STUDIES OF THE TRIAD I. Structure of the Junction in Frog Twitch Fibers
TL;DR: The structure of the junction between sarcoplasmic reticulum (SR) and transverse tubular (T) system at the triad has been studied in twitch fibers of the frog and the junctional area between the feet is apparently empty.
Abstract: The structure of the junction between sarcoplasmic reticulum (SR) and transverse tubular (T) system at the triad has been studied in twitch fibers of the frog. The junction is formed by flattened surfaces of the SR lateral sacs and the T-system tubule, which face each other at a distance of 120–140 A. At periodic intervals of about 300 A, the SR membrane forms small projections, whose tips are joined to the T system membrane by some amorphous material. The SR projections and the amorphous material are here called SR feet. The feet are disposed in two parallel rows, two such rows being present on either side of the T-system tubule. The junctional area between the feet is apparently empty. The feet cover no more than 30% of the T system surface area and 3% of the total SR area. The functional significance of this interpretation of the junctional structure is discussed.
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TL;DR: It appears likely that this condition is associated with and likely caused by muscle injury, such that the SR releases less Ca2+ at low frequencies of activation, and LFF could result from a reduced membrane excitability,such that the sarcolemma action potential frequency is considerably less than the stimulation frequency.
Abstract: Fatigue, defined as the failure to maintain the required or expected power output, is a complex problem, since multiple factors are clearly involved, with the relative importance of each dependent on the fiber type composition of the contracting muscles(s), and the intensity, type, and duration of the contractile activity. The primary sites of fatigue appear to be within the muscle cell itself and for the most part do not involve the central nervous system or the neuromuscular junction. The major hypotheses of fatigue center on disturbances in the surface membrane, E-C coupling, or metabolic events. The cell sites most frequently linked to the etiology of skeletal muscle fatigue are shown in Figure 1. Skeletal muscles are composed of at least four distinct fiber types (3 fast twitch and 1 slow twitch), with the slow type I and fast type IIa fibers containing the highest mitochondrial content and fatigue resistance. Despite fiber type differences in the degree of fatigability, the contractile properties undergo characteristic changes with the development of fatigue that can be observed in whole muscles, single motor units, and single fibers. The Po declines, and the contraction and relaxation times are prolonged. Additionally, there is a decrease in the peak rate of tension development and decline and a reduced Vo. Changes in Vo are more resistant to fatigue than Po and are not observed until Po has declined by at least 10% of its initial prefatigued value. However, the reduced peak power by which fatigue is defined results from both a reduction in Vo and Po. In the absence of muscle fiber damage, the prolonged relaxation time associated with fatigue causes the force-frequency curve to shift to the left, such that peak tensions are obtained at lower frequencies of stimulation. In a mechanism not clearly understood, the central nervous system senses this condition and reduces the alpha-motor nerve activation frequency as fatigue develops. In some cases, selective LFF develops that displaces the force-frequency curve to the right. Although not proven, it appears likely that this condition is associated with and likely caused by muscle injury, such that the SR releases less Ca2+ at low frequencies of activation. Alternatively, LFF could result from a reduced membrane excitability, such that the sarcolemma action potential frequency is considerably less than the stimulation frequency.(ABSTRACT TRUNCATED AT 400 WORDS)
1,531 citations
TL;DR: Controversies surrounding the complex Ca2+ regulation of single RyR channels receive particular attention here and a large body of information is synthesized into a focused perspective of singleRyR channel function.
Abstract: The ryanodine receptors (RyRs) are a family of Ca2+ release channels found on intracellular Ca2+ storage/release organelles. The RyR channels are ubiquitously expressed in many types of cells and participate in a variety of important Ca2+ signaling phenomena (neurotransmission, secretion, etc.). In striated muscle, the RyR channels represent the primary pathway for Ca2+ release during the excitation-contraction coupling process. In general, the signals that activate the RyR channels are known (e.g., sarcolemmal Ca2+ influx or depolarization), but the specific mechanisms involved are still being debated. The signals that modulate and/or turn off the RyR channels remain ambiguous and the mechanisms involved unclear. Over the last decade, studies of RyR-mediated Ca2+ release have taken many forms and have steadily advanced our knowledge. This robust field, however, is not without controversial ideas and contradictory results. Controversies surrounding the complex Ca2+ regulation of single RyR channels receive particular attention here. In addition, a large body of information is synthesized into a focused perspective of single RyR channel function. The present status of the single RyR channel field and its likely future directions are also discussed.
1,075 citations
TL;DR: The predicted structure suggests that the calcium release channel activity resides in the C-terminal region of the receptor molecule, whereas the remaining portion constitutes the 'foot' structure spanning the junctional gap between the sarcoplasmic reticulum and the transverse tubule.
Abstract: The sequence of 5,037 amino acids composing the ryanodine receptor from rabbit skeletal muscle sarcoplasmic reticulum has been deduced by cloning and sequencing the complementary DNA The predicted structure suggests that the calcium release channel activity resides in the C-terminal region of the receptor molecule, whereas the remaining portion constitutes the 'foot' structure spanning the junctional gap between the sarcoplasmic reticulum and the transverse tubule
1,056 citations
TL;DR: It is suggested that a link in excitation-contraction coupling involves the movement of a fixed amount of charge free to move between different locations across the membrane.
Abstract: It is suggested that a link in excitation-contraction coupling involves the movement of a fixed amount of charge free to move between different locations across the membrane.
927 citations
TL;DR: Ca2+ release from the SR induced by T-system depolarization during excitation-contraction coupling in muscle may be effected through a direct association of the T- system with SR Ca22+-release channels.
Abstract: The calcium release channel from rabbit muscle sarcoplasmic reticulum (SR) has been purified and reconstituted as a functional unit in lipid bilayers. Electron microscopy reveals the four-leaf clover structure previously described for the 'feet' that span the transverse tubule (T)-SR junction. Ca2+ release from the SR induced by T-system depolarization during excitation-contraction coupling in muscle may thus be effected through a direct association of the T-system with SR Ca2+-release channels.
881 citations
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TL;DR: This communication reports the use of a commercially available lead citratO to eliminate the lead citrate stain in electron microscopy.
Abstract: A disadvantage of the lead strains used in electron microscopy is the amount of time required for their preparation. One of the more stable and reliable stains, commonly called the lead citrate stain or Reynolds' stain (Reynolds, 1963), is made by mixing lead nitrate and sodium citrate in distilled water, allowing time for lead citrate to form, then adding sodium hydroxide to raise the pH of the solution to 12. This communication reports the use of a commercially available lead citratO to eliminate the
4,496 citations
1,680 citations
TL;DR: A new technique, based on the observations of Doggenweiler and Frenks, wMch has allowed us to demonstrate the presence of similar structures in sections of mouse heart and of mouse liver, and it seems likely that, as lanthanum solutions are brought to a high pH, a colloidal compound is formed which permeates the extracellular space as a tracer.
1,314 citations
TL;DR: Several types of striated muscle have been examined by the technics of electron microscopy and the findings in myotome fibers of Amblystoma larvae, the sartorius, and cardiac muscle of the rat are reported on in some detail.
Abstract: Several types of striated muscle have been examined by the technics of electron microscopy and the findings in myotome fibers of Amblystoma larvae, the sartorius, and cardiac muscle of the rat are reported on in some detail. Particular attention has been given to structural components of the interfibrillar sarcoplasm and most especially to a finely divided, vacuolar system known as the sarcoplasmic reticulum. This consists of membrane-limited vesicles, tubules, and cisternae associated in a continuous reticular structure which forms lace-like sleeves around the myofibrils. It shows a definable organization which repeats with each sarcomere of the fiber so that the entire system is segmented in phase with the striations of the associated myofibrils. Details of these repetitive patterns are presented diagrammatically in Text-figs. 1, 2, and 3 on pages 279, 283, and 288 respectively. The system is continuous across the fiber at the H band level and largely discontinuous longitudinally because of interruptions in the structure at the I and Z band levels. The structure of the system relates it to the endoplasmic reticulum of other cell types. The precise morphological relation of the reticulum to the myofibrils, with specializations opposite the different bands, prompts the supposition that the system is functionally important in muscle contraction. In this regard it is proposed that the membrane limiting the system is polarized like the sarcolemma and that the corresponding potential difference is utilized in the intracellular distribution of the excitatory impulse.
681 citations
TL;DR: The sarcoplasmic reticulum of the frog's sartorius muscle was examined by electron microscopy following sequential fixation in glutaraldehyde and osmium tetroxide and embedding in Epon, finding evidence for continuity of the transverse tubules with the fiber surface.
Abstract: The sarcoplasmic reticulum of the frog's sartorius muscle was examined by electron microscopy following sequential fixation in glutaraldehyde and osmium tetroxide and embedding in Epon. The earlier results of Porter and Palade on Ambystoma muscle were confirmed in the sartorius. In addition, the transverse tubules were observed to be continuous across the width of the fiber, a set of flat intermediate cisternae was seen to connect the terminal cisternae to the longitudinal tubules in the A band, and the continuous reticulum collar at the center of the A band was found to be perforated by circular and elongated pores (the fenestrated collar). The transverse tubules have a volume about 0.3 per cent of the fiber volume, and a surface area about 7 times the outer cylindrical surface area for a fiber 100 micro in diameter. The terminal cisternae, the intermediate cisternae, and the longitudinal tubules together with the fenestrated collar each have a volume of 4 to 5 per cent of the fiber volume and a surface area 40 to 50 times the outer surface area of a fiber 100 micro in diameter. Some evidence for continuity of the transverse tubules with the fiber surface is presented, but this is thought to be not so convincing as evidence presented by others. The results are discussed in terms of a possible mechanism for a role of the transverse tubules and sarcoplasmic reticulum in excitation-contraction coupling, as suggested by their morphology and a variety of physiological studies. In this scheme, the transverse tubules are thought to be electrically coupled to the terminal cisternae, so that depolarization of the fiber surface spreads inward along the transverse tubules and to the terminal cisternae, initiating the release of a contraction-activating substance.
622 citations