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PubMed1988

Ion permeation through single ACh-activated channels in denervated adult toad sartorius skeletal muscle fibres: effect of temperature.

The gigaohm seal technique was used to study the effects of temperature on ion permeation through acetylcholine-activated channels. This was done in cell-attached patches of the extrajunctional membrane of chronically-denervated, enzyme-treated cells from sartorius muscle of the toad Bufo marinus. The predominant extracellular cation in the pipette solution was Na+. Single channel current-voltage curves were measured at different temperatures and electrodiffusion and three-site-four-barrier rate theory models were used to characterize ion permeation through the channels and determine the effects of temperature on permeation parameters. The fitting of the experimental data to these models suggested the presence of at least three and probably more ion-selective sites within the channel. The most frequently occurring channel type (greater than 95% of channel openings) had a chord conductance of 25 pS at 11 degrees C and -70 mV and was classified as 'extrajunctional'. The single channel conductance of this channel had a low temperature-dependence (Q10 approximately equal to 1.3). The apparent activation enthalpy, Ea, for the conductance between 11 degrees C and 20 degrees C, did not appear to be significantly voltage-sensitive and had a value of about 17 +/- 2 kJ . mol-1 at a voltage of -70 mV. The Arrhenius plot of conductance appeared linear between 11 and 20 degrees C at all potentials examined. The data was consistent with a break in the slope of the Arrhenius plot at temperatures between 5 and 11 degrees C at all potentials examined, suggesting a possible phase transition of the membrane lipids. In contrast to the relative permeability, which was not very temperature sensitive, the relative binding constant was significantly affected by temperature. The relative Na/K binding constant sequence was: K5 degrees C greater than K20 degrees C greater than K15 degrees C much greater than K11 degrees C. In addition, the decrease in conductance observed at the most depolarized potentials was accentuated as the temperature was increased, suggesting a rate-limiting access step for ions from the intracellular solution into the channel.

Acetylcholine↗

PubMed1976

Effect of growth and plane of nutrition on apparent muscle fiber numbers in the pig.

A histological sampling technique was used to estimate the number of muscle fibers appearing in transverse sections of whole sartorius muscles. Transverse sections were taken at the midlength of sartorius muscles from pigs growing at a rate of approximately 0.7 kg/day. Between 56 and 168 days (postnatal), the apparent number of muscle fibers increased at an approximate rate of 300 fibers/day. No evidence of the formation of new muscle fibers by myoblast fusion or muscle fiber fission was detected by light microscopy. Animals placed on a maintenance diet which prevented growth for 28-day periods prior to slaughter showed a fixation or decrease in apparent numbers of muscle fibers. These data support the hypothesis that the arrangement of muscle fibers during growth of the porcine sartorius muscle is a dynamic rather than a fixed system.

Animal Nutritional Physiological Phenomena↗

PubMed1980

The effect of diazepam on tension and electrolyte distribution in frog muscle.

The effect of diazepam on sartorius muscles of the frog was evaluated. Resting tension in sartorius muscle was not affected by diazepam (5 x 10(-5)M) but twitch tension was increased and tetanus tension decreased. The kinetics of 45Ca efflux were altered by diazepam. The calcium content of the intermediate pool was increased by diazepam (5 x 10(-6)M). When the diazepam concentration was increased (5 x 10(-5)M), the time constant of the slow pool decreased and the 45Ca content of the intermediate pool increased further. It is suggested that diazepam interferes with the calcium sequestering system of the sarcoplasmic reticulum (slow pool) and causes an increase of the calcium content of the myofibrillar space (intermediate pool).

Animals↗

PubMed1970

The Meyerhof quotient and the synthesis of glycogen from lactate in frog and rabbit muscle.

1. The conversion of lactate into glycogen was demonstrated in frog sartorius muscle in oxygen. The rates and amounts are highest when lactate is added to the bathing medium and are dependent on lactate and CO(2) concentration, as well as pH. The glycogen content of a resting muscle can be doubled in 4h at 24 degrees C. 2. Sartorius muscle, recovering aerobically in liquid paraffin from a period of anoxia, converts preformed lactate into glycogen at a lower rate and in smaller amounts than when lactate is added in an aqueous medium. The lower rates are similar to those Meyerhof found under the same conditions, after correction for temperature; they can be attributed partly to low muscle pH and partly to the limited amounts of lactate present. 3. Rabbit psoas muscle also shows the ability to convert added lactate into glycogen under aerobic conditions. The rates are low and similar to those in frog sartorius muscle recovering from anoxia. 4. The present experiments yield a Meyerhof quotient of 6.2, compared with Meyerhof's value of 4-5. However, these values are not significantly different from one another. 5. It is suggested that the glycogen coefficient, i.e. mol of glycogen formed/mol of lactate disappearing, is a more reliable way of assessing the resynthetic mechanism than the original quotient, i.e. mol of lactate disappearing/mol of lactate oxidized. The found coefficient is 0.419+/-0.024.

Animals↗

PubMed2003

Phosphate metabolites in muscular contraction caused by magnetic stimulation.

The metabolism of high energy phosphates during muscular contraction due to direct electrical stimulation, indirect stimulation via nerve excitation, and magnetic stimulation was studied in isolated muscles (frog sartorius muscles) by (31)P nuclear magnetic resonance ((31)P-NMR). Twitch amplitudes elicited by each stimulus were measured alternatively at 3 mm displacement loading and 5 g weight. Both the creatine/inorganic phosphate (PCr/Pi) and pH changes were more marked in direct electrical stimulation than in magnetic stimulation. The muscular contraction caused by magnetic stimulation showed less fatigue than that caused by direct electrical muscular stimulation.

Animals↗

PubMedDate not supplied

The effects of sodium nitroprusside on mediator release and the functional properties of postsynaptic membranes in the neuromuscular synapse.

Experiments on neuromuscular preparations of frog skin- thoracic muscle and sartorius muscle, using extracellular recording and two-electrode clamping of the muscle fiber membrane potential, were used to study the effects of the nitric oxide donor sodium nitroprusside on endplate currents. At a concentration of 100 microM, sodium nitroprusside sharply decreased the amplitude and quantum composition of the endplate currents, and also decreased the miniature endplate current frequency. The amplitude-time characteristics of miniature endplate currents, the voltage-dependent amplitude, and the decay time constant of miniature endplate currents did not change as compared with controls. However, unlike the situation with other secretion inhibitors, the decrease in endplate current amplitude was not accompanied by increased facilitation in response to rhythmic stimulation or changes in postsynaptic potentiation in conditions of application of pairs of stimuli to muscles. The suppression of acetylcholine secretion was not seen with inactivated sodium nitroprusside solution. These results provide evidence that nitric oxide can be a powerful inhibitor of both spontaneous and evoked transmitter secretion in the neuromuscular synapse, and that this is accompanied by decreases in the efficiency of presynaptic forms of short-term plasticity, while the functional characteristics of the postsynaptic membrane remain unchanged.

Action Potentials↗

PubMed1995

Comparative contractile dynamics of calling and locomotor muscles in three hylid frogs.

Isometric twitch and tetanus parameters, force-velocity curves, maximum shortening velocity (Vmax) and percentage relaxation between stimuli (%R) across a range of stimulus frequencies were determined for a muscle used during call production (the tensor chordarum) and a locomotor muscle (the sartorius) for three species of hylid frogs, Hyla chrysoscelis, H. versicolor and H. cinerea. The call of H. chrysoscelis has a note repetition rate (NRR) approximately twice as fast as the call of H. versicolor (28.3, 42.5 and 56.8 notes s-1 for H. chrysoscelis and 14.8, 21.1 and 27.4 notes s-1 for H. versicolor at 15, 20 and 25 degrees C, respectively). Hyla cinerea calls at a very slow NRR (Approximately 3 notes s-1 at 25 degrees C). Hyla versicolor evolved from H. chrysoscelis via autopolyploidy, so the mating call of H. chrysoscelis is presumably the ancestral mating call of H. versicolor. For the tensor chordarum of H. chrysoscelis, H. versicolor and H. cinerea at 25 degrees C, mean twitch duration (19.2, 30.0 and 52.9 ms, respectively), maximum isometric tension (P0; 55.0, 94.4 and 180.5 kN m-2, respectively), tetanic half-relaxation time (17.2, 28.7 and 60.6 ms, respectively) and Vmax (4.7, 5.2 and 2.1 lengths s-1, respectively) differed significantly (P < 0.05) among all three species. The average time of tetanic contraction to half-P0 did not differ significantly between H. chrysoscelis (14.5 ms) and H. versicolor (15.8 ms) but was significantly longer for H. cinerea (52.6 ms). At 25 degrees C, Vmax differed significantly among the sartorius muscles of H. chrysoscelis, H. versicolor and H. cinerea (5.2, 7.0 and 9.8 lengths s-1, respectively) but mean twitch duration (29.5, 32.2 and 38.7 ms, respectively), P0 (252.2, 240.7 and 285.1 kN m-2, respectively) and tetanic half-relaxation time (56.3, 59.5 and 60.7 ms, respectively) did not differ significantly. The average time of contraction to half-P0 did not differ significantly between H. chrysoscelis (23.7 ms) and H. versicolor (22.9 ms) but was significantly shorter for H. cinerea (15.6 ms). The only consistent contractile differences found in this study between the calling muscle and locomotor muscle of H. chrysoscelis, H. versicolor and H. cinerea were that the calling muscles generated less tension and their force-velocity relationship was much more linear. These differences may be attributable to ultrastructural differences between calling and locomotor muscles.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1979

The physiology, pharmacology, and trophic effectiveness of synapses formed by autonomic preganglionic nerves on frog skeletal muscle.

1. Frog sartorius muscles, newly denervated and transplanted to the lymph sac of the back, are reinnervated by implanted cholinergic nerves (spinal somatic motor nerves or the preganglionic sympathetic splanchnic nerve), but not by nerves). 2. Foreign somatic motor nerves (s.m.n.s) form synapses that resemble normal sartorius neuromuscular junctions electrophysiologically. 3. Axons of the sympathetic preganglionic splanchnic nerve (s.p.n.) grow throughout the muscle, but only a small percentage of fibres form synapses. Most e.p.p.s are of low quantal content, generally subthreshold. Long onset latencies and multiple post-synaptic responses indicate that innervation is multiple, multi-terminal, and by unmyelinated axons. 4. Spontaneous miniature e.p.p.s at splanchnic junctions occur at an average rate under 0.1/sec. Their average amplitude and time course are about the same as for control muscles, but the variability of amplitudes is greater than for control muscles. 5. The amount of facilitation shown by s.p.n.-evoked e.p.p.s is the same as by s.m.n. e.p.p.s, but the time course is almost twice as long. 6. S.p.n.-reinnervated fibres show dramatic post-tetanic potentiation preceded by depression, following as few as 20--50 stimuli. 7. As judged by standard physiological and histochemical criteria, AChEsterase is absent at s.p.n. junctions. 8. The pharmacological responses of the s.p.n. junctions are similar to those of normal or foreign s.m.n. innervated neuromuscular junctions in their sensitivity to the cholinergic blocking agents D-tubocurarine and hexamethonium. 9 The s.p.n. is capable of restricting ACh sensitivity to the sites of nerve contacts, although this restriction occurs more slowly and less completely than with s.m.n. reinnervation. The loss of extrajunctional ACh sensitivity can be correlated with effectiveness of innervation; but significant restriction occurs even in s.p.n. reinnervated fibres that probably never contract to nerve stimulation.

Acetylcholine↗

PubMed1997

Anatomical course of the lateral femoral cutaneous nerve and its susceptibility to compression and injury.

The anatomy of the lateral femoral cutaneous nerve was investigated through dissection of 52 human anatomic specimens. The variability of its course and locations as it exists the pelvis is described and related to soft-tissue and bony landmarks. Five different types are identified: type A, posterior to the anterior superior iliac spine, across the iliac crest (4 percent); type B, anterior to the anterior superior iliac spine and superficial to the origin of the sartorius muscle but within the substance of the inguinal ligament (27 percent); type C, medial to the anterior superior iliac spine, ensheathed in the tendinous origin of the sartorius muscle (23 percent); type D, medial to the origin of the sartorius muscle located in an interval between the tendon of the sartorius muscle and thick fascia of the iliopsoas muscle deep to the inguinal ligament (26 percent); and type E, most medial and embedded in loose connective tissue, deep to the inguinal ligament, overlying the thin fascia of the iliopsoas muscle, and contributing the femoral branch of the genitofemoral nerve (20 percent). The results of this study suggest that the lateral femoral cutaneous nerve is most susceptible to mechanical trauma when the nerve is type A, B, or C.

Humans↗

PubMed1986

Labile heat and changes in rate of relaxation of frog muscles.

Observations were made of the labile heat and the progressive slowing of relaxation as a function of tetanus duration (Abbott effect) during isometric tetani of frog extensor longus digiti IV (e.l.d. iv) and sartorius muscle. Both the labile heat and slowing of relaxation are less marked in e.l.d. iv than in sartorius muscle. Both effects are depressed to the same extent in the second of two closely spaced tetani in sartorius muscle. The repriming of both effects follows the same time course in sartorius muscle. The hypothesis is discussed that both effects result from binding of calcium to parvalbumin during a tetanus, and that their repriming is due to the removal of calcium from parvalbumin by the sarcoplasmic reticulum.

Animals↗

PubMed1978

Increased cyclic GMP in the end-plate region of denervated frog muscle.

Denervated frog sartorius muscles showed an approximately 2--3 fold increase of cyclic GMP in their end-plate rich regions which did not appear up to 5 weeks after denervation in the normally end-plate-free pelvic region. No increase in cyclic AMP was seen in these preparations. The results suggest that the increase of cyclic GMP is related to processes specific to the region in which end plates are normally present.

Acetylcholine↗

PubMed1978

Neurotrophic control of RNA synthesis in amphibian striated muscle.

When frog sartorius muscles are incubated in culture for 3 days, a 98% increase in RNA synthesis by the muscles is seen. A portion of this increase (60--100% in different experiments) is prevented by the presence of a 2 cm distal nerve stump left attached to the muscle, or by the addition to the culture medium of nerve and spinal cord extracts from frog, rat or cow. The effect of the nerve stump and the extract is seen under conditions of neuro-muscular blockade by 50 mg/l gentamicin sulfate present in the culture medium. Autoradiography of 1 micrometer sections of muscles previously labeled with [3H]uridine shows that isotope incorporation is largely confined to muscle fibers rather than connective tissue. Muscles cultured for 3 days display normal action and resting potentials either in the presence or absence of extract. The results are discussed in terms of control of muscle RNA synthesis by substances present in motor neurons.

Action Potentials↗

PubMed1980

The effect of lanthanum ions on acetylcholine in frog muscle.

1. Frog sartorius muscles were treated with an irreversible cholinesterase inhibitor and then incubated in Ringer with 2 mM-LaCl3. The amounts of ACh in the tissue and medium were assayed by mass fragmentography, miniature end-plate potentials (min. e.p.p.s) were recorded and the end-plate was investigated by electron microscopy. 2. Addition of La3+ caused in normal, but not in denervated, muscles a discharge of both min. e.p.p.s and chemically detectable ACh. After 30 min both min. e.p.p.s and ACh release decreased. Between 4 and 5 hr after the addition of La3+ min. e.p.p.s had practically ceased and the rate of ACh release was almost back to that in the absence of La3+. 3. La3+ caused a 50% reduction in the ACh content of the tissue within the first 30 min; thereafter ACh gradually increased to 110% by 5 hr. At this time synaptic vesicles were practically absent in most terminals. The ACh was predominantly located in the end-plate regions of the muscles, before as well as after the incubation with La3+. ACh in end-plate free parts of the muscles was unchanged by La3+. 4. Hemicholinium-3 inhibited the synthesis of ACh in the muscles, but it had almost no influence on La3+-induced ACh release. 5. From these and other results, it is concluded that the ACh released by La3+ originates exclusively from the nerve terminals, that most likely this ACh is released via exocytosis from synaptic vesicles, and that the synthesis of ACh following the release of ACh takes place in the nerve terminals. The results further indicate that in freshly excised muscle the greater part (80-90%) of the ACh contained in the nerve terminals is located in the vesicles.

Acetylcholine↗

PubMed1982

Free and bound acetylcholine in frog muscle.

1. Frog sartorius muscles were divided into end-plate containing (e.p.) and end-plate-free (non-e.p.) segments or homogenized in Ringer solution at 0 degrees C in the presence or absence of added acetylcholinesterase from electric eel. ACh was extracted from the tissue or from the homogenates and measured by mass fragmentography. 2. The concentration of ACh in non-e.p. segments was about six times lower than that in e.p. segments. 3. Homogenization of muscles in Ringer caused the hydrolysis of a small fraction ('free-1') of total ACh; addition of extra acetylcholinesterase caused hydrolysis of another, greater, fraction ('free-2' ACh). The esterase-resistant ('bound') ACh was stable at 0 degrees C up to 15 min of incubation. 4. Denervation for 15 days, which caused the disappearance of the nerve terminals, did not influence ACh in non-e.p. segments, but reduced total and bound ACh by about 75%, and free-2 ACh by 90%. 5. Treatment with La3+ ions, which caused the disappearance of synaptic vesicles, did not influence total ACh, but reduced bound ACh by 75%, whereas free-1 and free-2 ACh were increased. 6. Electrical stimulation of the nerve at 5 sec-1 or incubation with 50 mM-KCl did not affect ACh in the non-e.p. segments, but reduced by roughly 60% total, bound, and free ACh. 7. It is concluded that about 75% of bound ACh derives from synaptic vesicles, corresponding to 11,000 molecules per vesicle, and 25% from non-neural ACh; that free-1 and free-2 ACh derive mainly from the nerve terminal cytoplasm, although they may be contaminated by vesicular ACh.

Acetylcholine↗

PubMed1971

Sodium and potassium activities in normal and "sodium-rich" frog skeletal muscle.

In frog sartorius muscles immersed for 2 hours at 26 degrees C in normal Ringer solution, the intrafiber potassium concentration, C(K)(in millimoles per liter), was 123 +/- 2 (mean value plus or minus standard error), and the potassium activity, a(K) (in millimoles per liter), was 90 +/- 1.0. The corresponding sodium concentration and activity were 20 +/- 1 and 6.5 +/- 0.4, respectively. After overnight immersion in K+-free Ringer solution the values were: CK, 97 +/- 2; aK, 81.5 +/- 1.6; CNa, 47 +/- 2; and aNa, 11.2 +/- 0.6. The changes in aK and aNa during storage were not consistent with an exchange between predominantly "free" fiber K+ and external Na+. These results suggest that the Na+ taken up during overnight immersion largely replaced adsorbed or sequestered K+ in the fibers.

Animals↗

PubMed1989

In vitro Na+,K+-ATPase (EC 3.6.1.3)-dependent respiration and protein synthesis in skeletal muscle of pigs fed at three dietary protein levels.

1. Eighteen pigs were offered diets containing 130, 170 or 210 g protein/kg with three barrows and three gifts per diet from 20 to 60 kg live weight. Oxygen consumption, Na+,K+-ATPase (EC 3.6.1.3)-dependent and -independent respiration and protein synthesis were measured in vitro in intercostal and sartorius muscle preparations from these pigs. 2. Increasing dietary protein concentration increased (P less than 0.01) daily gain and dissectible muscle in carcass. 3. O2 consumption and Na+,K+-ATPase-dependent respiration of the intercostal and sartorius muscles increased linearly (P less than 0.01) with increase in dietary protein concentration. The requirement for the support of the transport of Na+ and K+ across the cell membrane in these muscles, on average, accounted for 22-25% of the O2 consumption. 4. Synthesis rate (mg/g per d) of protein in the sartorius muscle increased (P less than 0.05) from 3.05 to 5.07 and increased (P less than 0.1) from 2.57 to 4.06 in the intercostal muscle as dietary protein increased from 130 to 210 g/kg diet. 5. Regression of Na+,K+-ATPase-dependent respiration against protein synthesis in each of intercostal and sartorius muscles showed a linear relation, an attestation of a close link between productive processes and auxiliary energy expenditure.

Animals↗

PubMed1980

Measurement of intracellular pH of bullfrog skeletal muscle and renal tubular cells with double-barreled antimony microelectrodes.

A pencil-type antimony microelectrode of double-barreled design with a tip of less than 1 to 2 microns in outside diameter was constructed and used to measure intracellular pH(pHi) on frog sartorius muscle and renal tubular cells. Simultaneous observations of membrane potential difference (EM) were made. The results obtained were as follows: (1) The in vivo pHi of frog sartorius muscle was 7.12 +/- 0.07 (SD) (n = 144); the simultaneously measured EM was -51.1 +/- 7.9 mV. The in vivo pHi of frog proximal tubule was 7.49 +/- 0.07 (n = 221) and the EM Peri across the peritubular membrane was -50.2 +/- 9.0 mV. (2) In proximal tubule in vivo, there was a negative correlation between pHi and EM (r = -.62, p < .05). On the other hand, in sartorius muscle in vivo, a positive correlation between the two was found (r = .85, p < .001). (3) In in vitro sartorius muscle, the pHi was 7.03 +/- 0.14 (n = 9) and EM was -62.4 +/- 4.4 mV within one hour after isolation. (4) Increasing the external potassium concentration in the preparations to 75 mM caused a progressive depolarization by 43.3 +/- 15.9 (m = 4) mV, while pHi changed in the alkaline direction by 0.22 +/- 0.03 pH unit. (5) These results indicate that the pHi in both tissues does not obey the Donnan rule.

Animals↗

PubMed1981

Effect of acetylcholine on ion transport in the frog skeletal muscle.

1. The effect of acetylcholine (ACh) on the ion transport of frog (Rana esculenta) sartorius muscles was studied. ACh was applied in bathing solution, Na influx and K efflux were measured using 24Na and 42K isotopes. 2. Na influx of sartorius muscles was increased by 1 mmol/1 ACh 2-10 fold depending on the experimental arrangement. The increase was greater if Na influx was measured at the beginning of ACh depolarization. During ACh treatment the Na influx took about the same time course as the depolarization recorded extracellularly. This type of recording approximately reflects the depolarization proceeding on the sartorius muscle fibres. 3. The presence of 31 nmol/l tetrodotoxin (TTX) did not modify the degree of increase of Na influx. 4. Rate coefficients for K efflux were increased 2-5 fold by ACh. The maximum rate coefficients were obtained in the first minute of ACh treatment. 5. Increase in K loss evolves also in the presence of 31 nmol/l TTX. The increase in rate coefficients was found to be about 30% less than without TTX in the first minute of ACh action. 6. The results indicate that in the presence of ACh the observed increase in Na influx and K efflux is brought about mainly by changes in Na and K conductance induced by ACh at the end-plates rather than by the action potentials accompanying ACh depolarization.

Acetylcholine↗