousness may be produced
by using any anesthetic agent which may
be used orally, intravenously or by
inhalation of chloroform or ether, etc. but it
is very costly and carries its own risk.
Pithing:
The unconsciousness may be
produced
by destroying the brain and spinal cord
that is the CNS.
This prevents the frog from perceiving
any pain (the frog is clinically dead).

Pithed Frog
PREPARATION OF NERVE AND MUSCLE OF
FROG
OBJECTIVE:
The student should be able to :
•Prepare gastrocnemius sciatic preparation.
•Explain the effects on nerve and muscle if not
placed in Ringer's Solution.
REQUIREMENTS:
Frog, dissecting instruments, Ringer's
solution, beaker, thread, dissecting board,
glass rod, Common pin.
PRINCIPLE:
The principle of this experiment is to obtain
a separate independent muscle along with its
nerve supply so that:
It may be stimulated and various effects
observed
PROCEDURE:
Lay the pithed frog on the frog board.

 Cut the skin transversely all around the trunk and


remove the skin from the two hind Limbs.

Open the abdominal cavity, remove the viscera, a


white, shining nerve may be seen emerging from the
last 3 vertebrae.

Give a transverse cut just over the last 3 vertebrae


and cut the two hind limbs into two pieces by giving
a longitudinal cut very carefully so that the nerve is
not damaged.
Remove all the muscles of the thigh
completely and also cut the femur just near
the knee joint so that a small portion of
femur remains attached with the knee joint.

 The sciatic nerve is completely exposed


and only a portion of the last 3 vertebrae
remain attached with sciatic nerve.
Free the sciatic nerve from surrounding tissue
Expose the nerve in the thigh where it lies
deep between the large thigh muscles
Now separate the muscle with the help of a
glass rod, tie a thread at Achilles tendon and
cut it about 1 cm away from the knot.

Now cut the tibia just near the knee joint so


that a small portion of it remains attached
with the knee joint.

This is Nerve Muscle Preparation or


Gastrocnemious Sciatic Preparation.
Gastrocnemius m with its nerve
sciatic nerve
What is the meaning of
after loaded muscle and
freeloaded muscle?
 To understand the meaning of free
loaded and after loaded muscle we
must return to the myograph ,a part
of the recording apparatus .
 The myograph board has a metalic
projection which carries a lever .
 The lever has two arms :

a. Horizontal arm to which is fitted a


pointer with a writing point at its end

b.Vertical arm to which is attached a


thread ligated in the tendon of the
muscle ,when the preparation is
mounted on the myograph .
Horizontal arm
 the lever is also
provided with a
screw called
after loading
screw . This
screw can be
moved up and
down to make
the muscle free
loaded or after
loaded .
After loaded muscle
If the weight ( lever) attached to the tendon
of a muscle is supported by the after
loading screw during relaxation and does
not pull the muscle except when the
muscle start to shorten
i.e the muscle carries no weight at the start
of contraction ( unstretched)

The muscle is loaded after start of


contraction
Free loaded muscle
When the load (weight of the lever) pulls the
muscle during relaxation and increases its
length i.e Muscle is stretched ( this is done
by opening the after loading screw)
Free loaded muscle is capable of giving
stronger contraction provided that the
weight does not over stretch the muscle
Freeloaded and afterloaded
muscle
Tension varies with the starting length of the
sacromere
Method Of Stimulation

Direct stimulation Indirect stimulation

Direct stimulation of the Stimulation of the nerve


muscle which supply the muscle
Which is better ?

Direct stimulation Indirect stimulation

Supply to all muscle fiber


Same as nature of our body
Recording Of Simple
Muscle Twitch
Phases of SMT
Latent phase : the time that passes between
the application of the stimulus to start of
contraction
Contraction phase :the period which the
muscle shorten in length
Relaxation phase : the period during the
muscle restores its initial length
Duration of the SMT
Total twitch time(TTT) = ,1 sec

Latent phase : ,01 sec

Contraction phase : ,04 sec

Relaxation phase : ,05 sec


Causes Of Latent
Phase
1- Action potential in the
nerve
1
2- Chemical transmission

2
3- Action potential in the
muscle
3
Chemical Transmission
Types Of Contraction
Isotonic Contraction Isometric Contraction
 Same tension  Tension Increase
 Muscle  Same Length
shortening  No Work Is Done
 Work is done  Energy Lost as
 Energy ¼ work Heat
& ¾ lost as heat
Isotonic and Isometric
Contractions
Factor Affecting SMT

1- Warming
2- Cooling
3- Fatigue
Warming
 warm saline (physiological
saline of frog ,69% Nacl)
30-38C.
 Duration decrease ,
amplitude increase.
 Mechnism :↑ metabolic warming

chemical reaction &


↓viscosity of the muscle
 Over heating produce
SMT
irreversible coagulation of
its protein ( heat rigor)
Cooling
 Cold saline (physiological
saline of frog ,69% Nacl) 0-
4C.
 Duration increase , amplitude
decrease
 Mechnism :↓ metabolic
chemical reaction & SMT

↑viscosity of the muscle


 Excessive cooling or freezing COOLING

lead to loss of vitality


&irritability
Fatigue
 By Repeated successive stimulation .
 Duration increase , amplitude decrease.
 Mechanism : depletion of energy stores &
accumulation of metabolites as lactic acid
 The first site of fatigue motor end plate

SMT

fatigue
Motor End Plate
Multiple Stimulation
 The effect of applying multiple successive
stimuli to a muscle depends upon the
frequency of these stimuli ( duration
between them),which can be either:
 Low frequent stimuli
 Moderate frequent stimuli
 High frequent stimuli
Low Frequent
Stimulation
 In which each stimulus fall after the
complete muscle relaxation of the previous
stimulus.
 So it results in the production of successive
simple muscle twitches ( contraction then
relaxation) which are called Separate
Twitches
Moderate frequent
stimuli
 In which each stimulus fall during muscle
relaxation of the previous stimulus
 So it results in the production of
successive contractions each followed by
incomplete relaxation which are called
Clonus or
Incomplete Tetanus
Clonus or Incomplete Tetanus

Stimulus applied Stimulus applied

Muscle fiber was not able


to relax so tension
increased
High frequent stimuli
 Inwhich each stimulus fall during muscle
contraction of the previous stimulus.
 So it results in the production of successive
contractions ( contraction on top of
contraction ) which is called Complete
Tetanus
Complete Tetanus
Stimuli

When action potentials come VERY RAPIDLY


muscle fiber CANNOT relax result in
contraction on top of contraction . i.e
sustained contraction
Treppe or staircase
phenomenon
 This phenomenon states that with
successive stimulation the force of
contraction increases although the
intensity is constant
 It occurs due to successive stimulation of
the muscle whatever is the frequency of
stimulation
Causes of treppe
phenomenon
 In low frequent stimuli:
 It is due to increased heat production

Heat

Muscle viscosity which


acts as resistance to
contraction
 In Moderate frequent stimuli:
 It
is due to increased heat production
 Accumulation of Ca++ ions
As the stimulus reach the muscle while relaxing
( ie relaxation is not complete) the Ca2+ is not
completely pumped back to SR
So the stimulus adds Ca2+ to the already
present from the previous twitch and so on
More ca2+ increase force of
contraction
Treppe in Moderate frequent stimuli
 In High frequent stimuli:
 Itis due to increased heat production
 Accumulation of Ca++ ions
 Possibility of falling of the stimulus in the
supra normal phase of excitability
Excitability Of Skeletal
Muscle
 Excitability
Ability to receive and respond to a stimulus
Relation between
excitability and
mechanical changes
Tetanus never occurs in
the cardiac muscle
Factors Affecting
Clonus
 Warming
 Cooling
 Fatigue
Warming

Muscle warming coverts the clonus into separate


twitches as:
Warming accelerates the chemical reactions ,thus all
phases of the muscle twitch are shortened
The stimulus will fall at the end of relaxation instead of
falling during relaxation
Cooling

Muscle cooling coverts the clonus into tetanus:


cooling decelerates the chemical reactions ,thus all
phases of the muscle twitch are prolonged
The stimulus will fall during contraction instead of falling
during relaxation
Fatigue
Prolonged stimulation of a muscle leads
to muscle fatigue. It is due to:

 Depletion of ATP, glycogen & creatine


phosphate ( energy stores)

 Accumulation of metabolites (lactic


acid) and increased acidity.
Fatigue

Tetanus
fatigue

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