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Abstract
Stretch-induced force enhancement has been observed in a variety of muscle preparations and on structural levels ranging from
single fibers to in vivo human muscles. It is a well-accepted property of skeletal muscle. However, the mechanism causing force
enhancement has not been elucidated, although the sarcomere-length non-uniformity theory has received wide support. The purpose
of this paper was to re-investigate stretch-induced force enhancement in frog single fibers by testing specific hypotheses arising from
the sarcomere-length non-uniformity theory. Single fibers dissected from frog tibialis anterior (TA) and lumbricals (n ¼ 12 and 22;
respectively) were mounted in an experimental chamber with physiological Ringer’s solution (pH=7.5) between a force transducer
and a servomotor length controller. The tetantic force–length relationship was determined. Isometric reference forces were
determined at optimum length (corresponding to the maximal, active, isometric force), and at the initial and final lengths of the
stretch experiments. Stretch experiments were performed on the descending limb of the force–length relationship after maximal
tetanic force was reached. Stretches of 2.5–10% (TA) and 5–15% lumbricals of fiber length were performed at 0.1–1.5 fiber lengths/s.
The stretch-induced, steady-state, active isometric force was always equal or greater than the purely isometric force at the muscle
length from which the stretch was initiated. Moreover, for stretches of 5% fiber length or greater, and initiated near the optimum
length of the fiber, the stretch-enhanced active force always exceeded the maximal active isometric force at optimum length. Finally,
we observed a stretch-induced enhancement of passive force. We conclude from these results that the sarcomere length non-
uniformity theory alone cannot explain the observed force enhancement, and that part of the force enhancement is associated with a
passive force that is substantially greater after active compared to passive muscle stretch.
r 2003 Elsevier Science Ltd. All rights reserved.
Keywords: Striated muscle; Single fibers; Force enhancement; Muscle mechanics; Force production; Cross-bridge theory; Sarcomere length non-
uniformity theory
1. Introduction than any other theory (Edman et al., 1982; Edman and
Tsuchiya, 1996; Julian and Morgan, 1979b; Julian and
The steady-state isometric force following active Morgan, 1979a; Morgan, 1990; Morgan, 1994; Morgan
muscle stretch is greater than the purely isometric force et al., 2000). The sarcomere length non-uniformity
at the corresponding length. This property of skeletal theory is based on the idea that during stretch, on the
muscle has been observed at structural levels ranging descending limb of the force–length relationship, some
from single fibers (e.g. Edman et al., 1978) to in vivo sarcomeres elongate less than average (and so, can
human skeletal muscle (Lee and Herzog, 2002), and it produce more active force at steady-state than if they
has been termed ‘‘force enhancement following stretch’’ were at the average sarcomere length), while others
(Herzog, 1998). There is no generally accepted explana- elongate more than average and produce most of their
tion for this force enhancement, although the sarcomere force passively (Fig. 1).
length non-uniformity theory has received more support If sarcomere length non-uniformity was the only
mechanism for force enhancement, the steady-state
*Corresponding author. Tel.: +1-403-220-8525; fax: +1-403-284-
force enhancement following stretch should not exceed
3553. the purely isometric force at optimum muscle length.
E-mail address: walter@kin.ucalgary.ca (W. Herzog). However, recently we observed force enhancements of
0021-9290/03/$ - see front matter r 2003 Elsevier Science Ltd. All rights reserved.
doi:10.1016/S0021-9290(03)00155-6
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the corresponding isometric reference contractions. Par- decay following isometric and stretch contractions was
allelism was evaluated by fitting a least-squares straight evaluated by calculating the time from the last activa-
line through the data points for the last 0.5 s of tetanus tion pulse to the instant when force had decreased by
and testing for differences in these slopes (a ¼ 0:05). 50% (T50), and the time from 80% to 20% of the force
For stretching of the muscle fibers, the fibers were just after deactivation (T80–20); and (ii) linear regression
placed at an initial length (Li ), activated to reach at least analyses (a ¼ 0:05) were performed for passive force
95% of the maximal isometric force at that length enhancement as a function of stretch amplitude, passive
(E0.85 s), stretched to the final length (Lf ), and held at force enhancement as a function of total force enhance-
the final length until steady-state force was obtained (as ment, force decay times as a function of stretch
determined in pilot experiments). The initial fiber amplitude, and force enhancement as a function of
lengths were chosen at approximately 5%, 10%, and stretch amplitudes.
15% of fiber lengths beyond the plateau of the force–
length relationship (i.e. on the descending limb).
Stretch amplitudes ranged from 2.5% to 15% of fiber 3. Results
lengths, and the speed of stretch was 10%, 20%, 40%,
50%, 100% or 150% of fiber length/s. One full stretch For fibers from both muscles, and for all stretch
experiment consisted of the following eight tests: amplitudes and stretch speeds considered in this study,
isometric contractions at optimum length, Li ; and Lf ; the steady-state forces following active stretch were
the experimental stretch contraction; the experimental always greater than the purely isometric reference
stretch contraction without muscle activation (passive contractions at the corresponding length (Lf ) (Fig. 2).
stretch), and a repeat of the three initial isometric Similarly, the steady-state forces following active stretch
contractions. Following such a test, and assuming that were always equal or greater than the purely isometric
the isometric reference contractions were steady, the reference contractions at the length from which the
next stretch experiment (new Li ; new amplitude, or new stretch was initiated (Li ) (Figs. 2 and 3). Both these
stretch speed) was performed. results were statistically significant.
In many (but not all) cases, the steady-state forces
2.2. Data analysis and statistics following active stretch were greater than the isometric
reference forces at the plateau of the force–length
All calculations of force enhancement were made for relationship (i.e. at optimum length (Figs. 3a and b
active forces. In order to obtain the active forces for the and 4). Force enhancement above the plateau was likely
isometric contractions, the passive forces just before and to occur when the initial length was close to the plateau
just after activation were joined by a straight line, and (i.e. just slightly on the descending limb of the force–
the force values of this straight line were subtracted length relationship) and when stretch amplitudes were
from the total force. This was done to account for the great (Figs. 5 and 6). Often, force enhancement above
slow (but measurable) decrease in passive force as a
function of time. For the experimental stretch tests,
active and passive fiber stretching was performed for
identical stretch speeds and magnitudes, and the forces
of the passive stretch test were subtracted from the time-
synchronized total forces of the stretch test with the fiber
activated. The active forces recorded at 0.5 s before the
end of contraction, or at 2–3 s (TA) and at 5 s following
the end of the stretch lumbricals, and the residual
passive forces following the active stretch tests recorded
at 1.5 s after deactivation, were used for statistical
analysis. For each initial length investigated in these
experiments, the tetanic and passive force values
following active stretch (i.e. the enhanced forces) were
compared to the isometric reference forces recorded at
the plateau (L0 ), at Li and Lf by means of a one-way
ANOVA for repeated measures. When a significant Fig. 2. Exemplar raw data from a single fiber of TA. Shown are the
difference was observed, contrasts that were chosen a isometric force–time traces at the initial and at two final lengths (initial
plus 5% and initial plus 10% increase in fiber length). Also shown are
priori were used for subsequent mean comparisons. A
four force–time traces for stretches of 2.5%, 5%, 7.5%, and 10% from
significance level of po0:05 was set for all analyses. the initial length at a speed of 0.2 fiber lengths/s. Note that the steady-
Using this initial data analysis, the following post-hoc state force enhancement following all stretches is greater than the
analyses procedures were added: (i) the rate of force purely isometric forces at the final and initial lengths.
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Fig. 4. Exemplar raw data from a single fiber from the lumbricals.
Shown are the isometric force at optimum length (o, top isometric
trace) and the final length (f, bottom isometric trace). The active (s, top
trace) and passive (p, bottom trace) stretch tests are also shown. Note
that force enhancement following the active stretch greatly exceeds the
purely isometric force at optimum fiber length. Observe further that
the passive force in the active stretch experiments (after fiber
deactivation, arrow) is greater than the corresponding passive forces
of the isometric test at the final length and the passive force following
the passive stretch test. We called this additional passive force
following active fiber stretching ‘‘passive force enhancement’’ (see
arrow). Initial length was 5% greater than optimum length and the
stretch amplitude was 15% of optimum fiber length at a speed of 40%
fiber length/s.
Fig. 3. Exemplar raw data from one fiber from the lumbricals. Shown
are force–time traces of three isometric contractions at fiber optimum
length (o, top isometric trace), the initial length (i, mid-isometric trace),
and the final length (f, bottom isometric trace), as well as a stretch test
from the initial to the final fiber length (s). Note the sharp increase in
force at the onset of stretch (the short-range stiffness), followed by a
less steep slope in (a) and (b). Note further that the steady-state force
enhancement is greater than any of the isometric forces, particularly
the optimum isometric force (top isometric trace) in (a) and (b). In (c),
force enhancement does not exceed the optimum isometric force as the
stretch amplitude was not sufficient to achieve this result. Initial
lengths: 5%, 10%, and 15% fiber lengths greater than optimum Fig. 5. Mean force–length relationship and mean (71 S.E.) steady-
lengths for (a), (b), and (c), respectively. Final lengths: 20% greater state total and passive force enhancement following stretch tests in
than optimum length in all three cases, therefore the stretch amplitude lumbrical fibers (n=22). Forces were normalized with respect to the
was 15%, 10%, and 5% of fiber length for (a), (b), and (c). Stretch maximal isometric force for comparison across fibers. Fiber lengths
speed was 40% fiber lengths/s in all cases. were normalized relative to the optimum fiber length; i.e. the length at
which the active isometric force was greatest. Note that the mean force
enhancement for several conditions exceeded the maximal isometric
forces. Note further, that force enhancement was directly related to the
stretch magnitude for all initial lengths.
the plateau was associated with substantial passive
force enhancement following deactivation of the muscle
(Fig. 4). When the passive force enhancement values
were subtracted from the total force enhancement values
for the cases in which force enhancement exceeded the Passive force enhancement was positively correlated
plateau forces, force enhancement values were reduced with the stretch amplitude and the amount of total force
to the isometric plateau values, or below, at least in a enhancement (Figs. 10a and b).
statistical sense (Fig. 7). Finally, the force decay times, T50 and T80–20, were
Force enhancement was found to be positively related typically greater following the experimental stretch
to the magnitude of stretch, as had been shown in tests, compared to the decay times observed for the
previous studies (Figs. 5 and 6) (Edman et al., 1978; isometric contractions at the corresponding muscle
Edman et al., 1982). Interestingly, the amount of force lengths. Furthermore, force decay times tended to
enhancement had a small, but statistically significant, increase, i.e. the average rate of change in force
negative relationship with the stretch rate for TA fibers tended to decrease, with increasing stretch magnitudes
(Fig. 8), but not the lumbrical fibers (Fig. 9). (Fig. 11).
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4. Discussion
Fig. 9. Exemplar raw data of stretch tests at different speeds for a single fiber from the lumbricals. Shown is an isometric contraction at the initial
length (i=5% greater than optimum length) and 5 stretch tests of 15% fiber length at speeds of approximately 20%, 30%, 40%, 100%, and 150%
fiber length/s (20, 30, 40, 100, and 150, respectively). Note that the steady-state forces following all stretches were similar. No significant relationship
between force enhancement and stretch speed was found in lumbrical fibers.
Fig. 10. Passive force enhancement in lumbrical fibers as a function of the stretch amplitude: (a), and the total force enhancement (b). The great
variation in the passive force enhancement for a given stretch amplitude is partly caused by the dependence of passive force enhancement on the
absolute fiber lengths. For example, a stretch of 5% starting at a fiber length of 5% greater than optimum length typically gave a smaller passive force
enhancement than a stretch of equal magnitude starting at a fiber length of 15% greater than optimum.
enhancement above the isometric forces at optimum the plateau forces requires a different or an additional,
length are a true reflection of the mechanisms underlying mechanism.
the force enhancement process, rather than an artefact There are conceptually two possibilities to explain the
associated with the calculation of active force. steady-state forces following active stretch above the
Based on our results, force enhancement following plateau forces: (a) cross-bridge based mechanisms; or (b)
stretch cannot be explained using the sarcomere length non-cross-bridge based mechanisms. If a cross-bridge
non-uniformity theory exclusively. At best, this theory based mechanism was responsible for the force enhance-
can explain force enhancements up to the isometric ment above plateau, then cross-bridges following active
plateau forces. Steady-state force enhancement above stretch should be stronger, or there should be a greater
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D.E. Rassier et al. / Journal of Biomechanics 36 (2003) 1309–1316 1315
possible mechanisms for this decreased rate of force Edman, K.A.P., Elzinga, G., Noble, M.I.M., 1978. Enhancement of
decrease following active fiber stretching compared to mechanical performance by stretch during tetanic contractions
the isometric reference contractions. However, several of vertebrate skeletal muscle fibres. Journal of Physiology 281, 139–
155.
possibilities exist. These include a change in rate of Edman, K.A.P., Elzinga, G., Noble, M.I.M., 1982. Residual force
cross-bridge attachment/detachment, a decreased rate of enhancement after stretch of contracting frog single muscle fibers.
Ca2+ uptake into the sarcoplasmic reticulum, or an Journal of General Physiology 80, 769–784.
increase in a passive viscoelastic response (Brown and Forcinito, M., Epstein, M., Herzog, W., 1998. Can a rheological
Loeb, 2000). Future experiments should be conducted to muscle model predict force depression/enhancement? Journal of
Biomechanics 31, 1093–1099.
investigate this finding in detail. Herzog, W., 1998. History dependence of force production in skeletal
muscle: a proposal for mechanisms. Journal of Electromyography
and Kinesiology 8, 111–117.
5. Summary and conclusion Herzog, W., Leonard, T.R., 2000. The history dependence of force
production in mammaliar skeletal muscle following stretch-short-
ening and shortening-stretch cycles. Journal of Biomechanics 33,
This is the first study to demonstrate that the steady- 531–542.
state force enhancement following active fiber stretching Herzog, W., Leonard, T.R., 2002. Force enhancement following
can give rise to forces that are substantially greater than stretching of skeletal muscle: a new mechanism. Journal of
the isometric forces at optimum (plateau) length. These Experimental Biology 205, 1275–1283.
results eliminate the sarcomere length non-uniformity Huxley, A.F., 1957. Muscle structure and theories of con-
traction. Progress in Biophysics and Biophysical Chemistry 7,
theory as the sole explanation for force enhancement. 255–318.
Furthermore, we observed a passive component of force Huxley, A.F., Simmons, R.M., 1971. Proposed mechanism of force
enhancement that might contribute to the total force generation in striated muscle. Nature 233, 533–538.
enhancement, and might account for the steady-state Julian, F.J., Morgan, D.L., 1979a. Intersarcomere dynamics during
forces following active fiber stretch in excess of the fixed-end tetanic contractions of frog muscle fibres. Journal of
Physiology 293, 365–378.
isometric plateau forces. The details of this passive force Julian, F.J., Morgan, D.L., 1979b. The effects of tension on non-
enhancement are not known. However, the passive force uniform distribution of length changes applied to frog muscle
enhancement increases with increasing stretch magni- fibres. Journal of Physiology 293, 379–392.
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speed of stretch, and it is correlated with the magnitude stretch of electrically and voluntarily activated human adductor
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2000; Edman and Tsuchiya, 1996; Noble, 1992) have Amenitsch, H., Bernstorff, S., Piazzesi, G., 2000. A combined
suggested that the proteins titin or nebulin might engage mechanical and X-ray diffraction study of stretch potentiation
upon active stretching of muscle or muscle fibers, and so in single frog muscle fibres. Journal of Physiology 526 (3),
might produce the passive force enhancement observed 589–596.
Mar!echal, G., Plaghki, L., 1979. The deficit of the isometric tetanic
here for the first time in single fibers. tension redeveloped after a release of frog muscle at a constant
velocity. Journal of General Physiology 73, 453–467.
Morgan, D.L., 1990. New insights into the behavior of muscle during
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