Excitation Contraction Coupling

Nandini Goyal
 Session Overview
• Structure of Neuromuscular Junction
• Neuromuscular Transmission
• Excitation Contraction Coupling
• Structure of Muscle Proteins
• Sarcomere – Definition and Structure
• Process of Muscle Contraction
• Summary
 Neuromuscular Transmission

• The junction between a motor neuron and a muscle fiber is called

neuromuscular junction.
• It is also known as myoneural junction or motor end plate.
• Action potential from the neuron is transmitted to the muscle
through this junction.
• Physiological anatomy of neuromuscular junction or myoneural
junction or motor end plate :
1. Axon Terminal (Presynaptic)
2. Synaptic Cleft
3. Post Junctional Membrane (Postsynaptic)
 Axon Terminal
• The axon of the motor neuron divides extensively into several
branches called axon terminal. Each terminal forms a junction with
single skeletal muscle fiber midway along its length.
• The motor neuron including its axon terminals and muscle fibers is
called a motor unit.
• Each axon terminal is dilated at its end to form a knobby structure
called synaptic knob, which contains a large number of mitochondria
neurotransmitter vesicles and active zones.
 Synaptic Cleft

• It is a space 40-100 nm wide, between the axon terminal and post

junctional membrane.
• It is filled with ECF consisting of collagen fibers and glycoproteins.
• It contains the enzyme Acetylcholine Esterase (AChE).
 Post Junctional Membrane
• It is the part of sarcolemma that lies directly under the axon terminal.
• The area of post junctional membrane is thrown into several folds
called as junctional folds which increase the area of the membrane.
• It contains numerous ACh receptors (nicotinic) which are
concentrated at the junctional folds. It also contains voltage gated Na+
channels.
 Mechanism of Neuromuscular Transmission
• Action potential arrives at the axon terminal causing its
depolarization.
• This causes the opening of voltage gated Ca+ channels.
• Ca+ influx leads to the migration of neurotransmitter vesicles towards
the axon terminal membrane.
• The vesicles fuse with the membrane leading to their exocytosis, thus
releasing Ach in the synaptic cleft. This is facilitated by synaptobrevin,
a vesicular membrane protein with syntaxin, a neuronal membrane
protein.
• Their released ACh molecules diffuse through the synaptic cleft and
bind to the ACh receptors present on the post junctional membrane.
• Binding of ACh to ACh receptors allows Na+ influx due to the
opening of ACh gated Na+ channels producing local depolarization
known as End Plate Potential (EPP).
• EPP leads to opening of voltage gated Na+ channels present over
the sarcolemma causing the generation of action potential.
• The action potential spreads along the sarcolemma on either
sides, dips into the muscle fiber through the T-tubules and
initiates contraction.
 Excitation Contraction Coupling
• The action potential on the surface of cell membrane
triggers the contraction of the muscle fiber using Ca++ ions.
Thus, the phenomenon of pairing electrical event with
mechanical event is called excitation contraction coupling.
• Sarcotubular System - The sarcolemma along with the
sarcoplasmic reticulum forms a highly specialized system
called as sarcotubular system. This plays an important role
in the depolarization inside muscle fiber. It consists of :
• T-tubules (Transverse) - It is formed by the invagination of the
sarcolemma in the muscle fiber at the region of A-I junction. Their
lumen contains ECF. They are responsible for carrying the action
potential into the muscle fiber thus activating all sarcomeres.
• L-tubules (Longitudinal) - These are modified ER. They are present on
either sides of the T-tubule and are dilated to form terminal cisternae.
Thus a T-tubule with two terminal cisternae in close proximity
constitute a triad. Thus there are two triads in each sarcomere.
• In cardiac muscle it forms a diad due to the presence of the one
cistern.
• There are certain processes called as junctional feet between the T-
tubules and terminal cisternae which involve two integral membrane
proteins i.e. ,
• 1) In the T-tubule membrane DHP (Dihydropyridine) modified voltage
sensitive L-type Ca++ channels - primarily acts as a voltage sensor
rather than a Ca++ channel.
• 2) In the terminal cisternae membrane Ryanodine receptors mainly
act as Ca++ release channels.
Process of Excitation Contraction Coupling
• When action potential reaches the T-tubule it activates the voltage
sensitive Ca++ channels i.e. DHP receptors located in the T-tubule
membrane.
• This in turn triggers the opening of Ca++ release channels i.e.
ryanodine receptors located in the terminal cisternae membrane.
• (The DHP when activated by the depolarization of T-tubules
undergoes conformational changes which result in opening of the
ryanodine).
• Thus Ca++ ions diffuse into the cytoplasm and get attached to
troponin C and start a chain of events which produce contraction.
• This forms the basis of excitation contraction coupling.
 Molecular Basis Of Skeletal Muscle
Contraction
• A typical skeletal muscle contains many muscle fascicles each of
which contains a large number of muscle fibers arranged parallel to
each other.
• Connective tissue layers around :
1. Muscle – Epimysium
2. Individual fascicle – Perimysium
3. Muscle fiber – Endomysium
• The most striking feature of skeletal muscle is the presence of
striations due to alternate dark (A band) and light (I band) bands with
respect to their behavior to polarized light.
 Muscle Proteins
• Each myofibril is made of sarcomere which is the structural and
functional unit of muscle fiber that contains different muscle proteins
i.e. ;
A. Contractile proteins – Actin and Myosin making the thick and thin
filaments.
B. Regulatory proteins – Troponin and Tropomyosin.
C. Attachment proteins - Titin, Nebulin, Desmin, Dystrophin.
 Myosin
• The thick filaments are polymers of myosin II. (MW – 4,80,000)
• Each myosin filament is composed of two globular heads, neck (hinge region)
and a long tail.
 MYOSIN HEAD - The myosin head contains two binding sites, one for ATP
and other for actin. The ATP binding site functions as ATPase.
MYOSIN TAIL – The tail of each myosin molecule lies along the axis of the
thick filament and the two globular heads extend out to the sides, forming
cross bridges.
MYOSIN HINGE – In this region, the tail joins the globular head.
• The myosin molecules aggregate with a definite directional arrangement
such that their tail ends are directed towards the center of the thick
filaments creating a bare region in the middle consisting of myosin tails only,
while the globular heads point away from both sides of the tail.
• M line indicates the site of reversal of polarity.
 Actin
• The thin filaments are made of actin.
• Monomers of G-actin (globular) molecules (MW – 42,000) are joined
from front to back into long chains that wind about each other
forming a double stranded alpha helical filament known as F-actin
(filamentous) that forms the backbone of thin filament.
• Each actin monomer contains binding sites for myosin, tropomyosin,
troponin I.
 Tropomyosin
• Tropomyosin is a rod shaped molecule (MW – 70,000) composed of
two bands of polypeptides intertwined in an alpha helical
configuration with a length approximately equal to seven actin
monomers.
• The troponin molecule extends over the entire length of actin
filament covering myosin binding sites on the actin monomers.
• The function of tropomyosin is to interfere with the binding of myosin
head to action.
 Troponin
• Troponin is a complex of three proteins :
1. Troponin T – It binds troponin complex to tropomyosin.
2. Troponin I – It binds the troponin complex to actin. It inhibits the
binding of actin to myosin by blocking the myosin binding site on
actin.
3. Troponin C – It binds to calcium.
 Sarcomere
• Sarcomere is the structural and functional unit of a myofibril.
• It is defined as the portion of the muscle fiber between two
consecutive Z lines.
 Mechanism of Skeletal Muscle Contraction
• AF Huxley in 1957 proposed the sliding filament theory of muscle
contraction based on the changes observed in the lengths and widths
of various bands and zones of the sarcomere.
• This occurs by the sliding of thin filaments over thick filaments
towards the center of the sarcomere.
Q: WHAT HAPPENS TO THE BANDS WHEN A MUSCLE CONTRACTS?
• Width of A band remains same.
• Width of I band is reduced.
• In a fully contracted muscle, the H zone disappears.
Sliding Filament Theory or Walk Along Theory
or Ratchet Theory
• Following propagation of action potential along the sarcolemma and
down the T-tubules, Ca++ is released from the sarcoplasmic reticulum.
The Ca++ concentration rises to 10-4 M.
• Ca++ binds to troponin C and induces a conformational change in the
troponin molecule.
• This results in shifting of troponin I from the actin filament allowing
the tropomyosin to move deeper into the actin groove unmasking the
myosin binding site.
• This allows the myosin heads to interact with actin and engage in
formation of cross bridges.
• The intrinsic ATPase activity of the myosin heads breaks down
cytoplasmic ATP to ADP and Pi.
• This results in the rotation of myosin head around the hinge in such a
way that the globular head is positioned at 90 degree angle in relation
to the thick filament.
• This rotation moves the myosin head about 10 nm along the thin
filament. This resting confirmation is known as cocked state or
energized state of the myosin head.
• The myosin head then binds to actin resulting in formation of cross
bridges. Thus the formation of actomyosin complex has 2 effects :
- Pi released from the myosin head produces a conformational change
in which the myosin head bends on the hinge at an angle of about 45
degree pulling the actin filaments 10 nm towards the center of the
sarcomere. This is known as power stroke.
- The binding the dissociation of ADP from the myosin head leaving the
actomyosin complex in a rigid state.
• For detachment to occur a new molecule of ATP must bind to the
myosin head and initiate the next cycle.
• As long as the muscle is stimulated the cross bridge cycling repeats
many times dragging the thin filaments attached to successive Z lines
towards the center of sarcomere which leads to decrease in
sarcomeric length and ultimately muscle shortening.
Summary
Thankyou.