scholarly journals Delayed rectification in the transverse tubules: origin of the late after-potential in frog skeletal muscle.

1977 ◽  
Vol 70 (1) ◽  
pp. 1-21 ◽  
Author(s):  
G E Kirsch ◽  
R A Nichols ◽  
S Nakajima
Keyword(s):  
Skeletal Muscle ◽  
Muscle Fibers ◽  
Tetanic Stimulation ◽  
Potassium Accumulation ◽  
Transverse Tubular System ◽  
System A ◽  
Delayed Rectification ◽  
Wide Range ◽  
Time Courses ◽  
T System

Tetanic stimulation of skeletal muscle fibers elicits a train of spikes followed by a long-lasting depolarization called the late after-potential (LAP). We have conducted experiments to determine the origin of the LAP. Isolated single muscle fibers were treated with a high potassium solution (5 mM or 10 mM K) followed by a sudden reduction of potassium concentration to 2.5 mM. This procedure produced a slow repolarization (K repolarization), which reflects a diffusional outflow of potassium from inside the lumen of the transverse tubular system (T system). Tetanic stimulation was then applied to the same fiber and the LAP was recorded. The time courses of K repolarization and LAP decay were compared and found to be roughly the same. This approximate equality held under various conditions that changed the time courses of both events over a wide range. Both K repolarization and the LAP became slower as fiber radius increased. These results suggest that LAP decay and K repolarization represent the same process. Thus, we conclude that the LAP is caused by potassium accumulation in the T system. A consequence of this conclusion is that delayed rectification channels exist in the T system. A rough estimation suggests that the density of delayed rectification channels is less in the T system than in the surface membrane.

scholarly journals A STUDY OF THE T SYSTEM IN RAT HEART

1970 ◽  
Vol 44 (1) ◽  
pp. 1-19 ◽  
Author(s):  
W. G. Forssmann ◽  
L. Girardier
Keyword(s):  
Rat Heart ◽  
Photometric Method ◽  
Ventricular Muscle ◽  
Transverse Tubular System ◽  
System A ◽  
L System ◽  
Line Region ◽  
Specific Granules ◽  
Sizable Proportion ◽  
T System

The technique of extracellular space tracing with horseradish peroxidase is adapted for labeling the transverse tubular system (T system) in rat heart. In rat ventricular muscle the T system shows extensive branching and remarkable tortuosity. The T system can only be defined operationally, since it does not display specific morphological features throughout its entire structure. Owing to branching of the T system, a sizable proportion of the apposition between the T system and L system (or closed system) occurs at the level of longitudinal branches of the T system and is not restricted to the Z line region. The regions of apposition between the T system and L system are analyzed in rat ventricular muscle and skeletal muscle (diaphragm) and compared with the intercellular tight junctions (nexuses) of heart muscle by the use of a photometric method. The over-all thickness of the nexus is significantly smaller than that of T-L junctions in both cardiac and skeletal muscles. The thickness of the membranes of the T and L systems are not significantly different in the two muscles, but the gap between both membranes is larger in the heart. In atrial muscle the following two types of cells are found: (a) those cells with a well-developed T system in which the tubular diameter is quite uniform and the orientation predominantly longitudinal and, (b) cells with no T system, but with a well-developed L system. Atrial cells possessing a T system are richly provided with specific granules and show little micropinocytotic activity, whereas cells devoid of T system show intense micropinocytotic activity and few specific granules. The possible functional implications of these findings are discussed.

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 On the connection between the transverse tubules and the plasma membrane in frog semitendinosus skeletal muscle

1975 ◽  
Vol 64 (3) ◽  
pp. 734-740 ◽  
Author(s):  
G Zampighi ◽  
J Vergara ◽  
F Ramon
Keyword(s):  
Skeletal Muscle ◽  
Plasma Membrane ◽  
Muscle Fibers ◽  
Indirect Evidence ◽  
Electrical Signal ◽  
Ruthenium Red ◽  
Transitional Region ◽  
Transverse Tubular System ◽  
Pinocytotic Vesicles ◽  
And Function

The transverse tubular system (TTS) of skeletal muscle fibers represents the morphological basis for the inward spread of conduction of the electrical signal that triggers muscle contraction. A historical account of the main steps contributing to the elucidation of the structure and function of the TSS has been presented by Huxley (1971). While the localization of the TSS and its association with the sarcoplasmic reticulum (SR) is well documented; there is still a need further to develop our knowledge of the morphology of the connection between the TSS and the plasma membrane. It is generally believed that the TSS opens directly to the extracellular space and that there is continuity between its membrane and the sarcolemma. However, direct observation of such a connection has been clearly shown only for the myotome of fish (Franzini-Armstrong and Porter, 1964). In other muscle fibers, only indirect evidence of the connection has been provided by experiments showing penetration of extracellular tracers into the TSS. These extracellular markers were also observed inside another membrane-bounded compartment consisting of round profiles named "caveolae" (Yamada, 1955) or "pinocytotic vesicles" (Ashurst, 1969). The present study deals with the communication between the TTS, caveolae, and plasma membrane (Peachey, 1965); Ezerman and Ishikawa, 1967; Schiaffino and Margreth, 1968; and Rayns et al., 1968). A detailed study of the caveolae compartment was undertaken with ruthenium red as an electron-dense tracer. As a result of this study, we propose that in certain species the caveolae compartment represents the transitional region in the connection between the TSS and the sarcolemma.

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 Role of physiological ClC-1 Cl− ion channel regulation for the excitability and function of working skeletal muscle

2016 ◽  
Vol 147 (4) ◽  
pp. 291-308 ◽  
Author(s):  
Thomas Holm Pedersen ◽  
Anders Riisager ◽  
Frank Vincenzo de Paoli ◽  
Tsung-Yu Chen ◽  
Ole Bækgaard Nielsen
Keyword(s):  
Skeletal Muscle ◽  
Muscle Activity ◽  
Muscle Fibers ◽  
Membrane Conductance ◽  
Membrane Properties ◽  
Equilibrium Potential ◽  
Myotonia Congenita ◽  
Clcn1 Gene ◽  
Wide Range ◽  
Muscle Excitability

Electrical membrane properties of skeletal muscle fibers have been thoroughly studied over the last five to six decades. This has shown that muscle fibers from a wide range of species, including fish, amphibians, reptiles, birds, and mammals, are all characterized by high resting membrane permeability for Cl− ions. Thus, in resting human muscle, ClC-1 Cl− ion channels account for ∼80% of the membrane conductance, and because active Cl− transport is limited in muscle fibers, the equilibrium potential for Cl− lies close to the resting membrane potential. These conditions—high membrane conductance and passive distribution—enable ClC-1 to conduct membrane current that inhibits muscle excitability. This depressing effect of ClC-1 current on muscle excitability has mostly been associated with skeletal muscle hyperexcitability in myotonia congenita, which arises from loss-of-function mutations in the CLCN1 gene. However, given that ClC-1 must be drastically inhibited (∼80%) before myotonia develops, more recent studies have explored whether acute and more subtle ClC-1 regulation contributes to controlling the excitability of working muscle. Methods were developed to measure ClC-1 function with subsecond temporal resolution in action potential firing muscle fibers. These and other techniques have revealed that ClC-1 function is controlled by multiple cellular signals during muscle activity. Thus, onset of muscle activity triggers ClC-1 inhibition via protein kinase C, intracellular acidosis, and lactate ions. This inhibition is important for preserving excitability of working muscle in the face of activity-induced elevation of extracellular K+ and accumulating inactivation of voltage-gated sodium channels. Furthermore, during prolonged activity, a marked ClC-1 activation can develop that compromises muscle excitability. Data from ClC-1 expression systems suggest that this ClC-1 activation may arise from loss of regulation by adenosine nucleotides and/or oxidation. The present review summarizes the current knowledge of the physiological factors that control ClC-1 function in active muscle.

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 Voltage Clamp Experiments on Single Muscle Fibers of Rana pipiens

1972 ◽  
Vol 60 (1) ◽  
pp. 1-19 ◽  
Author(s):  
L. E. Moore
Keyword(s):  
Voltage Clamp ◽  
Rana Pipiens ◽  
Muscle Fibers ◽  
Petroleum Jelly ◽  
Single Muscle ◽  
Transverse Tubular System ◽  
Anomalous Rectification ◽  
Outward Currents ◽  
Delayed Rectification ◽  
Tubular System

A voltage clamp for single muscle fibers has been developed. Stability of the system was achieved when an artificial node was created by enclosing a single muscle fiber in a petroleum jelly seal which served as an analogue of the myelin sheath. Typical voltage clamp records were obtained with large inward transient currents followed by a delayed rectification of the outward currents. These currents looked qualitatively similar when the transverse tubular system was destroyed. Errors in current measurement, especially those due to anomalous rectification, are discussed.

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