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R S Wilkinson

Publications and source records attributed to R S Wilkinson.

11 recordsLinked to original sources
PubMed1992

Regulation of single quantal efficacy at the snake neuromuscular junction.

1. Postsynaptic responses to spontaneous quantal transmitter release have been compared among neuromuscular junctions in a thin snake muscle. For each junction the type, diameter, and input conductance, G(in) of the postsynaptic muscle fibre were determined. Particularly among fibres of a given type, G(in) was directly correlated with fibre diameter. 2. Miniature endplate potentials (MEPPs) were recorded intracellularly near endplates visualized with Nomarski optics. Mean MEPP amplitude decreased with increasing G(in) among fibres in one muscle. However, the dependence of mean amplitude upon G(in) was not ohmic, as would be expected if the underlying single quantal currents (miniature endplate currents, MEPCs) were of similar amplitude at all junctions. Instead, the relation between MEPPs and G(in) suggested that mean MEPC amplitudes, calculated as mean MEPP amplitude x G(in), increased with increasing G(in). 3. MEPCs were recorded directly using the two-microelectrode voltage clamp technique. Mean MEPC amplitudes depended systematically on G(in), again such that MEPCs were on average larger in fibres with higher G(in). 4. MEPCs were recorded extracellularly from small regions of endplates (underlying a few nerve terminal boutons). Amplitudes of MEPCs depended on G(in) or fibre diameter in the same manner as amplitudes of MEPCs recorded by intracellular voltage clamp. 5. When the anticholinesterase agent neostigmine was added to the bath, amplitude and duration of MEPPs, MEPCs, and extracellular MEPCs increased. However, the systematic dependence of mean MEPC amplitude on G(in) or fibre diameter remained. 6. Evoked subthreshold endplate potentials (EPPs) were recorded under conditions of low extracellular Ca2+. Endplate currents (EPP amplitudes x G(in)) were systematically larger in fibres with larger G(in), indicating regulation of evoked synaptic current in the muscle. The regulation was found to be due to a combination of increased quantal content and larger single quantal currents in larger (higher G(in)) fibres. 7. Synaptic size, assessed either by area of cholinesterase staining or number of terminal boutons, increased with increasing fibre diameter. Assuming that quantal content is proportional to synaptic size, this relation was sufficient to account for the observed increase in quantal content with increasing G(in) among fibres in the muscle, but was not alone sufficient to account for the observed regulation of evoked current. 8. It is concluded that the efficacy of individual transmitter quanta released at the snake neuromuscular junction is regulated such that large muscle fibres receive larger single quantal currents. Regulation of single quantal current contributes substantially to overall regulation of synaptic strength in the muscle.

Animals↗
PubMed1991

Metabolic and contractile uniformity of isolated motor unit fibres of snake muscle.

1. Motor units in the thin transversus abdominis muscle of the garter snake were identified and physiologically characterized in the living state. Motor unit fibres, and fibres chosen randomly to serve as controls, were subsequently excised and subjected to biochemical analyses. 2. The metabolic capacity of fibres was assessed by measuring activities of three enzymes, each representing a different metabolic pathway. The microchemical enzyme assays were performed using enzyme extraction preparations of whole single fibres. 3. Metabolic capacity ranged widely among the muscle's entire fibre population, even among fibres of the same type. In contrast, enzyme activities of twitch fibres belonging to individual motor units were, within analytical error, identical. 4. Twitch contraction times of individual fibres within one motor unit were similar, compared to a wide range of contraction times observed among fibres of the same type but belonging to different motor units. 5. When several motor units were studied in one muscle, a systematic relationship was observed among motor unit tension, enzymatic profile and contraction time. As motor unit tension increased, fibres exhibited greater capacities for glycolytic and high-energy phosphate metabolism, diminished capacity for oxidative metabolism, and faster twitch contraction times. 6. Given the great diversity of metabolic and contractile properties exhibited within the fibre population, the uniformity of such properties within motor units indicates that neural influence dominates over other extrinsic factors present in the microenvironment of the muscle fibres.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed1991

Metabolic capacity and myosin expression in single muscle fibres of the garter snake.

1. The transversus abdominis muscle of the garter snake contains fibres of three types: tonic (T), slower twitch (S) and faster twitch (F). Fibre types can be determined by anatomical criteria in living preparations. Individual fibres identified as T, S or F were excised from the muscle and subdivided for two types of biochemical examination. Enzymes of energy metabolism were assayed using quantitative microfluorometric methods. Myosin heavy chain composition was determined by gel electrophoresis. In separate experiments, twitch time-to-peaks of F and S fibres were measured to assess the range of contraction times present within the muscle's twitch fibre population. 2. Metabolic subgroups of fibres were delineated by the relative activities of adenylokinase (AK), lactate dehydrogenase (LDH) and beta-hydroxyacyl-CoA-dehydrogenase (beta OAC). The metabolic subgroups corresponded to the anatomical fibre types. Type F fibres had high levels of enzymes associated with glycolytic (LDH) and high-energy phosphate (AK) metabolism. Type T fibres had high levels of the oxidative enzyme beta OAC. Type S fibres had both types of enzyme activity in intermediate and variable amounts. 3. Three myosin heavy chain isoforms were present in the muscle. Type F and type T fibres each expressed a single isoform, denoted F and T respectively. Type S fibres expressed significant quantities of two isoforms: an isoform unique to this fibre type (denoted S) and the F isoform. 4. Electrophoretic mobility and antibody reactivity of the F myosin heavy chain isoform resembled that of mammalian fast-twitch myosin. By the same criteria, the T isoform resembled mammalian slow-twitch myosin. The S isoform exhibited intermediate characteristics: its antibody reactivity was similar to mammalian fast-twitch myosin, but its electrophoretic mobility was that of mammalian slow-twitch myosin. 5. Based on whole-muscle analysis, two myosin alkali light chains, denoted ALC1 and ALC2, and one myosin regulatory light chain were present. Gel patterns suggested that ALC1 and ALC2 exist as both homodimers and heterodimers. 6. The population of type S fibres within a given muscle exhibited a much wider range of twitch contraction times than did the population of type F fibres. Diversity of contractile properties among type S fibres may result, in part, from differential co-expression of two myosin heavy chain isoforms, together with highly variable ratios of enzymes from two major metabolic pathways. 7. The clear biochemical distinction among fibre types indicates that each type possesses a unique and limited range of physiological properties.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

The effect of reinnervation on the distribution of muscle fibre types in the tibialis anterior muscle of the mouse.

The distribution of fibre types in the tibialis anterior (TA) muscle of adult mice was examined by means of an immunohistochemical approach, using monoclonal antibodies that recognize different myosin heavy chain isoforms. As has been reported previously, the superficial portion of TA contains almost exclusively type IIB fibres and is almost entirely glycolytic in nature. Following section of the lateral popliteal nerve and rotation of the proximal stump to prevent rematching, it was found that the original pattern was virtually restored within 2 months. One possible explanation for this observation is that the activity pattern of peripheral and deep muscle fibres differs and that this aids in specification of muscle fibre type. Alternatively, the muscle fibres of the superficial portion of TA may be inherently resistant to an alteration of their phenotype with regard to expression of myosin heavy chain.

Adenosine Triphosphatases↗
PubMed1989

High-resolution imaging of synaptic structure with a simple confocal microscope.

We describe a straightforward modification to a conventional fluorescence microscope that permits confocal fluorescence imaging of living and fixed biological material. The modified microscope has been used to examine the morphology of presynaptic terminals at the snake neuromuscular junction. Among our observations is the presence of two discrete compartments within each terminal bouton.

Animals↗
PubMed1985

Regular alternation of fiber types in the transversus abdominis muscle of the garter snake.

The snake transversus abdominis muscle is an extremely simple segmentally repeating muscle containing 80 to 100 muscle fibers in a single-fiber-thick sheet. This muscle exhibits a striking pattern of muscle fiber types: twitch fibers alternate with tonic fibers and, among the twitch fibers, slower and faster contracting subtypes also alternate. Thus, in many regions of the muscle the pattern of fiber types is: faster twitch, tonic, slower twitch, tonic, faster twitch, tonic, and so on. The existence of a spatial pattern of fiber types, perhaps discernible in this muscle because of the muscle's extreme geometrical simplicity, provides good evidence for an intrinsic component to muscle fiber differentiation.

Action Potentials↗
PubMed1979

Mixed lymphocyte reactions in serum-free medium.

Tissue culture medium supplemented with bovine serum albumin (BSA), rather than human serum, was found capable of supporting the proliferative response of cryopreserved human lymphocytes to allogeneic cells in a one-way mixed lymphocyte culture. Although total tritiated thymidine uptake was lower in BSA supplemented cultures than in serum supplemented cultures, stimulation indices were equivalent. Using 0.5% (w/v) BSA, peak tritiated thymidine uptake was observed on day 5 using 2 X 10(5) responders and 4X 10(5) stimulators per microtitre well. Calf skin gelatin was also found capable of substituting for serum, but gave lower levels of proliferation. Use of BSA offers distinct advantages for investigating the proliferative responses of human lymphocytes in serum-free media.

B-Lymphocytes↗
PubMed1978

Ionic basis of the receptor potential in primary endings of mammalian muscle spindles.

The effect of changing the ionic composition of bathing fluid on the receptor potential of primary endings has been examined in isolated mammalian spindles whose capsule was removed in the sensory region. After impulse activity is blocked by tetrodotoxin, ramp-and-hold stretch evokes a characteristic pattern of potential change consisting of a greater dynamic depolarization during the ramp phase and a smaller static depolarization during the hold phase. After a high-velocity ramp there is a transient post-dynamic undershoot to below the static level. On release from hold stretch, the potential shows a postrelease undershoot relative to base line. The depolarization produced by stretch is rapidly decreased by the removal of Na+ and Ca2+. Addition of normal Ca2+ partly restores the response. Stretch appears to increase the conductance to Na+ and Ca2+ in the sensory terminals. The postdynamic undershoot is diminished by raising external K+ and blocked by tetraethylammonium (TEA). It apparently results from a voltage-dependent potassium conductance. The postrelease undershoot is decreased by raising external K+, but is not blocked by TEA. It is presumably caused by a relative increase in potassium conductance on release. Substitution of isethionate for Cl- or the addition of ouabain does not alter the postdynamic and postrelease undershoots.

Action Potentials↗
PubMed1977

Responses of isolated Golgi tendon organs of the cat.

1. The responses to stretch have been studied in living, isolated Golgi tendon organs (GTOs) from tail muscles of cat. Experiments were performed in vitro and consisted of subjecting single GTOs to controlled ramp-and-hold stretch while recording the response from their sensory axons raised in oil. 2. The threshold force required for sustained afferent discharge was measured directly, and found to be between 8 and 170 dynes at 24 degrees C for nine GTOs tested. Beyond threshold, the discharge frequency is approximately proportional to applied static tension over a wide range. Sensitivy to tension varies among different GTOs and appears to be inversely correlated with mechanical stiffness. 3. With impulse activity blocked by tetrodotoxin, graded receptor potentials could be recorded whose amplitude varied in proportion to applied static tension. All GTOs examined showed in addition a dynamic response, which became larger with increasing velocity of ramp stretch. This dynamic sensitivity appears in the receptor potential and is then augmentd by an apparent accommodative process at the impulse initiating site. 4. Based on the above findings, possible mechanical models are discussed for the sensory transduction mechanism.

Action Potentials↗