Smooth Muscle

Physiology
Jutta A. Ward, PhD
Reading: Blaustein et al.
Cellular Physiology and Neurophysiology
Ch. 15 241-246
Ch. 16 260-266
 Smooth Muscle Physiology Objectives
• Describe the differences in actin-myosin regulation of, respectively,
smooth, cardiac and skeletal muscle and indicate the structural
similarities in their respective contractile units.
• Explain why smooth muscles can develop and maintain force with a
much lower rate of ATP hydrolysis than skeletal muscle.
• Distinguish between muscle relaxation from the contracted state
and the phenomenon of stress relaxation and give examples of each
process.
• Diagram the intracellular pathways that control contraction and
relaxation in smooth muscle. Distinguish between
electromechanical coupling and pharmacomechanical coupling.
• Describe the distinguishing characteristics of multi-unit and unitary
smooth muscles.
 Lecture Outline
• Smooth muscle structure
• Smooth muscle innervation
• Smooth muscle action potentials
• Calcium
– Source of calcium
– Function of calcium in force of contraction and latch
mechanism
• Mechanisms of excitation
– Hormones
– Stretch
– Local factors
• Force-velocity relationships
 Smooth muscle structure
• Non-striated
• Spindle shaped, still have contractile
proteins actin and myosin, but no
sarcomeres so no striations
• Small (2-5 mm x 50-300 mm)
• 1 centrally placed nucleus
• Thin filaments arranged in in
bundles and attached to dense
bodies (a-actin) on either the
sarcolemma or cytoskeleton
• Thick filaments inter-digitate among
thin filaments
G & H Fig 8-2
 Smooth muscle structure
• Sarcoplasmic reticulum: Poorly
developed and sparse, no t-tubule or
terminal cisternae so
• Both extracellular and intracellular
Ca2+ initiate contraction.
• Regulated by autonomic nervous
system and hormones

G & H Fig 8-5

 Smooth muscle contractile properties
• Slow cycling of myosin cross-bridges
– Much slower: cycling occurs 1/10 to 1/300 the rate of skeletal
muscle
– ↑ time of cross-bridge attachment.
• Low energy requirement
– Due to fewer cycles less ATP is split
• Slow onset and relaxation of contraction
– Does not reach full contraction until 0.5 sec
– May not relax for 1-3 sec or up to 30 sec
– Again, slow cross bridge cycling coupled with delayed
response to Ca2+ ions
 Smooth muscle contractile properties
• Force of contraction
– Force may be as high as 4-6 kg/cm2
– Skeletal muscle max force ~3-4 kg/cm2
• Stress relaxation
– Allows hollow organs to expand and contract as needed
– Muscle contracts or relaxes at an appropriate level to maintain
pressure with in the organ
• ie. Filling and emptying of bladder
• “latch” mechanism (a tonic contraction)
– Large and sustained contraction with low energy expenditure
 Lecture Outline
• Smooth muscle structure
• Smooth muscle innervation
• Smooth muscle action potentials
• Calcium
– Source of calcium
– Function of calcium in force of contraction and latch mechanism
• Mechanisms of excitation
– Hormones
– Stretch
– Local factors
• Force-velocity relationships
 Smooth muscle: multi-unit

• Each fiber can contract

independently and is usually
innervated by a single nerve ending
• no electrical coupling makes neural
G & H Fig 8-1

integration important
• Fibers insulated from each other by
a covering of collagen and
glycoprotein fibrillae
• Seldom exhibit spontaneous
contraction
• Examples: ciliary muscle of eye,
B &B iris, piloerector muscles
Fig 9-2
 Smooth muscle: unitary
• Aggregates (bundles or sheets) of hundreds or
thousands of fibers act as a single unit
• Membranes are adherent to one another at
numerous points so that the force generated in
one fiber can be transmitted to the next.
• Fibers are connected by gap junctions which
allow ion flow from one fiber to another such
that electrical activity is propagated throughout
the tissue. Therefore action potentials and
contractions are simultaneous
• Examples: walls of viscera, gut, bile ducts,
ureters, uterus and many blood vessels
 Innervation of smooth muscle
• Most smooth muscles probably arranged in the single-unit
fashion.
• Contraction (or inhibition of contraction) of smooth muscles can be
initiated by
– (1) intrinsic activity of pacemaker cells
– (2) neurally released transmitters
– (3) circulating or locally generated hormones or signaling molecules. The
combination of a neurotransmitter, hormone, or drug with specific
receptors alters contraction by increasing or decreasing intracellular Ca2+.
Single-unit:
most common

Multi-unit
Berne et al, fig 13-7
 Lecture Outline
• Smooth muscle structure
• Smooth muscle innervation
• Smooth muscle action potentials
• Calcium
– Source of calcium
– Function of calcium in force of contraction and latch
mechanism
• Mechanisms of excitation
– Hormones
– Stretch
– Local factors
• Force-velocity relationships
 Membrane and action potentials

• Typical smooth muscle resting potential -50 to -60 mV

• Ca2+ plays a role in the generation of the AP
– Smooth muscle has many more voltage-gated (L-type) Ca2+
channels than skeletal muscle.
– Less participation of Na+ in action potential
– Ca 2+ channels open slower and stay open longer allowing
some cells to have plateaus in their AP’s
 Types of AP seen in smooth muscle
Seen in unitary (visceral) smooth muscle
• Slower upstroke and longer duration than skeletal muscle
• Upstroke due to Ca2+ channels. Inward Ca2+ current depolarizes cell
causing more voltage gated Ca2+ channels to open. However, Ca2+
channels are slower than voltage gated Na+ channels so everything occurs
more slowly.
• Spike potential
– Elicited via electrical, transmitter, hormonal or stretch stimulus
– Last up to 50msec
• Plateau potential
– Can last from 100 – 1000 msec
• Slow wave potentials
Some smooth muscles contract without an AP
• Oscillations in Vm may be sufficient for tonic
contraction even without an AP.
• Hormones and neurotransmitters may cause
contraction with no change in Vm. Instead
second messengers are produced that
increase cytosolic Ca2+ release from the SR.
• It’s all about Ca2+ availability!
 Lecture Outline
• Smooth muscle structure
• Smooth muscle innervation
• Smooth muscle action potentials
• Calcium
– Source of calcium
– Function of calcium in force of contraction and latch mechanism
• Mechanisms of excitation
– Hormones
– Stretch
– Local factors
• Force-velocity relationships
 Sources of calcium
1. voltage-gated L-type Ca2+ channel
2. SR release without depolarization
3. Ca2+ entry via non voltage-gated channel
 Calcium release from SR

• Skeletal muscle
Fig. 15-7

• Cardiac and
Smooth muscle

Fig. 15-11
Cellular Physiology and Neurophysiology, Third Edition
 Many roles for Ca2+ in smooth muscle
Caldesmon, together with tropomyosin, regulate the access of myosin to
actin. In relaxed smooth muscle the binding site on myosin is blocked by
the caldesmon-tropomyosin complex.

Calponin is a smooth muscle-specific, actin-, tropomyosin- and calmodulin-binding

protein involved in regulation or modulation of contraction
 Role of Ca2+ to activate thick filaments

B & B Fig 9-17

• Ca2+ required to increase myosin ATPase activity

• Four Ca2+ bind calmodulin (CM) which is related to Tn-C)
• CaCM complex activates myosin light chain kinase (MLCK)
• The kinase phosphorylates the regulatory light chain on the myosin
head.
• The phosphorylation event increases ATPase activity of myosin
• Contraction requires a phosphorylation of the myosin regulatory chain
 Termination of contraction
• MLC Kinase phosphorylates the MLC allowing
contraction.
• Even with the removal of calcium, MLCK may
become inactive, but as long as the MLC is
phosphorylated, contraction can continue.
• Relaxation requires de-phosphorylation of the
MLC by MLC phosphatase.
 Lecture Outline
• Smooth muscle structure
• Smooth muscle innervation
• Smooth muscle action potentials
• Calcium
– Source of calcium
– Function of calcium in force of contraction and latch mechanism
• Mechanisms of excitation
– Hormones
– Stretch
– Local factors
• Force-velocity relationships
 Regulation of smooth muscle actin-myosin
interactions by Ca2+stimulated phosphorylation.
Berne et al.Fig 13-8

• In the relaxed state cross-bridges are

present as a high-energy myosin-ADP-Pi
complex in the presence of ATP.
Attachment to actin depends on
phosphorylation of the cross-bridge by a
Ca2+-calmodulin-dependent myosin light
chain kinase (MLCK). Phosphorylated
cross-bridges (color) cycle until they are
dephosphorylated by myosin
phosphatase. ATP binding and
myosin detachment
• Note that cross-bridge phosphorylation
at a specific site on a MLC requires ATP
in addition to that used in each cyclic
interaction with actin.
CM: Calmodulin.
• Covalent regulation allows eight cross-bridge states in smooth
muscle. Phosphorylation by MLCK (vertical-colored arrows) is
obligatory for cross-bridge attachment. Phosphorylated cross-
bridges (color) cycle comparatively rapidly. Dephosphorylation
of a cross-bridge during a cycle by a constitutively active
myosin phosphatase (vertical black arrows) slows cycling
rates and produces the latch state.

Calcium regulates cross-

bridge cycling by
determining
phosphorylation rates.

Note that ATP is required

for both regulation
(vertical arrows) and
cycling (curved arrows).

Berne, et al Fig 13-10

De-phosphorylation slows activity of
myosin ATPase
 Events in cross-bridge activation and
contraction in smooth muscle.
• Phasic contraction: A brief period of
stimulation is associated with Ca 2+
mobilization followed by cross-bridge
phosphorylation and cycling to
produce a brief phasic, twitch like
contraction.

• Tonic Contraction: In a sustained

tonic contraction produced by
prolonged stimulation, the Ca2+ and
phosphorylation levels typically fall
from an initial peak. Force is
maintained during tonic contractions
at a reduced Ca2+ concentration (and
hence a low level of myosin light
chain phosphorylation), with lower
cross-bridge cycling rates manifested
by lower shortening velocities and
ATP consumption.

Berne et al.Fig 13-9

 Latch
mechanism

Dephosphorylation of the
myosin light chain by
myosin light chain
phosphatase (MLCP)
helps maintains tonic
contraction with reduced
cross bridge cycling and
therefore extremely
low energy expenditure
Non-electrical stimulation of smooth muscle via
G-protein coupled ER release via IP3 pathway
 1. Ca2+ enters the cytoplasm through
Cav channels located in caveolae
STIC (spontaneous transient inward currents)
2. Agonists, acting through a G- STOC (spontaneous transient outward currents)
protein–coupled receptor can lead to STICs induce depolarization, whereas STOCs hyperpolarize
the formation of IP3 the membrane potential

4
3
4. Depletion of SR Ca2+ stores causes
aggregation of STIM1 monomers in the SR 3. Ca2+ entering via Cav channels
membrane, thereby activating Orai channels in activates RYR, which release more Ca2+
the plasma membrane, allowing Ca2+ to enter via CICR. IP3 formed by PLC activates IP3R,
the cytosol for uptake into the SR by SERCA which also release Ca2+. Both pathways
contribute to Ca2+ sparks
 The transport systems that participate
in the regulation of [Ca 2+ ] i in smooth
muscle cells
• Ca 2+ can enter the cells through DHPRs,
receptor operated calcium channels (ROCs),
stretch activated calcium channels (SOCs),
and the sodium-calcium exchanger (NCX).
• Cytosolic Ca 2+ is extruded from the cells by
plasma membrane calcium ATPase (PMCA)
and the NCX and pumped into the SR by
sarcoplasmic reticulum calcium ATPase
(SERCA).
• Ca 2+ is released from the SR through
channels gated by IP 3 (i.e., IP 3 Rs), and
through RyRs that are opened by elevated
[Ca 2+ ] i and that mediate calcium induced
calcium release (CICR)
 Mechanisms determining myoplasmic
[Ca2+] in smooth muscle.Berne et al. Fig 13-11
• Ca2+ release from the SR is a
rapid initial event in activation,
whereas both the SR and the
sarcolemma participate in the
subsequent stimulus-dependent
regulation of myoplasmic [Ca2+].
• The sarcolemma integrates many
simultaneous excitatory and
inhibitory inputs to govern the
cellular response. Higher-order
regulatory mechanisms can alter
the activity of various pumps,
exchangers, or enzymes (the
asterisks designate well-
established instances).

G, Guanine nucleotide binding proteins; PL C, phospholipase C; PIP2, phosphatidylinositol bisphosphate;

IP3, inositol 1,4,5-trisphosphate; CM, calmodulin; MLCK, myosin light chain kinase
 Both Ach and Norepi are released at the
smooth neuromuscular junctions
Somatic neuron: the pathway between the
CNS and effector cell is monosynaptic. The
neuron releases (ACh), which binds a nicotinic Major neuro-transmitters of
receptor on the skeletal muscle cell). the autonomic nervous system.

ANS: both parasympathetic and sympathetic

preganglionic neurons release ACh, which
binds a nicotinic receptor on the postsynaptic
membrane of the postganglionic neuron.

Parasympathetic postganglionic neurons:

the neurotransmitter is ACh, but the
postsynaptic receptor is a muscarinic
receptor, which is a metabotropic (i.e., G-
protein-linked)

Sympathetic postganglionic neurons, the

neurotransmitter is norepinephrine. The
postsynaptic receptor is a metabotropic (i.e.,
G-protein-linked) adrenergic receptor of one
of two major subtypes (α and β).

B & B Fig 15-8

 Lecture Outline
• Smooth muscle structure
• Smooth muscle innervation
• Smooth muscle action potentials
• Calcium
– Source of calcium
– Function of calcium in force of contraction and latch mechanism
• Mechanisms of excitation
– Hormones
– Stretch
– Local factors
• Force-velocity relationships
Hormonal effects excite and inhibit
• Diversity of effects: norepi inhibits smooth muscle
contractions in the gut but stimulates contraction in vascular
smooth muscle.
• Excitation: hormone binds ligand gated Na+ or Ca2+ channels
and depolarize the membrane.
• Inhibition:
– ligand gated Na+ or Ca2+ channel is closed
– the normally closed K+ channel is opened (Ca2+ activated K+
channels).
– In both cases membrane potential becomes more negative
inside making it more difficult to initiate contraction
– Hormone may bind to membrane and indirectly inhibit via
cAMP and cGMP second messenger systems.
• Indirectly activates Ca2+ pump in cell membrane and SR
• Net effect is decreased Ca2+ in sarcoplasm
Multiple mechanisms of control in smooth
muscle
 What in the world is a cell to do?
Its all so confusing…
Selected action
Receptor Agonist potency order Mechanism
of agonist
Gq: phospholipase C (PLC)
α1: Norepinephine > Smooth muscle
activated, increased IP3 and
Epinephine contraction
increased intracellular [Ca2+]

Smooth muscle
α2: constriction and Gi : adenylate cyclase
Epi ≥ norepi
neurotransmitter inactivated, decrease in cAMP
inhibition

epinephrine = Heart muscle Gs: adenylate cyclase activated,

β1
norepinephrine contraction increased cAMP

epinephrine >> Smooth muscle Gs: adenylate cyclase activated,

β2
norepinephrine relaxation increased cAMP
Catecholamine modulation of vascular smooth
muscle contraction
• Vascular smooth muscle has β2-
adrenoceptors that have a high binding
affinity for circulating epinephrine and a
relatively lower affinity to norepinephrine
released by sympathetic adrenergic nerves

• These receptors are coupled to a Gs-protein

which stimulates the formation of cAMP. An
increase in cAMP leads to smooth muscle
relaxation. The reason for this is that cAMP
inhibits myosin light chain kinase that is
responsible for phosphorylating smooth muscle
myosin. Therefore, increases in intracellular
cAMP caused by β2-agonists inhibit myosin light
chain kinase thereby producing less contractile
force (i.e., promoting relaxation).
 B2, A2,

From: cvphysiology.com
Excitation of unitary muscle due to stretch

• Stretching of visceral smooth muscle may

generate spontaneous action potentials via
– Stretch-activated cation-permeable channels
– Decrease in overall negativity of the membrane
caused by the stretch itself
• Allows gut wall to contract automatically
and rhythmically when stretched
excessively (ie peristaltic waves).
 Contraction due to local tissue factors

• In the vasculature, the smallest arterioles have

smooth muscle but little innervation
• Instead blood flow control is via local tissue
factors that cause vasodilation
– Lack of O2
– Excess CO2
– Increased H+
– Other factors include increased adenosine, lactic
acid, K+ ions and body temp
 Lecture Outline
• Smooth muscle structure
• Smooth muscle innervation
• Smooth muscle action potentials
• Calcium
– Source of calcium
– Function of calcium in force of contraction and latch mechanism
• Mechanisms of excitation
– Hormones
– Stretch
– Local factors
• Force-velocity relationships
 Force-velocity curves for fast and slow skeletal
muscle cells and smooth muscle.
• B: Smooth muscles have variable
force-velocity relationships that are
determined by the level of Ca2+-
stimulated cross-bridge
phosphorylation.
• C, Maximal shortening velocities
with no load (intercepts on the
ordinate in B) are directly
dependent on cross-bridge
phosphorylation by MLCK.
• D, Active force/stress (abscissa
intercepts in B) rises rapidly with
phosphorylation, and near maximal
stress may be generated with only
20% to 30% of the cross-bridges in
the phosphorylated state.

Berne et al. Fig 13-13