Chapter 7

Excitation of Skeletal Muscle

Neuromuscular Transmission
Excitation-Contraction Coupling
§7.1 Neuromuscular
Transmission
Experiment

How is information transmitted from

nerve to the muscle?
 Structure of the
Neurotransmission Junction
 The neuromuscular junction (NMJ) is a
highly-specialized neurotransmission
system between motor nerve and muscle.

 Its role is reliable one to one translation of

motoneurone action potentials into muscle
action potentials.
The essential structural features of the NMJ

1. Prejunctional
membrane
2. Junctional cleft
(50 nm)
3. Postjunctional
membrane (end
membrane or
motor end plate)
4. Synaptic vesicles
5. AChR
 Discovery of Neuromuscular
Transmission
 The experiment demonstrates that there is a
delay between the arrival of the AP at the
nerve terminal and the development of the
muscle AP. F
 Muscle event can be selectively suppressed
by bathing the nerve and muscle preparation
with a solution containing a drug called
curare .
 Surface Recording at a NMJ

A. An electrical signal
which produces a
artifact.
B. An action potential
detected at the
electrode.
C. The massive muscle
fiber action potential
(0.5 to 0.75ms later)
Obviously, there is a chemical
link which can be affected by
curare between motoneurone
AP and the muscle AP……
Now , we know that is
acetylcholine (ACh)
Acetylcholine synthesis,
storage and release
ACh is concentrated and
stored in synaptic vesicles.
4
Each vesicle stores ~ 10
molecules.

These loaded vesicles are

released as the basic
quanta, or packets, of the
transmission process.
Acetylcholine receptor
ACh is broken down by AChE
 Events occurred at the
Neuromuscular Junction
An AP depolarized axon terminal
AP
↓ The depolarization opens voltage-
gated Ca channel and Ca enters
the cell from ECF

Ca entry triggers exocytosis of

synaptic vesicle contents

↑Ca ACh diffuses across the

synaptic cleft and binds with
the α subunits of nicotinic
receptors on the motor end
plate
 The binding causes a conformational
change in α subunits of receptor

AP The center core of the channel opens

↓ and the permeability of the motor end
plate to both Na & K increases.

Both Na & K flow diffuse downhill,

resulting a depolarization of the motor
end plate, called the end plate potential
(EPP, around 0mV).
↑ Ca
With a effect of EPP, adjacent
area of the muscle fiber is
depolarized to threshold by the
spread of local current, and they
fire action potentials.
 Miniature endplate potential
At rest, single packets are
released at random intervals
at a rate of about 1per
second. This spontaneous
release causes small, 0.4 mV,
deplolarizations of the motor
endplate called miniature
end plate potentials (MEPP).
Some Questions Concerning
Neuromuscular Transmission
 NMJ is a site where energy was converted:
Electrical→chemical→electrical
 EEPs are elicited at the NMJ when ACh
activates the nicotinic AChR.
 These local potentials are large enough to
initiate an action potential and contraction
in the skeletal muscle (~50 mV in amplitude).
 Receptorsdirectly gate ion channels. The
ion channel is part of the receptor.
 Cholinergic nicotinic (N2 type) receptor
is classified as a nonselective cation
channel ： permeate to K ＋ efflux
( hyperpolarizing) and Na ＋ influx
(depolarizing).
 Characteristics of
transmission in NMJ
 Propagated from initial part along axon all
the way down to its end
 Transmission with 0.5 to 1 ms delay
 One action potential arising from axon
ensured one action potential in sarcolemma
membrane (1:1transmission)
 Affected by some factors
 Factors That disturb
transmission in NMJ
 The factors that decrease ACh release :
decrease[Ca2+]O or increase [Mg2+]O;
Botulism toxin.
 Decrease number of cholinergic receptors ：

myasthenia gravis.
 AChR is blocked by some toxin (curare, snake toxin)
 Decreased or damaged AChE enzymatic activity:

Organophosphate and Carbamate Poisoning

 Myasthenia Gravis

The muscular weakness and fatigability

associated with myasthenia gravis are
caused by an autoimmune attack on the
AChR at the neuromuscular junction.
Antibodies have been shown to decrease
the usefulness of acetylcholine receptors
through accelerated endocytosis and
blockade of the receptor.
 Distinguish an EPP and an AP
in the Skeletal Muscle
• AP propagate in a  EPPs are non-regenerative
regenerative manner, in signals generated in the
an “all or none” fashion vicinity of the neuromuscular
along the skeletal muscle junction by the opening of
membrane ultimately ligand-gated channels.
stimulating contraction EPPs depolarize the skeletal
• AP generated is due to muscle membrane to threshold
opening Na+ channel on to generate an AP.
the muscle membrane.
 §7.2 Excitation–
Contraction Coupling

Concept: The mechanism by which an

AP produced in the sarcolemma at the
NMJ triggers contraction of a muscle
cell is called E-C coupling.
 How does the AP cause 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?
 Terminal cisterna

Triad T-tubule

Terminal cisterna
Longitudinal SR (LSR) Junctinal SR (JSR)
Containing Ca2+ pump Containing RyR
 Transverse tubules

 To conduct the AP from the surface,

deep into the muscle fibre -
– the t-tubules are continuous with
the sarcolemma and are filled with
extracellular fluid i.e. Na+ + Ca2+
rich
 To signal the passage of the AP to
the SR in some way???
 Sarcoplasmic reticulum

1. To store very high concentration of Ca2+

- bound to calsequestrin
2. To release Ca2+ in response to an AP in
the t-tubules
3. To remove (re-sequester) the Ca2+ in
order to stop contraction
 Where intracellular Ca2+
comes from during contraction?

 Influx
through
L-type Ca
channel

 Release
from SR
via RyR
(ryanodine receptor)
 E-C Coupling in
The AP:
Skeletal muscle
moves down the t-tubule T-tubule
sarcolemmaout
the voltage change is sensed
in
by the DHP receptor sarcoplasmic
reticulum
–DHP receptor is essentially a
voltage-gated Ca channel

is communicated to the

ryanodine receptor which opens
Ca then:
voltage sensor
–floods out of the SR (DHP receptor) junctional foot
(ryanodine receptor)
–activates contraction
 The Answers
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
 Ryanodine Receptors and
Ca Release
2+

•complexes
Ryanodine receptor (RyR) is a protein
that mediate the release of
calcium from the sarcoplasmic reticulum
(SR,) in both skeletal and cardiac muscle
cells by forming tetrametric complexes.

•CaThese
2+
complexes each then act as a
channel.
 Three Isoforms of the RyR

• RyR1 is specifically
expressed in skeletal
muscles

• RyR2 in cardiac muscles.

• RyR3 is another isoform

found in non- muscle cells
such as neuronal cells.
 Where the Ca2+ goes while
muscle relaxation?
 Reuptake into SR by SR Ca-ATPase and
stored in cisternae by binding to a protein,
calsequestrin
 Remove out of cell by Na+/Ca2+ exchanger
 Remove out of cell by sarcolemma Ca2+
pump
 Comparison
Skeletal muscle Cardiac muscle
[Ca2+ ] 100% from SR 80% to 90% from SR,
↑ 10% to 20% influx via L-
type Ca2+ channel
[Ca2+ ] ↓ By SR pump By SR pump, Na+/Ca2+
exchanger, pump of the
cell membrane
RyR RyR1; RyR2
Ca2+ channel : RyR1 = 1:1 Ca2+ channel : RyR2 =1:7 to 10

CICR none yes

 Chapter 6

Contraction of
Skeletal Muscle
 Outline
 Physiological anatomy of skeletal muscle

 Molecular mechanism of muscle contraction

 Effect of actin and myocin filament overlap

on tension developed by contacting muscle
Structure of muscle
The parts of a muscle

Skeletal muscle
Muscle fibers

Nuclei
Cytosole
Myofibrils

Thick filament
Filament
Thin filament
Sarcomere: the basic unit of contraction
Structure of a sarcomere

1. Actin and myosin filaments(A-band,)

2. Myosin filaments only (H-zone)
3. Actin filaments only (I-band)
4. M line
5. Z line
 Structure of Myofilament
Head consists of 4 light,
two heavy chain
Thick filament: Myosin
Myofilament

F Tail consist of two

heavy chains C

Actin has binding sites of

myosin
Thin filament Tropomyosin
F R
Trponin has affinities for actin,
tropomyosin and Ca2+
Actin binding site
ATP binding site
 Actin has binding sites of myosin

Tropomyosin
Thin filament
Trponin has affinities for actin,
tropomyosin and Ca2+
 Mechanism of contraction
Sliding filament theory
When muscle shorten, the
filaments of action and
myosin which make up a
sarcomere do not shorten;
rather they slide past each
other like the fingers of two
hands interdigitating.
 The Molecular Basis of
Contraction
The Crossbridge Cycle
The crossbridge cycle refer to a
process of the repeated binding,
shortening, releasing and re-
binding of crossbridge.
The energetic of filament sliding
1. During crossbridge cycle, myosin converts the
chemical energy of ATP into the mechanical
energy of filament sliding.
2. Each cycle of mechanical activity of the
myosin crossbridge takes about 50 ms and is
accompanied by a cycle of ATPase activity.
3. During a contraction, each myosin head
undergoes a conformational change that moves
the thin filament 5 to 15 nm during a period as
shorter as 50ms.
The crossbridge cycle
Six Steps of Crossbridge Cycle
1. The influx of calcium, triggering the
exposure of binding sites on actin
2. The binding of myosin to actin
3. The power stroke…
4. The binding of ATP
5. The hydrolysis of ATP...
6. The transport of calcium ions
back into the sarcoplasmic
reticulum
 §3.4 Analysis of the force
generated by contraction
当肌肉克服某一外力而缩短或肌肉缩短
牵动负荷时，肌肉作了一定量的机械功
，其数值可以下式表示：

功 (W) ＝克服的阻力（或负荷）　肌肉缩短的
长度　
 Types of contraction of
skeletal muscle

Resting tension ( 静息张力）

Isometric contraction ( 等长收缩 )
Isotonic contraction ( 等张收缩）
 Tendon ( 肌腱）

Non-contractile and connective

tissue in muscle itself
 Isometric Contraction
Increase in muscle tension without a
change in muscle length ( 张力增加
，没有长度改变的收缩 ).

Since L=0 so, W=0

wall
Energy→tension
 Isotonic Contraction
Contraction in which the tension remains
constant as the muscle shortens or lengthens
( 张力不变，只有长度改变）

W=L× load L

load
Isotonic Isometric
 Preload and Afterload
Preload( 前负荷 ): load establishes the
initial muscle length. Load is carried by
muscle at rest and during contraction.
　　（前负荷使肌肉收缩时在一定初长基础上开始）
Afterload ( 后负荷 ): load, or a portion
of it, is supported until the muscle
develops enough force to lift it.
　（后负荷不改变肌肉的初长，但阻碍肌肉收缩时长度的改变）
  Effects of preload or
initial length on contraction
of the muscle 　
 Initial length & preload
Move up or down to change preload (initial length)

F2
S
F1

F3

Transducer
1. Passive tension( 被动张力 ) is the
tension developed by simply
stretching a muscle to different
lengths ( think of a rubber band).

2. Passive tension is determined by

preload and the elasticity of the
muscle itself.
Total tension( 总张力 ) is the tension
developed when a muscle is stimulated
to contract at different preload.
It is the sum of the active tension
developed by contractile elements of
the sarcomeres and the passive tension
caused by stretching the muscle.
Active tension ( 主动张力 )is determined by
subtracting the passive tension from the total
tension (total T-passive T).
It represents the active force developed when
the muscle contracts, i.e., the relation between
preload and tension generated by contraction.
 At the beginning, active tension developed is
proportional to the increase in preload but,
when the muscle is stretched to longer length,
active tension is reduced. B
Optimal preload or initial length
（最适前负荷或最适初长度）
前负荷增加到某一限度，使肌肉
收缩张力开始下降的临界值

Why ？
The active tension is maximal when there is maximal overlap
of thick and thin filaments and maximal cross-bridge

2 to 2.2µ m in
vivo
Latency & the distance
shortened

Increasing preload
does not affect
latent period and
slightly reduces the
distance shortened
 Effects of afterload on
the contraction of
muscle:
Force-Velocity
relationship
 Experiment 在有后负荷的情况下，肌肉
能否缩短、其缩短速度，取
决于肌肉收缩产生的张力与
负荷的力量对比　

9
Afterload 5
3
Preload

Tension transducer
Latency & the distance
shortened
Increasing afterload

increase the latent

period and reduces

the distance shortened

 Because:
1. The speed of muscle shortening depends
on the speed of cross-bridge cycling.

2. The force developed depends on the

number of cross-bridge formed.

3. As the afterload on the muscle increases,

the velocity will be decreased because
cross-bridge can cycle less rapidly
against the higher resistance.
 Force–Velocity Curve
2. 当 P<P0, 肌肉收
缩时既产生张力
又出现缩短；
3. 一旦产生的张力
＝负荷，肌肉得
P 以缩短，移动负
荷。此时张力不
P0 再增加即为等张
收缩；
4. 肌肉承受的后负
荷越小，等张收
1. P0 表示后负荷增加到某一数值 缩产生的张力越
，肌肉产生最大张力但不能缩短 小，而缩短速度
， Vmax=0; (isometric 却越快。
contraction)
负荷对肌肉收缩的影响

❧负荷越大，肌肉收缩
时
•最终产生的张力越大
•出现缩短的时间越晚
•缩短的程度越小
•缩短的速度越小
 Afterload & Power
POWER = FORCE x VELOCITY
physiological power = strength of muscle
contraction x velocity of muscle contraction
P=L× V
1. Muscle fiber can
shorten rapidly or
develop high forces
but not at the same
time
2. Peak power occurs
at approximately 1/3
maximum
shortening velocity.
Effects of contractility on
the contraction of muscle
Contractility is determined by the level of intracellular
Ca2+ , the activity of myosin and ATPase, etc. In vivo, the
contractility of the muscle is regulated by hormones,
drugs, and some other humoral substances.
The difference of muscle fiber
 Summation of Contraction
Single twitch & tetanus
单收缩和强直收缩
Intracellular Ca

tension
Question
1. When high rates of stimulation produce
complete fusion of contractions, tetanus, is
the action potentials that generated
contraction able to be fused? （当高频刺激使肌
肉产生强直收缩时，触发收缩的 AP 会发生融合吗？）
2. When a stimulation apply to a preparation of
nerve-muscle, what will be happen?
Question

2. What is the intracellular Ca store called?

3. What function does the t-tubular system serve?

4. What determines force of contraction for a
piece of muscle.
 APs are propagated to the T tubules
by the spread of local current
↓
Depolarization causes a conformational change
in its DHP receptor and then opens the Ca2+
release channels (RYR) on the near by SR
↓ ↓
Ca2+ is released from SR SR Ca2+ ATPase is activated
↓ ↓
Ca2+ binds to TnC, Tropomyosin Ca2+ is re-accumulated
is moved out of the way in SR, resulting [Ca]i ↓
↓ ↓
Contraction Relaxation
 L-type Calcium Channels

 L-type calcium channel is also called

dihydropyridine (DHP) receptor.
 It is a kind of voltage-gated channel
contained 4 or 5 distinct subunits ( α 1, α
2, β , δ , or γ ).
 Ca plays the important role of cross-talk
between T tubule and RyR in the process of
muscle contraction.