Sumihar Pasaribu

Departemen Biokimia Fakultas kedokteran Universitas Methodist Indonesia

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Muscle Contraction

Classification of the Muscle

According to the structure: Striated Muscle, Smooth Muscle According to the nerve innervation: Voluntary Muscle, Involuntary Muscle According to the Function: Skeletal Muscle, Cardiac Contraction, Smooth Muscle
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Skeletal Muscle

Cardiac Muscle

Smooth Muscle

I Signal Transmission Through the Neuromuscular Junction

Skeletal Muscle Innervation

Illustration of the Neuromuscular Junction (NMJ)

New Ion Channel Players

Voltage-gated Ca2+ channel
in presynaptic nerve terminal mediates neurotransmitter release

Nicotinic Acetylcholine Receptor Channel

in muscle neuromuscular junction (postsynaptic membrane, or end plate) mediates electrical transmission from nerve to muscle

Nerve Terminal

2+ Ca

channels

Structurally similar to Na+ channels Functionally similar to Na+ channels except

activation occurs at more positive potentials activation and inactivation much slower than Na+ channels

Neuromuscular Transmission

Axon Axon Terminal

Skeletal Muscle
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Depolarization of terminal opens Ca channels Nerve action +

potential invades + axon terminal

+

Neuromuscular Transmission:
+

Step by Step + +

-++

Look - + here + -

-+

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ACh is released and diffuses across synaptic cleft. ACh ACh ACh
Ca2+

Ca2+ induces fusion of vesicles with nerve terminal membrane.

Ca2+

Na+

Binding of ACh opens channel pore that is permeable to Na+ and K+.
Na+ Na+ Na+ K+ Na+

ACh binds to its + Na receptor on the K+ postsynaptic membrane +

K Na+

Na+

Na+ K+

ACh

K+

Outside

Muscle membrane
Na+ K+ Na+ K+ K+ Na+ Na+ K+ K+ K+ K+ Na+
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Inside

K+

End Plate Potential (EPP)

Muscle Membrane Voltage (mV) The movement of Na+ and K+ depolarizes muscle membrane potential (EPP)
VNa 0 EPP
Threshold

Presynaptic terminal

-90 mV
VK Presynaptic AP Time (msec)

Outside Inside

Muscle membrane ACh Receptor Channels Voltage-gated Na Channels

Inward Rectifier K Channels 13

ACh
Choline

Meanwhile ...
so the channel closes

ACh ACh
Choline

5.Choline is taken up into nerve terminal

3.Choline resynthesized into AChACh and repackaged into vesicle ACh

2.ACh is hydrolyze AChE into Choline and acetate

Acetate

1.ACh unbinds from its receptor Outside Muscle membrane

Inside
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Structural Reality

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Neuromuscular Transmission
Properties of neuromuscular junction 1:1 transmission: A chemical transmission which is designed to assure that every presynaptic action potential results in a postsynaptic one An unidirectional process Has a time delay. 20nm/0.5-1ms Is easily affect by drugs and some factors The NMJ is a site of considerable clinical importance
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Clinical Chemistry
Related compounds Suberyldicholine is are a useful in the neuroscience synthetic neuromuscular research agonist. Tubocurarine and other, related compounds are used to paralyze muscles during surgery. So tubocurarine is a neuromuscular blocking agent. Tubocurarine competes Tubocurarine is the with ACh for binding primary paralytic to receptor- but does ingredient in curare. not open the pore. 17

Ach is the natural agonist at the neuromuscular junction.

chol and related ounds are used lly for GI disorders, ma, salivary malfunction, etc.

bachol is a hetic agonist hydrolyzed by ylcholinesterase.

Anticholinesterase Agents
Anticholinesterase (anti-ChE) agents inhibit acetylcholinesterase
prolong excitation at the NMJ

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Anticholinesterase Agents
1. Normal:

ACh
AChE

Choline + Acetate

2. With anti - AchE:

ACh
anti - AChE

Choline + Acetate

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Uses of anti-ChE agents

Clinical applications (Neostigmine, Physostigmine) Insecticides (organophosphate ) Nerve gas (e.g. Sarin )

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Sarin and Aum Shinrikyo

Aum Shinrikyo is a Japanese religious cult obsessed with the apocalypse . The previously obscure group became infamous in 1995 when some of its members released deadly sarin nerve gas into the Tokyo subway system, killing 12 people and sending more than 5,000 others to hospitals.
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Sarin
Sarin, which comes in both liquid and gas forms, is a highly toxic and volatile nerve agent developed by Nazi scientists in Germany in the 1930s. Chemical weapons experts say that sarin gas is 500 times more toxic than cyanide gas.
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NMJ Diseases
Myasthenia Gravis
Autoimmunity to ACh receptor Fewer functional ACh receptors Low safety factor for NM transmission

Lambert-Eaton syndrome
Autoimmunity directed against Ca2 channels Reduced ACh release Low safety factor for NM transmission
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II Microstructure of Skeletal Muscle

Skeletal Muscle
Human body contains over 400 skeletal muscles
40-50% of total body weight

Functions of skeletal muscle

Force production for locomotion and breathing Force production for postural support Heat production during cold stress
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 Fascicles: bundles, CT(connective tissue) covering on each one Muscle fibers: muscle cells

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Structure of Skeletal Muscle: Microstructure

Sarcolemma
Transverse (T) tubule Longitudinal tubule (Sarcoplasmic reticulum, SR

Myofibrils
Actin (thin filament)
Troponin Tropomyosin

Myosin (thick filament)

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Within the sarcoplasm

Triad

Transverse tubules
Sarcoplasmic reticulum -Storage sites for calcium Terminal cisternae - Storage sites for calcium
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Microstructure of Skeletal Muscle (myofibril)

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Sarcomeres
Sarcomere : bundle of alternating thick and thin filaments Sarcomeres join end to end to form myofibrils
Thousands per fiber, depending on length of muscle

Alternating thick and thin filaments create appearance of striations

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 Myosin head is hinged Myosin Bends and straightens during contraction

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Thick filaments (myosin)

Bundle of myosin proteins shaped like doubleheaded golf clubs Myosin heads have two binding sites
Actin binding site forms cross bridge Nucleotide binding site binds ATP (Myosin ATPase)

Hydrolysis of ATP provides energy to generate power stroke

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Thin filaments

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Thin filaments (actin)

Backbone: two strands of polymerized globular actin fibrous actin
Each actin has myosin binding site

Troponin
Binds Ca2+; regulates muscle contraction

Tropomyosin
Lies in groove of actin helix Blocks myosin binding sites in absence of Ca2+

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 Thick filament: Myosin (head and tail) Thin filament: Actin, Tropomyosin, Troponin (calcium binding site)

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III Molecular Mechanism of Muscular Contraction

The sliding filament model
Muscle shortening is due to movement of the actin filament over the myosin filament Reduces the distance between Z-lines

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The Sliding Filament Model of Muscle Contraction

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Changes in the appearance of a Sarcomere during the Contraction of a Skeletal Muscle Fiber

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Cross-Bridge Formation in Muscle Contraction

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Energy for Muscle Contraction

ATP is required for muscle contraction
Myosin ATPase breaks down ATP as fiber contracts

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Nerve Activation of Individual Muscle Cells (cont.)

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Excitation/contraction coupling
Action potential along T-tubule causes release of calcium from cisternae of TRIAD Cross-bridge cycle

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Begin cycle with myosin already bound to actin

1. Myosin heads form cross bridges

Myosin head is tightly bound to actin in rigor state Nothing bound to nucleotide binding site

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2. ATP binds to myosin

Myosin changes conformation, releases actin

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3. ATP hydrolysis
ATP is broken down into:
ADP + Pi (inorganic phosphate)

Both ADP and Pi remain bound to myosin

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4. Myosin head changes conformation

Myosin head rotates and binds to new actin molecule Myosin is in high energy configuration
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5. Power stroke
Release of Pi from myosin releases head from high energy state Head pushes on actin filament and causes sliding Myosin head splits ATP and bends toward H zone. This is Power stroke.
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6. Release of ADP
Myosin head is again tightly bound to actin in rigor state Ready to repeat cycle

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THE CROSS-BRIDGE CYCLE Relaxed state Crossbridge energised

A + M l ADP l Pi

Crossbridge attachment

Ca2+ present
A M l ATP AlMlADPlPi

Crossbridge detachment ATP

Al M

Tension develops ADP + Pi

A, Actin; M, Myosin

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Cross Bridge Cycle

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Rigor mortis
Myosin cannot release actin until a new ATP molecule binds Run out of ATP at death, cross-bridges never release

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Many contractile cycles occur asynchronously during a single muscle contraction

Need steady supply of ATP!

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Regulation of Contraction
Tropomyosin blocks myosin binding in absence of Ca2+ Low intracellular Ca2+ when muscle is relaxed

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+2 Ca binds

to troponin during contraction

Troponin-Ca+2 pulls tropomyosin, unblocking myosin-binding sites Myosin-actin cross-bridge cycle can now occur
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How does

2+ Ca

get into cell?

Action potential releases intracellular Ca2+ from sarcoplasmic reticulum (SR)

SR is modified endoplasmic reticulum Membrane contains Ca2+ pumps to actively transport Ca2+ into SR Maintains high Ca2+ in SR, low Ca2+ in cytoplasm
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 How does the AP trigger contraction?

The action potential triggers contraction

This question has the beginning (AP) and the end (contraction) but it misses lots of things in the middle!
We should ask:
how does the AP cause release of Ca from the SR, so leading to an increase in [Ca]i? how does an increase in [Ca]i cause contraction?

Structures involved in EC coupling

Contractile proteins in striated muscle are organised into sarcomeres T-tubules and sarcoplasmic reticulum are organised so that Ca release is directed toward the regulatory (Ca binding) proteins

A band
Z disc

I band
(actin)

(myosin)

Z disc

M line
sarcoplasmic reticulum

Z disc

The association of a t-tubule with SR on either side is often called a triad (tri meaning three)

t-tubules
Triad

junctional feet

Structures involved in EC coupling - Skeletal Muscle T-tubule sarcolemma out in sarcoplasmic reticulum

voltage sensor?

junction foot

Ca2+ Controls Contraction

Ca2+ Channels and Pumps Release of Ca2+ from the SR triggers contraction Reuptake of Ca2+ into SR relaxes muscle So how is calcium released in response to nerve impulses? Answer has come from studies of antagonist molecules that block Ca2+ channel activity
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Dihydropyridine Receptor
In t-tubules of heart and skeletal muscle Nifedipine and other DHP-like molecules bind to the "DHP receptor" in t-tubules In heart, DHP receptor is a voltage-gated Ca2+ channel In skeletal muscle, DHP receptor is apparently a voltage-sensing protein and probably undergoes voltage-dependent conformational changes
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Ryanodine Receptor
The "foot structure" in terminal cisternae of SR Foot structure is a Ca2+ channel of unusual design Conformation change or Ca2+ -channel activity of DHP receptor apparently gates the ryanodine receptor, opening and closing Ca2+ channels Many details are yet to be elucidated!
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Skeletal muscle
The AP: moves down the t-tubule voltage change detected by DHP receptors
DHP receptor is essentially a voltage-gated Ca channel

T-tubule sarcolemma out in sarcoplasmic reticulum

is communicated to the ryanodine receptor

which opens to allow Ca out of SR

activates contraction

voltage sensor (DHP receptor)

junctional foot (ryanodine receptor)

Cardiac muscle
The AP: moves down the t-tubule
T-tubule sarcolemma out in sarcoplasmic reticulum

voltage change detected by DHP receptors (Ca channels) which opens to allow small amount of (trigger) Ca into the fibre
Ca binds to ryanodine receptors which open to release a large amount of (activator) Ca (CACR) Thus, calcium, not voltage, appears to trigger Ca release in Cardiac muscle!

voltage sensor junctional foot & Ca channel (DHP receptor) (ryanodine receptor)

The Answers!
Skeletal
The trigger for SR release appears to be voltage (Voltage Activated Calcium Release- VACR) The t-tubule membrane has a voltage sensor (DHP receptor) The ryanodine receptor is the SR Ca release channel Ca2+ release is proportional to membrane voltage

Cardiac
The trigger for SR release appears to be calcium (Calcium Activated Calcium Release - CACR) The t-tubule membrane has a Ca2+ channel (DHP receptor) The ryanodine receptor is the SR Ca release channel

The ryanodine receptor is Ca-gated & Ca release is proportional to Ca2+ entry

Transverse tubules connect plasma membrane of muscle cell to SR

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Ca2+ release during Excitation-Contraction coupling Action potential on motor endplate travels down T tubules
Ryanodyne R Ca-release ch.

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 Voltage -gated Ca2+ channels open, Ca2+ flows out SR into cytoplasm Ca2+ channels close when action potential ends. Active transport pumps continually return Ca2+ to SR

Ca ATPase (SERCA)

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Excitation-Contraction Coupling
Depolarization of motor end plate (excitation) is coupled to muscular contraction
Nerve impulse travels along sarcolemma and down T-tubules to cause a release of Ca2+ from SR Ca2+ binds to troponin and causes position change in tropomyosin, exposing active sites on actin Permits strong binding state between actin and myosin and contraction occurs ATP is hydrolyzed and energy goes to myosin head which releases from actin
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Summary: Excitation-Contraction Coupling

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IV Factors that Affect the Efficiency of Muscle Contraction

Tension and Load

The force exerted on an object by a contracting muscle is known as tension. The force exerted on the muscle by an object (usually its weight) is termed load. According to the time of effect exerted by the loads on the muscle contraction the load was divided into two forms, preload and afterload.

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Preload
Preload is a load on the muscle before muscle contraction.
Determines the initial length of the muscle before contraction.

Initial length is the length of the muscle fiber before its contraction.
It is positively proportional to the preload.
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The Effect of Sarcomere Length on Tension

The Length Tension Curve Concept of optimal length

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Types of Contractions I
Twitch: a brief mechanical contraction of a single fiber produced by a single action potential at low frequency stimulation is known as single twitch. Tetanus: It means a summation of twitches that occurs at high frequency stimulation
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Effects of Repeated Stimulations

Figure78 10.15

1/sec

5/sec

10/sec

50/sec

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Afterload
Afterload is a load on the muscle after the beginning of muscle contraction.
The reverse force that oppose the contractile force caused by muscle contraction.

The afterload does not change the initial length of the muscle,
But it can prevent muscle from shortening because a part of force developed by contraction is used to overcome the afterload.
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Types of Contractions (II)

Afterload on muscle is resistance Isometric
Length of muscle remains constant. Peak tension produced. Does not involve movement

Isotonic
Length of muscle changes. Tension fairly constant. Involves movement at joints

Resistance and speed of contraction inversely related

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Isotonic and Isometric Contractions

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Resistance and Speed of Contraction

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Muscle Power
Maximal power occurs where the product of force (P) and velocity (V) is greatest (P=FV)

Max Power= 4.5units

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