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PubMed1990

The effects of aldehydes on the inward rectifier of skeletal muscle fibres from the toad Bufo marinus.

A comparative study of the effects of formaldehyde and glutaraldehyde on the inward rectification in skeletal muscle fibre of the toad Bufo marinus was made using two different techniques. In whole sartorius muscle fibres formaldehyde produced a transient increase in conductance, followed by a decrease while glutaraldehyde only produced the second effect, with a rather fast time course. In cut end single fibres both aldehydes produced only a reduction of conductance and eventually the abolition of the inward rectification. The blocking effect of glutaraldehyde was concentration and voltage dependent while formaldehyde only produced a concentration dependent blocking. The elimination of inward rectifying currents by aldehydes allowed an estimation of the time constants of activation by a subtraction method.

Animals↗

PubMed1982

Pumiliotoxin-C and synthetic analogues. A new class of nicotinic antagonists.

Pumiliotoxin-C (PTX-CI), a cis-decahydroquinoline alkaloid, and synthetic analogues inhibited directly and indirectly elicited twitches in rat phrenic nerve diaphragm preparations. At 40 microM the most potent analogue, 2,5-di-n-propyl-cis-decahydroquinoline (PTX-CII), reversibly blocked the indirectly elicited twitch and markedly depressed the amplitude of end-plate potentials and miniature end-plate potentials without affecting muscle membrane potential. PTX-CII did not depress the directly elicited twitch in diaphragm muscle treated with alpha-bungarotoxin. At 40 microM, the compound blocked extrajunctional acetylcholine (ACh) sensitivity in chronically denervated soleus muscle of the rat. The peak amplitude of the end-plate current in frog sartorius muscle was depressed by PTX-CII at both positive and negative membrane potentials. Nonlinearity in the current-voltage relationship appeared at membrane potentials greater than -100 mV with concentrations of PTX-CII from 2.5 to 7.5 microM. The decay phase of the end-plate current was shifted to lower values such that it decayed faster at all membrane potentials but retained voltage sensitivity. PTX-CII, the most potent of the analogues, inhibited binding of [3H]perhydrohistrionicotoxin to Torpedo electroplax membranes with an IC50 value of 1.5 microM in the absence of carbamylcholine (Carb) and 0.1 microM in the presence of Carb. The other analogues were less potent, but all showed higher affinities in the presence of Carb. PTX-CI, PTX-CII, and PTX-CIII had no effect on the binding of [125I]alpha-bungarotoxin to ACh receptors in Torpedo membranes while slightly enhancing the binding of [3H]ACh, whereas PTX-CIV gave slight inhibition of both receptor ligands at concentrations greater than 10 microM. The pumiliotoxin-C class of alkaloids therefore appears to block neuromuscular transmission primarily via interactions with sites associated with the ion channel controlled by the ACh receptor.

Acetylcholinesterase↗

PubMed1993

Phosphate metabolism during muscular contraction in starved frogs (Rana catesbeiana).

1. The muscle tension and the state of high-energy phosphate metabolism during contraction of the sartorius muscle in frogs (Rana catesbeiana) starved for 1-5 months was studied by in vivo 31P-NMR spectrometry. 2. Muscle tension began to decrease after 2-month starvation compared with the control group and decreased to about one-third of the control value after a 5-month starvation. 3. Muscle contraction induced by electrical stimulation or the use of anaerobic perfusion fluid did not decrease the concentration of creatine phosphate (PCr) or beta-ATP, and only negligibly changed the PCr/Pi ratio from starvation. 4. These results suggest a decrease in creatine kinase activity in the muscle of starved frogs.

Animals↗

PubMed1984

Vanadate stimulates the pumped movements of Na in skeletal muscle.

We have measured the effects of concentrations of vanadate ranging between 0.01 and 10 mM on the 22Na efflux of frog sartorius muscles. The addition of vanadate had no effects when concentrations lower than 0.5 mM were used; higher concentrations increased Na efflux. The increase was abolished by the addition of ouabain (10(-5)M). In muscles pretreated with ouabain vanadate did not modify Na efflux. The stimulatory effects of vanadate on Na efflux were also observed in Na-free solutions indicating that the efflux of vanadate was not caused mainly either by an increase in the exchange of Na for Na or by an increase in Na entry into the muscle. We also examined the effects of vanadate on muscles immersed in solutions containing 20 mM K+; both vanadate and increased K+ produced stimulations of Na efflux that were additive. Similarly when the effects of vanadate and insulin were measured on the Na efflux of the same muscle, additive effects were found. As the ouabain-sensitive Na efflux in frog muscle is generally agreed to be due to the activity of the Na-K ATPase, our findings suggest that the net effect of vanadate in intact muscle cells is an increase in the activity of the Na pump. Since vanadate affects many enzymes it is quite possible that the stimulatory action is not due to a direct effect on the Na-K ATPase but may be mediated through an intermediary step.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1983

Stereospecificity of an opiate action on the excitable membrane of frog skeletal muscle fibres.

Intracellular microelectrode studies were conducted to examine the actions of the optical isomers levorphanol and dextrorphan on the isolated frog sartorius muscle preparation. Both isomers, in bath concentrations which had minimal effect on action potential amplitude and gNa (3 X 10(-5) M), caused a large depression of gK. High bath concentrations (3 X 10(-4) M) of both drugs caused an initial depression of sodium and potassium conductance processes, gNa and gK, respectively, however only levorphanol, but not dextrorphan, could produce a specific, late-occurring depression of gNa that could be blocked by the opiate antagonists, nalorphine or naloxone. These findings indicate that the late-occurring depressant effect on gNa is stereospecific and reinforce previous findings which demonstrated that opiate drugs interact with opiate receptors associated with the sodium channels of the frog muscle fibre membranes.

Action Potentials↗

PubMed1981

Millisecond time-resolved changes in x-ray reflections from contracting muscle during rapid mechanical transients, recorded using synchrotron radiation.

Low-angle x-ray diffraction diagrams have been recorded from frog sartorius muscles by using synchrotron radiation as a high-intensity x-ray source. This has enabled changes in some of the principal reflections of interest to be followed with a time resolution of 1 ms, during small but very rapid length changes imposed on a contracting muscle. The 143-A meridional reflection, which is believed to arise from a repeating pattern of myosin cross-bridges along the length of the muscle, shows large changes in intensity in these circumstances. During both rapid releases and rapid stretches, by amounts that produce a translation of actin and myosin filaments past each other by about 100 A and that are completed in about a millisecond (i.e., before significant cross-bridge detachment would be expected), an almost synchronous decrease in 143-A intensity occurs, by 50% or more. This is followed, in the case of quick releases, by a rapid partial recovery of intensity lasting 5--6 ms (which may represent cross-bridge release and reattachment) and then by a more gradual return to the normal isometric value. Quick stretches show only the slower return of intensity. Immediately after the length change, the initial drop in 143-A intensity can be reversed if the release (or stretch) is reversed. These changes provide evidence of a more direct kind than has hitherto been available that the active sliding of actin filaments past myosin filaments during contraction is produced by longitudinal movement of attached cross-bridges.

Actins↗

PubMed1980

Rubidium block and rubidium permeability of the inward rectifier of frog skeletal muscle fibres.

1. A three-electrode voltage clamp method was used to investigate the Rb block of inward rectification in frog sartorius muscle fibres. 2. In a solution containing 80 mM-K+, the potassium conductance increased with increasing hyperpolarization to 3.18 +/- 0.11 mS. cm-2 (n = 17) when (V - VK) was -150 mV. In the presence of Rb+, the conductance increased, fell and increased again with increasing hyperpolarization, i.e. the Rb block was first increased and then reduced by increasing hyperpolarization. Increasing [Rb]o increased the block at all voltages. 3. In a solution containing 80 mM-Rb+ (zero K+) inward currents were recorded when the membrane was hyperpolarized beyond about -60 mV. These currents, which were < 10% the amplitude of those in 80 mM-K solution, were blocked by tetraethylammonium ions. 4. Experiments were carried out in solutions either where both [K]o and [K]i were increased, or where [K]o only was increased. The form of the relation between K conductance and membrane potential appeared to depend on [K]o. The magnitude of the conductance appeared to depend on [K]o and on [K]i. 5. So far as the block by Rb+ is concerned, increasing [K]o appeared to enhance the release of Rb block under large hyperpolarizations. Increasing [K]o and [K]i reduced the Rb block at all membrane potentials. 6. The results of experiments in the presence of Rb+ and Cs+ suggest that these two ions do not compete with each other for a site at which they block inward rectification. Rather, over a range of membrane potentials from -25 to -65 mV, the presence of Cs+ enhances the Rb block and vice versa. 7. Single dissected muscle fibres (from semitendinosus) were used to measure sarcoplasmic resistivity in 80 mM-K solution and 160 mM-K (hyperosmotic) solution. The measured values were 163.2 +/- 11.7 omega x cm and 136.1 +/- 16.0 omega x cm, respectively (n = 7). 8. A semi-empirical model is presented, supposing that Rb interacts with a site in the membrane to produce its blocking effect, but is able to move on through into the sarcoplasm. Internal K+ is supposed to reduce the affinity of the site for Rb+; external K+ is able to enhance the moving on of Rb+ into the sarcoplasm. 9. The implications of our experiments for the nature of the permeability mechanism inward rectification are discussed.

Animals↗

PubMed1981

High-energy phosphate metabolism and energy liberation associated with rapid shortening in frog skeletal muscle.

1. High-energy phosphate metabolism and energy liberated as heat and work were measured in 3 sec tetani of frog sartorius muscles at 0 degrees C.2. Three contraction periods were studied: (a) shortening at near-maximum velocity for 0.3 sec from sarcomere length 2.6 to 1.8 mum, beginning after 2 sec of isometric stimulation, (b) the 0.7 sec isometric period immediately following such rapid shortening, (c) the period from 2 to 3 sec in an isometric tetanus at sarcomere length 1.8 mum.3. There were no significant changes in levels of ATP, ADP or AMP in any contraction period. The observed changes in inorganic phosphate and creatine levels indicated that the only significant reaction occurring was phosphocreatine splitting.4. The mean rate of high-energy phosphate splitting during rapid shortening, 0.48 +/- 0.24 mumole/g.sec (mean +/- s.e. of mean, n = 29; ;g' refers to blotted muscle weight), was not significantly different from that in the 1 sec period in the isometric tetanus, 0.32 +/- 0.11 mumole/g.sec (n = 17). The mean rate in the post-shortening period, 0.71 +/- 0.10 mumole/g.sec (n = 22), was greater than that in the 1 sec period in the isometric tetanus, and this difference is significant (P < 0.02, t test).5. A large quantity of heat plus work was produced during the rapid shortening period, but less than half of this could be accounted for by simultaneous chemical reactions. The unexplained enthalpy production was 6.5 +/- 2.6 mJ/g (mean +/- s.e. of mean). No significant unexplained enthalpy was produced in the 1 sec period in the isometric tetanus.6. In the post-shortening period the observed enthalpy was less, by 6.2 +/- 2.6 mJ/g, than that expected from the simultaneous chemical reactions.7. The results are interpreted in terms of an exothermic shift in the population of cross-bridge states during rapid shortening. It is suggested that a relatively slow subsequent step prevents many of these cross-bridges from completing the cycle and splitting ATP until after the end of shortening.

Adenosine Triphosphate↗

PubMed1990

Local oxygen supply and blood flow regulation in contracting muscle in dogs and rabbits.

1. Multiwire surface electrodes were used to measure local hydrogen clearance curves and tissues PO2 in the sartorius muscle in dogs under resting conditions and during stimulation of the muscle at 1, 2, 4, 8 and 20 Hz via the femoral nerve. Tissue oxygen supply was assessed by means of PO2 histograms; evaluation of the initial slopes of the hydrogen clearance curves enabled the measurement of capillary blood flow. 2. In a further model, the analysis of hydrogen clearance curves measured in the femoral vein using intravascular needle electrodes in rabbits enabled the distribution of blood flow to be evaluated both under resting conditions and during direct 2 Hz stimulation of the vastus medialis muscle. 3. Increased oxygen consumption, induced by stimulation, caused increases in capillary flow which were not necessarily accompanied by augmentation of femoral artery flow. 4. PO2 histograms provided no evidence of cellular anoxia even at the maximum level of oxygen consumption. 5. A two-compartment distribution of flow was measured under resting conditions, whereas only one compartment could be resolved during 2 Hz stimulation of the vastus medialis muscle in the rabbit experiments. A clear redistribution of flow was observed in the absence of any increase in total flow. 6. A model for oxygen-dependent regulation of capillary blood flow involving high-flow and normal-flow compartments is proposed.

Anesthesia, Intravenous↗

PubMed1979

Glyconeogenesis from lactate in frog striated muscle.

The rates of 14carbon incorporation into CO2 and glycogen from [U-14C]-lactate and [1-14C]acetate in frog sartorius muscles were compared. The rates of incorporation into CO2 were similar, while the rate of incorporation of lactate into glycogen was more than 200-fold larger than that of acetate incorporation. It is concluded that the pathway of lactate incorporation into glycogen does not involve Krebs cycle intermediates and is extramitochondrial. To test the possibility that the pathway of lactate incorporation involves net reversal of a pyruvate kinase, the changes in phosphoenolpyruvate and pyruvate concentrations during stimulation of lactate incorporation into glycogen were measured. There wer none. The mass action ratio of pyruvate kinase was calculated. This value was two orders of magnitude from the equilibrium constant and it was concluded that reversal of pyruvate kinase was a very unlikely pathway. To test the possibility that a pathway involving the oxaloacetate-to-phosphoenolpyruvate step was involved the muscles were treated with 3-mercaptopicolinate, an inhibitor of phosphoenolpyruvate carboxykinase. The treatment resulted in decreased incorporation of lactate into glycogen.

Acetates↗

PubMed1976

Lactate and contractile force in frog muscle during development of fatigue and recovery.

The relationship between lactic acid concentration and twitch tension was reevaluated in electrically stimulated frog sartorius muscle. In muscles stimulated under anaerobic conditions at a rate of 30 stimuli/MIN CONTRACTILE FORCE DECREASED TO 36% OF THE INITIAL VALUE IN 15 MIN, Concomitantly lactate increased from 3.3 to 18.7 mumol/g of muscle. The correlaiton between the increase in lactate and the decrease in contractile force was significant (r = -0.99, P less than 0.000001). Recovery occurred in two phases. A rapid increase in contractile force, which represented 20% of the total recovery, took place during the first 15 s and occurred concomitantly with an increase in ATP from 3.9 to 4.6 mumol/g. Lactate concentration did not change significantly during this period. The second phase of recovery of contractile force was complete in 50 min. Lactate concentration and contractile force were significatly correlated during recovery (r = -0;92, P less than 0.00001). However, recovery of contractile force lagged behind the decrease in lactate; a given concentration of muscle lactate was associated with a higher contractile force early during development of fatigue than late during recovery.

Adenosine Triphosphate↗

PubMed1985

[Effect of blockade of axonal transport on endplate currents of muscle fibers in the frog].

The evoked and spontaneous end-plate currents (EPC) were measured in normal voltage-clamped frog sartorius muscle fibres and after colchicine blockade of axoplasmic transport in the corresponding nerve. Colchicine application decreased the growth rate and increased the decay time constant of EPC without changes in the average amplitude of EPC and miniature EPC, quantum content of EPC and frequency of miniature EPC. The increase in EPC time constant was combined with an increase in the sensitivity of the time constant to holding potential, but there were no changes in the amplitude of EPC as a function of holding potential in the reversal potential of EPC. It is suggested that the increase in the EPC decay time constant was due to the appearance of a new population of receptor--ion channel complexes with opening time similar to that described for extrajunctional receptors after denervation. The neurotrophic control of chemosensitive muscle fibre membrane depended on the axoplasmic flow.

Animals↗

PubMedDate not supplied

[Effect of muscle storage and stimulation on the intensity of Z-reflection of its equatorial x-ray pattern].

The maintenance of frog sartorius muscle in Ringer solution over a long period of time (24 h or more) increases Z-reflection intensity of equatorial X-ray pattern relative to that from fresh (1-2 h) dissecting muscle at sarcomere length more than 2.3 m. Long stimulation deteriorates the equatorial X-ray pattern from muscle. Then with the course of time Z-reflection intensity increases several times, Z and reflections merge, and rigor equatorial X-ray pattern is registered.

Animals↗

PubMed1978

[Polyaxonal innervation of frog phasic muscle fibers].

A cholinesterase activity was revealed in two to four regions of each of the 120 fibers isolated from the frog sartorius muscle, which suggests polysynaptic character of innervation of the phasic muscle fibers. Electrophysiological methods showed the participation of different axons in formation of synapses with a single fiber, as well as the existence of synapses with the correlation between EPP's amplitude and strength of stimulation. The data obtained show such EPPs to be connected with polyaxonal innervation of one region of a muscle fiber.

Acetylcholinesterase↗

PubMed1981

The overall morphology of neuromuscular junctions as revealed by scanning electron microscopy.

Skeletal neuromuscular junctions (NMJs) of vertebrates were examined by scanning electron microscopy after removal of connective tissue components by HCl hydrolysis. In addition to the surface texture of NMJs, the subsynaptic organization of the sarcolemma was visualized in specimens in which nerve endings were detached from the muscle surface. A remarkable morphological variability between animal species was observed. The NMJs in the frog sartorius muscle consisted of longitudinal ribbon-like endings which fitted into a shallow synaptic gutter containing highly ordered cross-bands of junctional folds. The NMJs of the posterior latissimus dorsi muscle of the zebra finch were characterized by varicose swellings of the nerve endings which fitted into a round pit of the sarcolemma. NMJs in the sternothyroid muscle of the Chinese hamster consisted of thin ramified endings which were confined to an oval area on the muscle surface. The labyrinthine synaptic groove contained well-developed junctional folds without preferential spatial arrangement. The procedure used for the present study illustrates in great detail the terminal arborization of the motor nerve ending and the surface features of the subsynaptic sarcolemma. It may also allow quantitative study of the synaptic morphology of NMJs.

Animals↗

PubMed1981

Comparison of the action of La3+ and Ca2+ on contraction threshold and other membrane parameters of frog skeletal muscle.

The influence of La3+ on contraction threshold, on membrane input resistance, and on action potential parameters was investigated in fibers of the sartorius muscle of the frog, and it was compared to that of Ca2+. The dependence of the contraction threshold on [La3+]0 in the presence of 0.5mM Ca3+ gave a sigmoid relationship between 0.1 and 5 mM La3+ with a shift of 23 to 34 mV to less negative potentials following a 10-fold increase of [La3+]0. The membrane input resistance was increased to various degrees in La-containing solutions, the increase being irreversible. The threshold of action potential generation was shifted to less negative potentials by 28 mV, and the duration at half-maximal amplitude was tripled by 0.5 mM La3+. In comparison a 10-fold increase of [Ca2+]0 in the range of 0.5 to 50 mM shifted the contraction threshold by 15 mV to less negative potentials. 17 mM Ca3+, a concentration having the same effect on contraction threshold as 0.5 mM La3+, increased membrane input resistance reversibly, shifted the action potential threshold by 16 mV to less negative potentials, and had only minor effects on action potential duration. Conduction was never blocked by Ca3+ as it was with 1 mM La3+. In the theoretical treatment, it is shown that the influence of Ca3+ on contraction threshold, but not that of La3+, may be accounted for by its screening and binding to negative surface charges according to the Gouy-Chapman theory of the diffuse double layer. To describe the action of La3+ on the contraction threshold an additional interaction of La3+ with neutral but amphoteric sites was considered.

Action Potentials↗

PubMed1990

Studies of the diffuse x-ray scattering from contracting frog skeletal muscles.

Using x-rays from synchrotron radiation, we studied diffuse scattering, sometimes together with the myosin layer lines. With an area detector, sartorius muscles and a time resolution of 150 ms, earlier results from semitendinosus muscles contracting isometrically at 6 degrees C (Lowy, J., and F. R. Poulsen. 1987. J. Mol. Biol. 194:595-600) were confirmed and extended. Evidence from intensity changes both in the diffuse scattering and in the myosin layer lines showed that the majority of the heads become disordered at peak tetanic tension. With a linear detector and a time resolution of 5 ms, it was found that during tension rise the intensity increase of the diffuse scattering (which amounted maximally to 12% recorded near the meridian) runs approximately 20 ms ahead of the mechanical change, comparing half-completion times. This suggests that an appreciable number of heads change orientation before peak tension is reached. In quick release experiments the diffuse scattering intensity showed very little change. Recorded near the meridian during rapid shortening, however, it decreased progressively with a half-time of approximately 40 ms. This change amounted to approximately 35% of that observed during the initial tension rise. We interpret this to indicate that during rapid shortening a certain number of heads assume an orientation characteristic of the relaxed state. Viewed in the context of the behavior of the first myosin layer line and the (1, 1) equatorial reflection in similar experiments (Huxley, H. E., M. Kress, A. R. Faruqi, and R. M. Simmons. 1988. Molecular Mechanism of Muscle Contraction), the present results provide further support for the view that the diffuse scattering is mostly due to disordered myosin heads; whilst ordered heads produce the myosin layer lines (Poulsen, F. R., and J. Lowy.1983. Nature lLond.l. 303:146-152).

Animals↗

PubMed1965

The variation of muscle oxygen consumption with velocity of shortening.

Total oxygen consumption following contraction at controlled shortening velocities was determined for ten twitches of frog sartorius muscle. Under the conditions of this investigation maximum oxygen consumption occurred at approximately 30 mm/sec. shortening velocity and decreased for larger and smaller velocities. Mechanical efficiency was calculated and found to show a maximum at approximately 20 mm/sec. shortening velocity. The relationship between this study and a similar investigation in which heat changes are measured as a function of shortening velocity is discussed.

Animals↗