scholarly journals The Capacitance of Skeletal Muscle Fibers in Solutions of Low Ionic Strength

1972 ◽  
Vol 59 (3) ◽  
pp. 347-359 ◽  
Author(s):  
P. C. Vaughan ◽  
J. N. Howell ◽  
R. S. Eisenberg
Keyword(s):  
Skeletal Muscle ◽  
Ionic Strength ◽  
Surface Membrane ◽  
Muscle Fibers ◽  
External Solution ◽  
Rectangular Pulse ◽  
Bathing Solution ◽  
Skeletal Muscle Fibers ◽  
Tubular System ◽  
Low Ionic Strength

The capacitance of skeletal muscle fibers was measured by recording with one microelectrode the voltage produced by a rectangular pulse of current applied with another microelectrode. The ionic strength of the bathing solution was varied by isosmotic replacement of NaCl with sucrose, the [K] [Cl] product being held constant. The capacitance decreased with decreasing ionic strength, reaching a value of some 2 µF/cm2 in solutions of 30 mM ionic strength, and not decreasing further in solutions of 15 mM ionic strength. The capacitance of glycerol-treated fibers did not change with ionic strength and was also some 2 µF/cm2. It seems likely that lowering the ionic strength reduces the capacitance of the tubular system (defined as the charge stored in the tubular system), and that the 2 µF/cm2 which is insensitive to ionic strength is associated with the surface membrane. The tubular system is open to the external solution in low ionic strength solutions since peroxidase is able to diffuse into the lumen of the tubules. Twitches and action potentials were also recorded from fibers in low ionic strength solutions, even though the capacitance of the tubular system was very small in these solutions. This finding can be explained if there is an action potential—like mechanism in the tubular membrane.

scholarly journals Block of single L-type Ca2+ channels in skeletal muscle fibers by aminoglycoside antibiotics.

1996 ◽  
Vol 107 (3) ◽  
pp. 421-432 ◽  
Author(s):  
C M Haws ◽  
B D Winegar ◽  
J B Lansman
Keyword(s):  
Skeletal Muscle ◽  
Single Channel ◽  
Muscle Fibers ◽  
External Solution ◽  
Bimolecular Reaction ◽  
Aminoglycoside Antibiotics ◽  
Ion Concentration ◽  
Ion Flow ◽  
Skeletal Muscle Fibers ◽  
Ca2 Channels

The activity of single L-type Ca2+ channels was recorded from cell-attached patches on acutely isolated skeletal muscle fibers from the mouse. The experiments were concerned with the mechanism by which aminoglycoside antibiotics inhibit ion flow through the channel. Aminoglycosides produced discrete fluctuations in the single-channel current when added to the external solution. The blocking kinetics could be described as a simple bimolecular reaction between an aminoglycoside molecule and the open channel. The blocking rate was found to be increased when either the membrane potential was made more negative or the concentration of external permeant ion was reduced. Both of these effects are consistent with a blocking site that is located within the channel pore. Other features of block, however, were incompatible with a simple pore blocking mechanism. Hyperpolarization enhanced the rate of unblocking, even though an aminoglycoside molecule must dissociate from its binding site in the channel toward the external solution against the membrane field. Raising the external permeant ion concentration also enhanced the rate of unblocking. This latter finding suggests that aminglycoside affinity is modified by repulsive interactions that arise when the pore is simultaneously occupied by a permeant ion and an aminoglycoside molecule.

scholarly journals Optical signals from surface and T system membranes in skeletal muscle fibers. Experiments with the potentiometric dye NK2367.

1982 ◽  
Vol 80 (2) ◽  
pp. 203-230 ◽  
Author(s):  
J A Heiny ◽  
J Vergara
Keyword(s):  
Skeletal Muscle ◽  
Voltage Clamp ◽  
Surface Membrane ◽  
Muscle Fibers ◽  
Optical Signal ◽  
Intensity Change ◽  
Optical Signals ◽  
Potential Control ◽  
Skeletal Muscle Fibers ◽  
T System

Absorbance signals were recorded from cut single skeletal muscle fibers stained with the nonpenetrating potentiometric dye NK2367 and mounted in a three-vaseline-gap voltage clamp. The characteristics of the optical signals recorded under current and voltage-clamp conditions were studied at various wavelengths between 500 and 800 nm using unpolarized light. Our results indicate that the absorbance signals recorded with this dye reflect potential changes across both the surface and T system membranes and that the relative contribution of each of these membrane compartments to the total optical change is strongly wavelength dependent. A peak intensity change was detected at 720 nm for the surface membrane signal and at 670 nm for the T system. Evidence for this wavelength-dependent separation derives from an analysis of the kinetics and voltage dependence of the optical signals at different wavelengths, and results obtained in detubulated fibers. The 670-nm optical signal was used to demonstrate the lack of potential control in the T system by the voltage clamp and the effect of a tetrodotoxin (TTX)-sensitive sodium conductance on tubular depolarization.

scholarly journals Isotonic contraction of skinned muscle fibers on a slow time base: effects of ionic strength and calcium.

1981 ◽  
Vol 78 (3) ◽  
pp. 233-257 ◽  
Author(s):  
J Gulati ◽  
R J Podolsky
Keyword(s):  
Ionic Strength ◽  
Initial Velocity ◽  
Muscle Fibers ◽  
Time Base ◽  
Bathing Solution ◽  
Slow Time ◽  
Relative Load ◽  
Residual Tension ◽  
High Calcium ◽  
Low Ionic Strength

The force development by calcium-activated skinned frog skeletal muscle fibers and the motion on a slow time base after a quick decrease in load were studied at 0-1 degrees C as a function of the ionic strength and the degree of activation. The ionic strength was varied between 50 and 190 mM by adding appropriate concentrations of KCl to the bathing solution. Under these conditions, the fibers could be maximally activated for several cycles at low ionic strength without developing residual tension. We found that the steady isometric force in fully activated fibers linearly decreased when the KCl concentration was increased from 0 to 140 mM. The steady isotonic motion at a given relative load in fully activated fibers was almost the same at KCl concentration greater than or equal to 50 mM. In 0 and 20 mM KCl, the isotonic velocity decreased continuously for more than 300 ms. At a given relative load, the initial velocity of the motion in 0 and 20 mM KCl was about 0.6 and 0.9 times, respectively, that in 140 mM KCl. The initial velocity decreased further when residual tension developed; this observation provides additional evidence that residual tension may reflect the presence of an internal load. The effect of calcium on the motion was examined at 70 mM KCl. In this solution, the motion during the velocity transient at a given relative load appeared to be the same at different levels of activation. The speed of the subsequent motion was almost steady at high calcium levels but decreased continuously in low calcium levels. These results support the idea that at low ionic strength the response of the fiber to calcium is switch-like, but that other factors also affect the contraction mechanism under these conditions.

scholarly journals Chloride currents from the transverse tubular system in adult mammalian skeletal muscle fibers

2010 ◽  
Vol 137 (1) ◽  
pp. 21-41 ◽  
Author(s):  
Marino DiFranco ◽  
Alvaro Herrera ◽  
Julio L. Vergara
Keyword(s):  
Skeletal Muscle ◽  
Time Course ◽  
Muscle Fibers ◽  
Optical Data ◽  
Mammalian Skeletal Muscle ◽  
Chloride Currents ◽  
Actual Distribution ◽  
Transverse Tubular System ◽  
Skeletal Muscle Fibers ◽  
Tubular System

Chloride fluxes are the main contributors to the resting conductance of mammalian skeletal muscle fibers. ClC-1, the most abundant chloride channel isoform in this preparation, is believed to be responsible for this conductance. However, the actual distribution of ClC-1 channels between the surface and transverse tubular system (TTS) membranes has not been assessed in intact muscle fibers. To investigate this issue, we voltageclamped enzymatically dissociated short fibers using a two-microelectrode configuration and simultaneously recorded chloride currents (ICl) and di-8-ANEPPS fluorescence signals to assess membrane potential changes in the TTS. Experiments were conducted in conditions that blocked all but the chloride conductance. Fibers were equilibrated with 40 or 70 mM intracellular chloride to enhance the magnitude of inward ICl, and the specific ClC-1 blocker 9-ACA was used to eliminate these currents whenever necessary. Voltage-dependent di-8-ANEPPS signals and ICl acquired before (control) and after the addition of 9-ACA were comparatively assessed. Early after the onset of stimulus pulses, di-8-ANEPPS signals under control conditions were smaller than those recorded in the presence of 9-ACA. We defined as attenuation the normalized time-dependent difference between these signals. Attenuation was discovered to be ICl dependent since its magnitude varied in close correlation with the amplitude and time course of ICl. While the properties of ICl, and those of the attenuation seen in optical records, could be simultaneously predicted by model simulations when the chloride permeability (PCl) at the surface and TTS membranes were approximately equal, the model failed to explain the optical data if PCl was precluded from the TTS membranes. Since the ratio between the areas of TTS membranes and the sarcolemma is large in mammalian muscle fibers, our results demonstrate that a significant fraction of the experimentally recorded ICl arises from TTS contributions.

scholarly journals An improved vaseline gap voltage clamp for skeletal muscle fibers.

1976 ◽  
Vol 67 (3) ◽  
pp. 265-293 ◽  
Author(s):  
B Hille ◽  
D T Campbell
Keyword(s):  
Skeletal Muscle ◽  
Voltage Clamp ◽  
Sodium Current ◽  
Muscle Fibers ◽  
Complex Impedance ◽  
Sodium Currents ◽  
Disk Model ◽  
Transverse Tubular System ◽  
Skeletal Muscle Fibers ◽  
Tubular System

A Vaseline gap potentiometric recording and voltage clamp method is developed for frog skeletal muscle fibers. The method is based on the Frankenhaeuser-Dodge voltage clamp for myelinated nerve with modifications to improve the frequency response, to compensate for external series resistance, and to compensate for the complex impedance of the current-passing pathway. Fragments of single muscle fibers are plucked from the semitendinosus muscle and mounted while depolarized by a solution like CsF. After Vaseline seals are formed between fluid pools, the fiber ends are cut once again, the central region is rinsed with Ringer solution, and the feedback amplifiers are turned on. Errors in the potential and current records are assessed by direct measurements with microelectrodes. The passive properties of the preparation are simulated by the "disk" equivalent circuit for the transverse tubular system and the derived parameters are similar to previous measurements with microelectrodes. Action potentials at 5 degrees C are long because of the absence of delayed rectification. Their shape is approximately simulated by solving the disk model with sodium permeability in the surface and tubular membranes. Voltage clamp currents consist primarily of capacity currents and sodium currents. The peak inward sodium current density at 5 degrees C is 3.7 mA/cm2. At 5 degrees C the sodium currents are smoothly graded with increasing depolarization and free of notches suggesting good control of the surface membrane. At higher temperatures a small, late extra inward current appears for small depolarizations that has the properties expected for excitation in the transverse tubular system. Comparison of recorded currents with simulations shows that while the transverse tubular system has regenerative sodium currents, they are too small to make important errors in the total current recorded at the surface under voltage clamp at low temperature. The tubules are definitely not under voltage clamp control.

scholarly journals Ionic Strength and the Contraction Kinetics of Skinned Muscle Fibers

1974 ◽  
Vol 63 (4) ◽  
pp. 509-530 ◽  
Author(s):  
Marc D. Thames ◽  
Louis E. Teichholz ◽  
Richard J. Podolsky
Keyword(s):  
Ionic Strength ◽  
Muscle Fibers ◽  
Bathing Solution ◽  
Shortening Velocity ◽  
Skinned Muscle ◽  
Cross Bridges ◽  
Low Ionic Strength ◽  
Contraction Cycle ◽  
Kinetics Of ◽  
Contraction Kinetics

The influence of KCl concentration on the contraction kinetics of skinned frog muscle fibers at 5–7°C was studied at various calcium levels. The magnitude of the calcium-activated force decreased continuously as the KCl concentration of the bathing solution was increased from 0 to 280 mM. The shortening velocity at a given relative load was unaffected by the level of calcium activation at 140 mM KCl, as has been previously reported by Podolsky and Teichholz (1970. J. Physiol. [Lond.]. 211: 19), and was independent of ionic strength when the KCl concentration was increased from 140 to 280 mM. In contrast, the shortening velocity decreased as the KCl concentration was reduced below 140 mM; the decrease in velocity was enhanced when the fibers were only partially activated. In the low KCl range, the resting tension of the fibers increased after the first contraction cycle. The results suggest that in fibers activated at low ionic strength some of the cross bridges that are formed are abnormal in the sense that they retard shortening and persist in relaxing solution.

scholarly journals Involvement of TRPC in the abnormal calcium influx observed in dystrophic (mdx) mouse skeletal muscle fibers

2002 ◽  
Vol 158 (6) ◽  
pp. 1089-1096 ◽  
Author(s):  
Clarisse Vandebrouck ◽  
Dominique Martin ◽  
Monique Colson-Van Schoor ◽  
Huguette Debaix ◽  
Philippe Gailly
Keyword(s):  
Skeletal Muscle ◽  
Transient Receptor Potential ◽  
Calcium Influx ◽  
Surface Membrane ◽  
Muscle Fibers ◽  
Mdx Mouse ◽  
Mdx Mice ◽  
Patch Clamp Technique ◽  
Adult Skeletal Muscle ◽  
Skeletal Muscle Fibers

Duchenne muscular dystrophy results from the lack of dystrophin, a cytoskeletal protein associated with the inner surface membrane, in skeletal muscle. The absence of dystrophin induces an abnormal increase of sarcolemmal calcium influx through cationic channels in adult skeletal muscle fibers from dystrophic (mdx) mice. We observed that the activity of these channels was increased after depletion of the stores of calcium with thapsigargin or caffeine. By analogy with the situation observed in nonexcitable cells, we therefore hypothesized that these store-operated channels could belong to the transient receptor potential channel (TRPC) family. We measured the expression of TRPC isoforms in normal and mdx adult skeletal muscles fibers, and among the seven known isoforms, five were detected (TRPC1, 2, 3, 4, and 6) by RT-PCR. Western blot analysis and immunocytochemistry of normal and mdx muscle fibers demonstrated the localization of TRPC1, 4, and 6 proteins at the plasma membrane. Therefore, an antisense strategy was used to repress these TRPC isoforms. In parallel with the repression of the TRPCs, we observed that the occurrence of calcium leak channels was decreased to one tenth of its control value (patch-clamp technique), showing the involvement of TRPC in the abnormal calcium influx observed in dystrophic fibers.

Involvement of calpains in Ca2+-induced disruption of excitation-contraction coupling in mammalian skeletal muscle fibers

2009 ◽  
Vol 296 (5) ◽  
pp. C1115-C1122 ◽  
Author(s):  
Esther Verburg ◽  
Robyn M. Murphy ◽  
Isabelle Richard ◽  
Graham D. Lamb
Keyword(s):  
Skeletal Muscle ◽  
Surface Membrane ◽  
Muscle Fibers ◽  
Mammalian Skeletal Muscle ◽  
Skinned Muscle ◽  
Ec Coupling ◽  
Calpain 3 ◽  
Skeletal Muscle Fibers ◽  
Longus Muscle ◽  
Rate Of Decline

In skeletal muscle fibers, the coupling between excitation of the surface membrane and the release of Ca2+ from the sarcoplasmic reticulum is irreversibly disrupted if cytoplasmic Ca2+ concentration ([Ca2+]) is raised to micromolar levels for a prolonged period. This excitation-contraction (EC) uncoupling may contribute to muscle weakness after some types of exercise and in certain muscle diseases and has been linked to structural alteration of the triad junctions, but its molecular basis is unclear. Both μ-calpain, a ubiquitous Ca2+-activated protease, and muscle-specific calpain-3 become autolytically activated at micromolar Ca2+ and have been suggested to be responsible for the uncoupling. This study used controlled Ca2+ exposure in mechanically skinned fibers from extensor digitorum longus muscle to show that EC uncoupling still occurs in muscle fibers of calpain-3-deficient mice, with a Ca2+ dependence indistinguishable from that in normal mice and rats. Western blotting of muscle fibers that had been partially EC uncoupled by exposure to an intermediate Ca2+ level (∼5 μM Ca2+ for 3 min, no ATP) showed the presence of autolytic activation of a proportion of the μ-calpain present, but with little or no activation of calpain-3. Homogenates of normal and calpain-3-deficient muscles exposed to micromolar Ca2+ displayed similar levels of diffusible proteolytic activity, as gauged by the rate of decline of passive force in stretched, skinned muscle fibers. Exogenously added μ-calpain, preactivated by elevated [Ca2+] and applied in the presence of 1 μM Ca2+, disrupted EC coupling in a manner similar to raised [Ca2+]. We conclude that calpain-3 is not responsible for Ca2+-induced disruption of EC coupling, but that μ-calpain is a plausible candidate.

Sign in / Sign up

Export Citation Format

Share Document