Effects of blebbistatin and Ca2+ concentration on force produced during stretch of skeletal muscle fibers

2010 ◽  
Vol 299 (5) ◽  
pp. C1127-C1135 ◽  
Author(s):  
Fabio C. Minozzo ◽  
Dilson E. Rassier
Keyword(s):  
Skeletal Muscle ◽  
Sarcomere Length ◽  
Muscle Fibers ◽  
Critical Force ◽  
Power Stroke ◽  
Optimal Length ◽  
Maximal Force ◽  
The Cross ◽  
Cross Bridges ◽  
Two Phases

When activated muscle fibers are stretched at low speeds [≤2 optimal length ( Lo)/s], force increases in two phases, marked by a change in slope [critical force (Pc)] that happens at a critical sarcomere length extension ( Lc). Some studies attribute Pc to the number of attached cross bridges before stretch, while others attribute it to cross bridges in a pre-power-stroke state. In this study, we reinvestigated the mechanisms of forces produced during stretch by altering either the number of cross bridges attached to actin or the cross-bridge state before stretch. Two sets of experiments were performed: 1) activated fibers were stretched by 3% Lo at speeds of 1.0, 2.0, and 3.0 Lo/s in different pCa2+ (4.5, 5.0, 5.5, 6.0), or 2) activated fibers were stretched by 3% Lo at 2 Lo/s in pCa2+ 4.5 containing either 5 μM blebbistatin(+/−) or its inactive isomer (+/+). All stretches started at a sarcomere length (SL) of 2.5 μm. When fibers were activated at a pCa2+ of 4.5, Pc was 2.47 ± 0.11 maximal force developed before stretch (Po) and decreased with lower concentrations of Ca2+. Lc was not Ca2+ dependent; the pooled experiments provided a Lc of 14.34 ± 0.34 nm/half-sarcomere (HS). Pc and Lc did not change with velocities of stretch. Fibers activated in blebbistatin(+/−) showed a higher Pc (2.94 ± 0.17 Po) and Lc (16.30 ± 0.38 nm/HS) than control fibers (Pc 2.31 ± 0.08 Po; Lc 14.05 ± 0.63 nm/HS). The results suggest that forces produced during stretch are caused by both the number of cross bridges attached to actin and the cross bridges in a pre-power-stroke state. Such cross bridges are stretched by large amplitudes before detaching from actin and contribute significantly to the force developed during stretch.

scholarly journals Effects of ramp shortening during linear phase of relaxation on [Ca2+]i in intact skeletal muscle fibers

1999 ◽  
Vol 276 (1) ◽  
pp. C152-C160 ◽  
Author(s):  
Yandong Jiang ◽  
Fred J. Julian
Keyword(s):  
Skeletal Muscle ◽  
Thin Filament ◽  
Sarcomere Length ◽  
Muscle Fibers ◽  
Linear Phase ◽  
Fixed Size ◽  
Skeletal Muscle Fibers ◽  
Straight Line ◽  
Sigmoidal Curve ◽  
Cross Bridges

The effects of shortening distance at V u, the unloaded shortening speed, and filament overlap on the amount of extra Ca2+ released during relaxation in muscle, as indicated by the bump area, were studied. Single, intact frog skeletal muscle fibers at 3°C were used. The myoplasmic free Ca2+ concentration ([Ca2+]i) was estimated by using fura 2 salt injected into the myoplasm. Ramps were applied, either at full overlap with different sizes or at varying overlaps with a fixed size, in the linear phase of relaxation. At full overlap, a plot of bump area vs. ramp size was fit by using a sigmoidal curve with one-half of the bump area equal to 25.9 nm. With a fixed ramp size of 100 nm/half-sarcomere, the plot of bump area vs. mean sarcomere length (SLm) was fit by a straight line intersecting the SLm axis at ∼3.5 μm, close to just no overlap. The results suggest that the transition in the distribution of attached cross bridges from the isometric case to one appropriate for unloaded shortening at V u is completed within 50 nm/half-sarcomere and support the view that attached cross bridges in the overlap zone influence the affinity of Ca2+for troponin C in the thin filament.

scholarly journals Backward Movements of Cross-Bridges by Application of Stretch and by Binding of MgADP to Skeletal Muscle Fibers in the Rigor State as Studied by X-Ray Diffraction

1999 ◽  
Vol 76 (4) ◽  
pp. 1770-1783 ◽  
Author(s):  
Yasunori Takezawa ◽  
Duck-Sool Kim ◽  
Masaki Ogino ◽  
Yasunobu Sugimoto ◽  
Takakazu Kobayashi ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Muscle Fibers ◽  
X Ray Diffraction ◽  
X Ray ◽  
Rigor State ◽  
Skeletal Muscle Fibers ◽  
Cross Bridges

scholarly journals Sarcomere-Length Dependence of the Low Angle X-Ray Pattern from Skeletal Muscle Fibers at Rest and during Isometric Contraction

2012 ◽  
Vol 102 (3) ◽  
pp. 147a-148a
Author(s):  
Gabriella Piazzesi ◽  
Massimo Reconditi ◽  
Elisabetta Brunello ◽  
Luca Fusi ◽  
Marco Linari ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Isometric Contraction ◽  
Sarcomere Length ◽  
Muscle Fibers ◽  
X Ray ◽  
Length Dependence ◽  
Skeletal Muscle Fibers

scholarly journals Calcium-gated calcium channels in sarcoplasmic reticulum of rabbit skinned skeletal muscle fibers.

1986 ◽  
Vol 87 (2) ◽  
pp. 289-303 ◽  
Author(s):  
P Volpe ◽  
G Salviati ◽  
A Chu
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Isometric Force ◽  
Muscle Fibers ◽  
Ruthenium Red ◽  
Scattering Method ◽  
Tension Development ◽  
Maximal Force ◽  
Skeletal Muscle Fibers ◽  
Ca2 Channels

The action of ruthenium red (RR) on Ca2+ loading by and Ca2+ release from the sarcoplasmic reticulum (SR) of chemically skinned skeletal muscle fibers of the rabbit was investigated. Ca2+ loading, in the presence of the precipitating anion pyrophosphate, was monitored by a light-scattering method. Ca2+ release was indirectly measured by following tension development evoked by caffeine. Stimulation of the Ca2+ loading rate by 5 microM RR was dependent on free Ca2+, being maximal at pCa 5.56. Isometric force development induced by 5 mM caffeine was reversibly antagonized by RR. IC50 for the rate of tension rise was 0.5 microM; that for the extent of tension was 4 microM. RR slightly shifted the steady state isometric force/pCa curve toward lower pCa values. At 5 microM RR, the pCa required for half-maximal force was 0.2 log units lower than that of the control, and maximal force was depressed by approximately 16%. These results suggest that RR inhibited Ca2+ release from the SR and stimulated Ca2+ loading into the SR by closing Ca2+-gated Ca2+ channels. Previous studies on isolated SR have indicated the selective presence of such channels in junctional terminal cisternae.

Differential Effects of Sevoflurane, Isoflurane, and Halothane on Ca (2+) Release from the Sarcoplasmic Reticulum of Skeletal Muscle 

1999 ◽  
Vol 91 (1) ◽  
pp. 179-186 ◽  
Author(s):  
Gudrun Kunst ◽  
Bernhard M. Graf ◽  
Rupert Schreiner ◽  
Eike Martin ◽  
Rainer H. A. Fink
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Muscle Fibers ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Maximal Force ◽  
Skinned Muscle ◽  
Skeletal Muscle Fibers ◽  
Force Relation ◽  
The Individual

Background Although malignant hyperthermia after application of sevoflurane has been reported, little is known about its action on intracellular calcium homeostasis of skeletal muscle. The authors compared the effect of sevoflurane with that of isoflurane and halothane on Ca2+ release of mammalian sarcoplasmic reticulum and applied a novel method to quantify Ca2+ turnover in permeabilized skeletal muscle fibers. Methods Liquid sevoflurane, isoflurane, and halothane at 0.6 mM, 3.5 mM, and 7.6 mm were diluted either in weakly calcium buffered solutions with no added Ca2+ (to monitor Ca2+ release) or in strongly Ca2+ buffered solutions with [Ca2+] values between 3 nM and 24.9 microm for [Ca+]-force relations. Measurements were taken on single saponin skinned muscle fiber preparations of BALB/c mice. Individual [Ca2+]force relations were characterized by the Ca2+ concentration at half-maximal force that indicates the sensitivity of the contractile proteins and by the steepness. Each force transient was transformed directly into a Ca2+ transient with respect to the individual [Ca2+]-force relation of the fiber. Results At 0.6 mM, single force transients induced by sevoflurane were lower compared with equimolar concentrations of isoflurane and halothane (P < 0.05). Similarly, calculated peak Ca2+ transients of sevoflurane were lower than those induced by equimolar halothane (P < 0.05). The Ca2+ concentrations at half maximal force were decreased after the addition of sevoflurane, isoflurane, and halothane in a concentration-dependent manner (P < 0.05). Conclusion Whereas sevoflurane, isoflurane, and halothane similarly increase the Ca2+ sensitivity of the contractile apparatus in skeletal muscle fibers, 0.6 mM sevoflurane induces smaller Ca2+ releases from the sarcoplasmic reticulum than does equimolar halothane.

scholarly journals Polarization of Tryptophan Fluorescence from Single Striated Muscle Fibers

1972 ◽  
Vol 59 (1) ◽  
pp. 103-120 ◽  
Author(s):  
C. G. dos Remedios ◽  
R. G. C. Millikan ◽  
M. F. Morales
Keyword(s):  
Sarcomere Length ◽  
Striated Muscle ◽  
Muscle Fibers ◽  
Tryptophan Fluorescence ◽  
Thin Filaments ◽  
A Value ◽  
Striated Muscle Fibers ◽  
Types Of Muscle Fibers ◽  
Cross Bridges ◽  
Contraction And Relaxation

Instrumentation has been developed to detect rapidly the polarization of tryptophan fluorescence from single muscle fibers in rigor, relaxation, and contraction. The polarization parameter (P⊥) obtained by exiciting the muscle tryptophans with light polarized perpendicular to the long axis of the muscle fiber had a magnitude P⊥ (relaxation) > P⊥ (contraction) > P⊥ (rigor) for the three types of muscle fibers examined (glycerinated rabbit psoas, glycerinated dorsal longitudinal flight muscle of Lethocerus americanus, and live semitendinosus of Rana pipiens). P⊥ from single psoas fibers in rigor was found to increase as the sarcomere length increased but in relaxed fibers P⊥ was independent of sarcomere length. After rigor, pyrophosphate produced little or no change in P⊥, but following an adenosine triphosphate (ATP)-containing solution, pyrophosphate produced a value of P⊥ that fell between the contraction and relaxation values. Sinusoidal or square wave oscillations of the muscle of amplitude 0.5–2.0% of the sarcomere length and frequency 1, 2, or 5 Hz were applied in rigor when the myosin cross-bridges are considered to be firmly attached to the thin filaments. No significant changes in P⊥ were observed in either rigor or relaxation. The preceding results together with our present knowledge of tryptophan distribution in the contractile proteins has led us to the conclusion that the parameter P⊥ is a probe of the contractile state of myosin which is probably sensitive to the orientation of the myosin S1 subfragment.

Fall in intracellular Po 2 at the onset of contractions in Xenopus single skeletal muscle fibers

2001 ◽  
Vol 90 (5) ◽  
pp. 1871-1876 ◽  
Author(s):  
Michael C. Hogan
Keyword(s):  
Skeletal Muscle ◽  
Muscle Fibers ◽  
Ringer Solution ◽  
Work Period ◽  
Force Development ◽  
Delayed Onset ◽  
Maximal Force ◽  
Skeletal Muscle Fibers ◽  
Lumbrical Muscle ◽  
Kinetics Of

It remains uncertain whether the delayed onset of mitochondrial respiration on initiation of muscle contractions is related to O2 availability. The purpose of this research was to measure the kinetics of the fall in intracellular Po 2 at the onset of a contractile work period in rested and previously worked single skeletal muscle fibers. Intact single skeletal muscle fibers ( n = 11) from Xenopus laevis were dissected from the lumbrical muscle, injected with an O2-sensitive probe, mounted in a glass chamber, and perfused with Ringer solution (Po 2 = 32 ± 4 Torr and pH = 7.0) at 20°C. Intracellular Po 2 was measured in each fiber during a protocol consisting sequentially of 1-min rest; 3 min of tetanic contractions (1 contraction/2 s); 5-min rest; and, finally, a second 3-min contractile period identical to the first. Maximal force development and the fall in force (to 83 ± 2 vs. 86 ± 3% of maximal force development) in contractile periods 1 and 2, respectively, were not significantly different. The time delay (time before intracellular Po 2 began to decrease after the onset of contractions) was significantly greater ( P < 0.01) in the first contractile period (13 ± 3 s) compared with the second (5 ± 2 s), as was the time to reach 50% of the contractile steady-state intracellular Po 2(28 ± 5 vs. 18 ± 4 s, respectively). In Xenopus single skeletal muscle fibers, 1) the lengthy response time for the fall in intracellular Po 2 at the onset of contractions suggests that intracellular factors other than O2 availability determine the on-kinetics of oxidative phosphorylation and 2) a prior contractile period results in more rapid on-kinetics.

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