Contractile Mechanisms 1 - Basics of

Muscle Physiology / Basics of Modeling

Muscle Function


J. Jeremy Rice
IBM T.J. Watson Research Center, P.O. Box 218,
Yorktown Heights, NY 10598
johnrice@us.ibm.com
914-945-3728


Goals
Philosophy and motivation
Basic machinery
Contractile proteins
Sarcomere structure
Length-tension relationship
Classical models

Philosophy and motivation
Multiple views of muscle
Schematics
Primary data sources from published literature
Protein structures (dont need to know for test)
Histology (dont need to know for test)
Engineering descriptions
Mix pf both historic and current data
Be critical thinkers about models
Conceptual versus quantitative models
Animation versus simulation
Successes and limitations
Proud of how smart you will be after lectures



Basic machinery
(Big picture)
Cyclical interactions between actin
and myosin produce force/motion
+ -
1) Attach 2) Rotate head
- stretch spring
3) Detach and
rotate back
4) Repeat
Thin filament - actin
Thick filament - myosin
Cyclical actin and myosin interactions converts
energy in ATP to force/motion
From the website of the Michael
Geeves' Laboratory:
http://www.kent.ac.uk/bio/geeve
s/Research/home.htm
Actin and myosin are arranged in
a regular lattice of units called
sarcomeres
+
-
+
-
Sarcomere
becomes shorter
from actin-myosin
interactions
More detailed look at
contractile proteins

Alternative view of myosin
structure
Tail extends here to thick filament
Alternative view of cross-
bridge cycle
Position of myosin heads on
thick filament
Pairs of heads emanate 180
degree apart in radial direction at
each step
Radial direction of heads rotate
~60 degrees in radial direction at
next step in axial direction
(distance = ~14.3 nm)
a "pseudo-repeat" happens on
the 3rd step as heads will emanate
in same radial direction (distance =
~43 nm)
a
x
i
a
l

d
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radial direction
Actin reconstruction from Schutt et al., 1994
for beta actin-profilin complex, see
www.med.ic.ac.uk/divisions/32/cmjulie.htm

Scheme for the structure of actin. (From
Holmes and Kabsch, 1991).

Actin is a polymer chain
+
-
Thin filament is a two-stranded helix of
actin monomers
From http://www.kent.ac.uk/bio/geeves/Research/home.htm
"Pseudo-repeat" = 13 units
5.54 nm
2.77 nm
"Psuedo-repeat" 37 nm
True repeat = 26 units
Actin can be "decorated"
with rigor myosin heads
3D maps illustrate actin-myosin
interactions from cryo-EM and image
analysis. F-actin is red, tropomyosin is
blue, the myosin motor domain is yellow,
the essential light chain is blue-green, and
the regulatory light chain is dark red.
From Ron Milligan Lab
http://www.scripps.edu/milligan/

An Animated Model for Muscle Myosin-Based Motility
After completing the stroke, ADP dissociates and ATP binds to the empty active site,
which causes the lever arm to recocks back to its prestroke state (transition from red
to yellow). The surface features of the myosin head and the actin filament were
rendered from X-ray crystal structures by Graham Johnson (fiVth media:
www.fiVth.com) using the programs MolView, Strata Studio Pro and Cinema 4D.
Transitions between myosin crystal structure states were performed by computer-
coordinated extrapolations between the known prestroke and poststroke positions as
taken from PDB (Protein Database).

Animation by Ron Vale, Ron Milligan, and Graham Johnson
The Way Things Move: Looking Under the Hood of Molecular Motor
Proteins Ronald D. Vale and Ronald A. Milligan Science Apr 7 2000: 88-95.

Motility assay

Motility assay
Sarcomere structure


Banded Structure of Sarcomere
M-band
H zone
M-band
Band definition in skeletal muscle
In each sarcomere, the broad band which appears dark in standard histological
procedures is called the A band. This band indicates the location of thick filaments
(myosin); it is darkest where thick and thin filaments overlap (called the A I band).
The broad light band between the dark bands is the I band. The I band indicates the
location where thin filaments (actin) extend beyond the thick filaments.
A distinct dark line running down the middle of the I-band is the Z line, where thin filaments
are attached end to end. Where thick filaments are attached end-to-end in the center of the
A-band is the M line.
When muscle is stretched, an H band appears along the middle of the A-band, between
the free ends of the thin filaments.
EM Microscopy - Evidence of Crossbridges
Huxley & Hanson, 1954
A bands are composed of actins and myosin
I bands are composed of actin
A bands do not change length over most of the sarcomere length range
Sliding filament theory is proposed
H.E. Huxley coins the phrase cross-bridges
Component proteins in skeletal
sarcomere
Tropomodulin (end cap)
M-band
Titin (think of most giant protein)
Span from M-line through thick filament to Z line
Giant protein made of repeats of immunoglobulin (Ig)-like domains
Act like a big spring as Ig-domain like to stick together (perhaps actin also)
Appears to interact with myosin in the thick filament

Granzier and Labeit, Circ. Res. 2004; 94: 284-295
Cardiac Z-disk
Modified from Pyle and Solaro, Circ. Res. 2004;94:296-305.
Z- disk
Titin sets slack length of sarcomere
Modified from Preetha et al., J Muscle Res Cell Motil. 2005;
26(6-8):307-17.
Below slack length
(B) Above slack length
(A) At slack length
(C) Full extension in cardiac muscle (~ 2.4 mm)
Components of sarcomere
Thick filaments
myosin
titin (provides template for thick filament, produces
restoring forces, sets a default sarcomere length)
Cardiac myosin binding protein-C (Slide Set III)
Thin filaments
actin, troponin, tropomyosin (Slide Set III)
nebulin (acts as ruler/template, skeletal only), nebulette
(interacts with z-disk, cardiac only), tropomodulin (end
cap that limits growth of thin filament)
Z disks (outside border of sarcomere, matrix of proteins)
Myosin Binding Protein C (modulates actin-myosin
interactions)
Other proteins for regulation (dont worry - not on test)
Force-length relations

Ramsey and Street, 1940 Frog skeletal muscle
Proposed structure-function
relationship model
Complete skeletal F-SL
relationship
Evidence for different F-SL relationship in
cardiac muscle vs. skeletal
pCa=4.6
Rat, isolated cardiac myocytes, from
Weiwad et al., 2000 note that
isolated cell with collagen removed
can be stretched past 2.3-2.4 mm
maximum length seen in trabeculae
Rat, 25C, skinned ventricular trabeculae,
from Wannenburg et al., 1997 Note that
force continues to rise in the range of 2.0
to 2.2 mm that is the plateau in the skeletal
F-SL response
pCa=5.3
pCa=5.6
F
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c
e

Complete cardiac F-SL
relationship
A. Assumed lengths of filaments
2.4 mm
2.3 mm
1.9 mm
1.6 mm
1.4 mm
1.6 mm
0.1mm
1.2 mm
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4
Sarcomere length (mM)
N
o
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m
a
l
i
z
e
d

f
o
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c
e

a
n
d

O
v
e
r
l
a
p

F
r
a
c
t
i
o
n

(
u
n
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l
e
s
s
)
From Rice et al., Biophysical Journal 2008
B. Assumed active force reflects the
single overlap fraction of thick filament
(mm)
F-SL relationship in cardiac
muscle
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4
Sarcomere length (mM)
N
o
r
m
a
l
i
z
e
d

f
o
r
c
e

a
n
d

O
v
e
r
l
a
p

F
r
a
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t
i
o
n

(
u
n
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e
s
s
)
Active Force
Passive Force
Note that the passive force includes the
contribution of collagen that gets very stiff
above 2.3-2.4 mm in trabeculae
(adapted from de Tombe & ter Keurs, 1990)
(mm)
Contribution of titin and collagen to passive myocardial tension of mouse
and dog left ventricle (solid trace is normal case)
Granzier H L , Labeit S Circulation Research 2004;94:284-295
Copyright American Heart Association
Passive force in cardiac muscle
3-D reconstruction of a typical collagen fibre in a ventricular trabecula fixed
at near-resting sarcomere length
Hanley P J et al. J Physiol 1999;517:831-837
1999 by The Physiological Society
Collagen is furrowed in cardiac muscle
Collagen surrounding skeletal muscle
From Histology and Virtual Microscopy Learning Resources -
http://histology.med.umich.edu/medical/muscle

Enlargement of box here
Collagen surrounding skeletal muscle
From slide #059-3 Imagescope from
Histology and Virtual Microscopy Learning Resources
http://histology.med.umich.edu/medical/muscle

Classical Lumped Parameter
Models
Classical approach to
modeling muscle
Muscle
Tendon
Elastic component has tension
proportional to distortion
T = k Dx
x
x = L
0
+ Dx
Slack length Distortion
Note that distortion applies
only for positive values
Dx > 0
T = 0 for x < L
0

x
Hence no force for lengths less than the slack length
Series elastic component -
not exactly linear but is close enough
Jewell and Wilke, 1958
Add tensions for components
in parallel
T = k
1
Dx
1
+ k
2
Dx
2

x and dx/dt are same for both elements
k
2

k
1

x = x
1
= x
2
Components in series have same
tension but must add lengths
T = k
1
Dx
1
= k
2
Dx
2
=> same for both elements
dx/dt = dx
1
/dt +

dx
2
/dt => add change in
length for two elements
k
2
k
1

x = x
1
+

x
2


x
2
x
1

Applied force
Note that series and parallel depends
on context and not just geometry
Series - if both elements above slack then
k
Total
= 1/(1/k
1
+ 1/k
2
)
k
2
k
1

Applied force
Parallel - if both elements above slack then
k
Total
= k
1
+ k
2

k
2
k
1

Applied force
Active state T
0
and viscous
element B
Tension source generates
tension only (can be time
and length dependent)
Dashpot generates
tension in response to
rate of change in length
i.e. T=B*dx
1
/dt
Active state T
0
and viscous
element B
The activation of skeletal muscle by
single and repetitive stimulation. A
fused tetanus occurs at stimulus
frequency greater than about 10 Hz
Skeletal activation process:
Twitches and tetanus
The activation of skeletal muscle by
by two stimulus pulses at times S1
and S2. As times S1 and S2 get
closer, the twitches fuse.
The activation of skeletal muscle by single and repetitive stimulation. A
fused tetanus occurs at stimulus frequency greater than about 10 Hz
Skeletal activation process:
Twitches and tetanus
Active state during twitch can be
tracked by delayed release of muscle
After release, muscle will shorten until string is taut, and then net force is measured.
Effect of viscosity element is only seen during internal shortening. At peak tension,
dT/dt = 0. We know -dX
1
/dt = dX
2
/dt = 0 so no contribution from viscous element.
Note: muscle is below slack length, so no contribution from K
PE
=> T = T
0.
. Under
these conditions, the total tension is the same as active element.
Each successive trace
corresponds to later
release of muscle
String is slack so no
force is measured
until release of
catch - then some
active shortening
until string is pulled
taut
What is going on at active
element?
Before release, active element will shorten so that x
1
is below slack length.
Then we are left with picture above. After release, muscle will shorten to a
new shorter length where isometric contraction will occur. Only the series
elastic element can change length instantaneously. The active element will
actively shorten more slowly until the T
0
is just balanced by the series elastic
element. As shortening stops, the viscosity element does not contribute.
Now total tension is same as active element. Note that as time continues,
the force in the active element continues to fall and x
1
will increase as the
active element lengthens. Now the viscosity element will again contribute,
but now the viscosity element resist lengthening active element.
Classical approach to
modeling muscle
dT/dt = 0 at peaks and troughs to track T = T
0

Solved problem from McMahon, p. 21
Solved problem from McMahon
Solved problem from McMahon
(Assumes not less than slack length)
the derivative of
^
Solved problem from McMahon
Exponential fall
has terms with
exp(-(t-t
init
)/t)
where t = B/K
Exponential rise
has terms with
(1-exp(-(t-t
init
)/t)
where t = B/K
Method 2
0 at t 0 T
init init

T
0

0
t = 0 C C +A
2C +A t -> infinity
Assume output of active element is T
0
for infinitely long
) e 1 ( T ) C ( T
C
B
K
0


0
T T

t t / ) t t (
init
/ ) t t (
init init
e T ) e 1 ( T ) t ( T

where t = B/K

Therefore and ) e 1 ( T ) t ( T
t
B
K
0


Method 2
T
0

0
t = 0 C C +A
2C +A t -> infinity
) A (
B
K
e ) C ( T ) A C ( T


t t / ) t t (
init
/ ) t t (
init init
e T ) e 1 ( T ) t ( T

where t = B/K

Therefore
C at t ) C ( T T
init init

Now repeat process but move forward in time so t
init
= C
0 T

and
Assume output of active element is 0 for infinitely long
) C ( T
t / ) C t (
e ) C ( T ) t ( T

and
) A C ( T
Note that time constant will change if
more elastic elements are added

For example, if K
PE
and K
SE
are above slack length, then
t = B/(K
PE
+ K
SE
)
Warning: K
PE
and K
SE
are nonlinear elements
Answer to part b
= 1 when C (A can be anything)

t = 0 C C +A
2C +A
T
0

0
t = 0 C C +A
2C +A
= 1 when A (C can be anything)

t = 0 C
C +A 2C +A
= 2 when A and C 0


T
0

0
T
0

0
x =(1+e
K( A+C)/B
)
= 2 when C 0 and A 0