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Resolution of the potassium ion pump in muscle fibers using barium ions.

When frog sartorius muscles recover from Na enrichment in the presence of external K, net K entry into the fibers occurs by both passive movement and active inward transport via a K pump. Under normal conditions, it has not been possible to experimentally distinguish these processes. Fractionation into the flux components must be accomplished from inferences concerning the K conductance or permeability during a period of rapid Na extrusion. The best estimates indicate that 60-80% of the K entry occurs via the K pump. In the presence of Ba ions, the membrane permeability to K is very much reduced. Under these conditions, Na-enriched muscles underwent a normal recovery in the presence of external K, and the amount of inward K movement due to the K pump rose to over 90% of the total K entry. The characteristics of the K pump studied by this means were: (a) essentially complete inhibition by 10(-4) M ouabain, (b) inhibition by [Na]omicron, (c) activation by [K]omicron according to a rectangular hyperbola in the absence of [Na]omicron, (d) linear activation by [Na]iota over a wide range in concentration, (e) zero or undetectably low pumping rate as [Na]iota leads to 0, (f) the number of Na ions actively transported per K ion actively transported is 1.4-1.7 normally and 1.1 in the presence of Ba.

Animals↗

Acetylcholine synthesizing enzymes in frog skeletal muscle.

Acetylcholine synthesis in homogenates of frog sartorius muscle was measured by a radiometric method with a low blank. Choline acetyltransferase activity was very low (Vmax, 2nmol . g-1 . h-1, Km for choline, approx. 50 microM). The enzyme was found only in the endplate area and disappeared after denervation; it was inactivated by 4-(1-naphthylvinyl)pyridine. At high substrate concentrations its activity was overshadowed by the acetylcholine-synthesizing activity of a different enzyme not saturated by 10 mM-choline. The non-specific enzyme was present at and away from the endplate area, and it was not affected by denervation.

Acetylcholine↗

Transport of caesium in frog muscle.

1. The entry of caesium into sartorius muscle cells is strongly suppressed by the presence of 10(-5)M strophanthidin in Ringer solution.2. The amount by which caesium entry is reduced in the presence of strophanthidin is dependent on the intracellular sodium concentration and is greater the higher the intracellular sodium concentration.3. The magnitude of caesium influx in the absence of strophanthidin is highly dependent on the intracellular sodium concentration.4. Caesium uptake by muscles in which sodium has been largely replaced by lithium is reduced to very low values.5. Caesium can promote the extrusion of sodium from muscles with high intracellular sodium concentrations. The effects of 25 mM caesium and 5 mM potassium on sodium extrusion are roughly the same.6. External potassium inhibits the entry of caesium ions into muscle cells presumably by competing for transport sites.7. The drug strophanthidin has no effect on (134)Cs efflux provided that muscles have been loaded with tracer ions for a long period of time. Caesium efflux from the intracellular compartment appears to occur by a process not mediated by metabolism.8. The action of strophanthidin on (134)Cs efflux from muscles exposed to tracer for short times suggests that caesium ions are transported inwardly by an active process after first accumulating in a superficial reservoir.

Animals↗

Hydration effects on muscle response.

The effect of bathing toad sartorius muscle in hypotonic solutions is studied. The twitch, tetanus force and active stiffness increase as the tonicity decreases from 1.0 R to 0.6 R and then decrease below 0.6 R. Explanation is given in terms of the effect on ATP-ase activity.

Animals↗

The sarcoplasmic reticulum and transverse tubules of the frog's sartorius.

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.

Animals↗

Free anteromedial thigh flap: clinical application and review of literature.

The anteromedial thigh (AMT) flap is reviewed in terms of its vascular anatomy and previous clinical reports in the literature. Our own series of 5 patients treated with this flap for defects in the head and neck region and lower extremity is presented. Although several authors controversially discussed vasculature, we constantly found the pedicle as an emerging septocutaneous perforator at a point where the medial border of the rectus femoris muscle is crossed by the sartorius muscle. In all 5 patients, the AMT flap provided stable coverage with no flap loss. Based on our findings, we conclude that the anteromedial thigh flap offers all the advantages of fasciocutaneous flaps. Therefore, we recommend this flap as an alternative for defects requiring coverages of thin to moderate skin thickness. However, it should be remembered that variations in vascular anatomy are possible.

Aged↗

The structure of segments of the anisotropic band of muscle. II. Preparation and properties of A segments from vertebrate skeletal muscle.

The anisotropic band of skeletal muscle is a complex structural assembly of the protein myosin and associated nonmyosin components. To study the relationships among these proteins, aggregates of thick myofilaments held together at the M line (A segments) have been prepared from fresh and glycerol extracted chicken pectoralis and rabbit psoas muscles and from fresh frog sartorius muscle. The structure of the A segments included several thick filaments, an M line, and a bare zone or pseudo-H zone, lateral to the M line. Most of the A segments exhibited a pattern of eleven periodic stripes in each half lateral to the bare zone. The A segments from fresh muscle displayed these stripes more consistently than did the A segments from glycerinated muscle. Some of the major stripes appeared to be double, and there were two subdivisions between the stripes nearest the bare zone. The more lateral of the major A band stripes, however, had one subdivision between them. The M line consisted of three prominent medial stripes and two fainter lateral stripes. In the M lines of rabbit A segments the lateral stripes were located well into the bare zone whereas the lateral stripes of M lines in chicken A segments were closer to the three medial M line stripes. Our results on the preparation and properties of A segments are compared with those of of the investigators.

Animals↗

Diazepam blockade of repetitive action potentials in skeletal muscle fibres. A model of its membrane action.

Diazepam inconcentrations of 10(-6) and 10(-5) M effectively blocks trains of action potentials (AP) in individual rat diaphragm muscle fibres 'in vitro'. Single APs, evoked and recorded intracellularly with double microelectrodes, were not affected by diazepam. Trains of APs, evoked in diaphragm fibers in a SO42-- -medium (Cl-- -free), were not affected by diazepam. Diazepam decreased the dependence of the membrane resting potential of muscle cells in chloride media on elevated external potassium whereas no effect was observed in SO42-- --Cl-- -free saline. Similar to in the rat diaphragm, diazepam blocked the trains of AP of individual muscle fibres in the frog sartorius muscle (Cl-- -medium). Diazepam did not affect the spontaneous fibrillatory APs recorded extracellularly from rat gastrocnemius muscles 10 days after denervation. Diazepam apparently increases the resting permeability of the muscle fibre memebrane for chloride ions. In the presence of the drug, the higher permeability for Cl- diminishes the depolarization caused by the potassium released and accumulated in the vicinity of the membrane in the course of AP. The muscle fibre membrane thus does not respond by repetitive activity when partially depolarized and only one AP can be evoked when diazepam is present. It is suggested that similar changes in Cl-- permeability may occur in brain excitable structures after diazepam administration and may account for some of the therapeutic effects of the drug.

Action Potentials↗

Anti-curare action of stannous ion in the frog neuromuscular junction.

For the purpose of elucidating the mechanism of action of stannous ion (Sn2+), we investigated effects of stannous chloride (SnCl2) on the twitch and on the electrical phenomena in the muscle fiber. Sciatic nerve-sartorius muscle preparations from the bullfrog were used as the material. Effect of SnCl2 was examined on the twitch partially inhibited by pretreatment with d-tubocurarine. SnCl2 (1-100 microM) antagonized d-tubocurarine and enhanced the twitch dose-dependently. Tartaric acid, which is the solvent used for SnCl2 solution, had no augmentative effect on the twitch, even at a concentration as high as 250 microM. SnCl2 (1-50 microM) increased the amplitude of the endplate potential; that is, it exerted an anti-curare action. The resting potential and the membrane resistance of the muscle fiber were not altered by 30 microM SnCl2. These findings lead to the conclusion that Sn2+ enhances the twitch by increasing the endplate potential of the muscle fibers.

Animals↗

Myotonia as a side effect of diuretic action.

1. Commonly used loop diuretics produce side effects in man which are similar to chemically induced myotonia. These diuretics have structural affinity with known myotonic agents. 2. We have observed EMG myotonia in vivo in leg muscles of rats treated with intravenous frusemide. 3. In the presence of several different diuretics, rat isolated diaphragm, soleus and extensor digitorum longus muscles as well as frog sartorius muscles produce typically myotonic contractions with relaxation times up to several seconds. 4. Intracellular recording of action potentials from diuretic-treated muscles reveals long lasting after-discharges following a brief electrical stimulus, again typical of chemically induced myotonia. 5. Having demonstrated a myotonic action of several diuretics we suggest a need for caution in using these drugs in persons with hereditary myotonia and a need to be aware of possible provocation of myotonia in subclinical cases. Myopathies and neuropathies which are known to result from chronic exposure to myotonic agents also need to be considered. 6. In our study, the diuretic, acetazolamide, unmasked subthreshold myotonia. This seems to be at variance with reports of its usefulness in the treatment of myotonia. 7. Diuretics should probably not be employed in the treatment of herbicide intoxication where their myotonic activity would be expected to add to the known myotonic activity of the herbicide.

Animals↗

Structural and functional asymmetries of myelinated branches in the frog muscle spindle.

1. Most of the axons innervating single-type muscle spindles in the frog sartorius muscle divide into myelinated branches at a node; one of the branches subdivides (Bs) and the other does not (Bn).2. Electrical responses at the afferent nerve terminal of the isolated spindles were recorded extracellularly and intracellularly with micro-electrodes, concurrently with the terminal responses by the paraffin gap method.3. By recording spontaneous orthodromic and provoked antidromic impulses and by stimulating the nerve terminals with micro-electrodes and inactivating the terminals with micro-application of potassium ion, it was deduced that propagated and abortive spikes are almost always generated at the terminal of Bn (Tn), not at the terminals of Bs (Ta and Tb).4. The site of impulse initiation, as determined in (3), was found, after measurement of the lengths of the myelinated branches in each preparation, to be essentially the terminal nearest the parent axon.5. Selective inactivation of Tn by far-ultra-violet light removed both propagated and abortive spikes. Both types of spikes survived after irradiation of Ta and Tb, although the relationship between spike frequency and spindle stretch was altered. The same results were obtained after transection of the branches with a laser microbeam.

Animals↗

[Effects of three Chinese traditional drugs on afferent discharge from single muscle spindle in toads].

OBJECTIVE: To shed some light on treatment of muscle atrophy through testing the effects of several Chinese traditional drugs that can invigorate blood circulation on the afferent discharge of muscle spindle. METHOD: Single muscle spindle was isolated from sartorius muscle of toad. Afferent discharge from the muscle spindle was recorded using air-gap technique and effects of three Chinese traditional drugs on afferent activity of the muscle spindle were observed. RESULT: Ligustrazine could markedly tingle the spontaneous electrical activity of the muscle spindle, and it also showed a good dose-response relationship, while Salvia Miltiorrhiza and Safflower had no such excitative effect. CONCLUSION: It is known that ligustrazine can expand blood vessels and agitate microcirculation, and this effect on the muscle spindle makes it helpful for persons with muscle atrophy.

Animals↗

Structural changes in myosin cross-bridges during shortening of frog skeletal muscle.

X-ray diffraction patterns from frog sartorius muscle were recorded during steady shortening with various loads. The intensity of the third meridional reflection from the thick filament decreased on shortening to an extent proportional to the drop in tension. The intensity correlated more closely with the tension than with the shortening velocity. The Bragg spacing of the third meridional reflection decreased in proportion to the decrease in tension. The intensity decrease of the actin layer lines at 1/5.1 and 1/5.9 nm-1 was roughly proportional to the decrease in the load, indicating that the number of cross-bridges decreases similarly. The intensity of the (1,1) equatorial reflection showed a significant decrease only with low loads. Assuming that a steady structural state is attained during steady shortening, the results are consistent with the cross-bridge model in which the number of myosin cross-bridges decreases during shortening.

Actins↗

X-ray diffraction observations of chemically skinned frog skeletal muscle processed by an improved method.

Whole frog sartorius muscles can be chemically skinned in approximately 2 h by relaxing solutions containing 0.5% Triton X-100. The intensity and order of the X-ray diffraction pattern from living muscle is largely retained after such skinning, indicating good retention of native structure in fibrils and filaments. Best X-ray results were obtained using a solution with (mM): 75 K acetate; 5 Mg acetate; 5 ATP; 5 EGTA; 15 K phosphate, 2% PVP, pH 7.0. Equatorial X-ray patterns showed that myofibrils swell after detergent skinning, as also observed after mechanical skinning. This swelling could be reversed by adding high molecular weight colloids (PVP or dextran) to the extracting solution. By finding the colloid osmotic pressure needed to restore the in vivo interfilament spacing (3% PVP, 4 X 10(4) mol wt) the swelling pressure was estimated as 35 Torr in a standard KCl-based relaxing solution. The swelling pressure and the extent of swelling were less than acetate replaced chloride as the major anion. Detergent-skinned muscle lost the constant-volume relation between sarcomere length and lattice spacing seen in intact muscle. Changes in A band spacing were paralleled by changes in I and band-Z line spacing at a constant sarcomere length. After detergent skinning, I1,0 rose while I1,1 fell, a change in the relaxing direction. Since raising the calcium ion concentrations from pCa 9 to PCa 6.7 was without effect on equatorial or axial X-ray patterns, we concluded that these intensity changes were not due to calcium-dependent cross-bridge movement but rather to disordering of thin filaments in the A band.

Animals↗

Muscle contraction generates discrete sound bursts.

Isolated frog sartorius muscles were stimulated to shorten under lightly loaded conditions. A piezoelectric transducer was placed alongside the muscle to record sounds generated during contraction. Shortening was accompanied by the generation of a series of discrete sound bursts. The bursts were found to be moderately repeatable among successive contractions; 44% repeated from contraction to contraction. The duration of each sound burst was on the order of 400 mus, and the temperature dependence of the interval between successive bursts had a Q10 of approximately 2. Sound intensity was variable: average acoustic power ranged from 0.05-0.4 mW/g, or approximately 1% of the heat generated during contraction. The generation of discrete bursts of sound during contraction, rather than continuous sound, implies that contractile behavior may be discontinuous.

Animals↗

Structure-activity relationships of several cardiotonic steroids with respect to inhibition of ion transport in frog muscle.

Cesium uptake by sodium-loaded frog sartorius muscles was inhibited 100% by 10(-6)M ouabain and 10(-6)M cymarin. The doses for 50% inhibition of cesium uptake by five cardiotonic aglycones were 1.5 x 10(-6)M for strophanthidin, 2 x 10(-7)M for telocinobufagin, 1.6 x 10(-6) for digitoxigenin, 2.4 x 10(-6)M for periplogenin, and 6.3 x 10(-6)M for uzarigenin. Because of the limited solubility of sarmentogenin the maximum concentration studied was 2 x 10(-6)M which inhibited cesium uptake about 36%. Inhibition of cesium uptake by cymarin was not reversed during a 3.5 hr incubation in fresh solution while the muscles treated with ouabain and strophanthidin recovered partly during this time. Cymarin was a more potent inhibitor of sodium efflux than strophanthidin and periplogenin was less potent. Increased cesium ion concentration in the external solution decreased the strophanthidin inhibition of cesium uptake but 25 mM cesium did not overcome the inhibition by 10(-8)-10(-6)M strophanthidin. Increased potassium ion concentration in the external solution decreased but did not completely overcome inhibition of sodium efflux by strophanthidin. It is concluded that potassium or cesium ions do not compete with these drugs for a particular site on the ion transport complex. The same structural features of the drugs are necessary for inhibition of ion transport in frog muscle as are required for inhibition of ion transport in other tissues, inhibition of sodium-potassium-stimulated ATPases, and toxicity to animals.

Animals↗

Transient phases of the isometric tetanus in Frog's striated muscle.

In an isometric tetanus in frog's sartorius muscle tension approaches the plateau exponentially with rate constant alpha. alpha a depends on sarcomere length, s, and temperature, T, according to the Arrhenius equation See PDF for Equation for temperatures between 1 and 20 degrees C and for sarcomere lengths 2.0-2.8 microm. The energy of activation, E, does not vary significantly with s; E = 13.9 +/- 2.4 kcal/mole. A(s) decreases monotonically with s; A(2.1 microm) is about three times greater than A(2.8 microm). Late in relaxation active tension approaches zero exponentially with rate constant r. r decreases exponentially with increasing duration of tetanus, D, from r(0) in a twitch to r(infinity) for large D. The rate constant for decrease of r with D increases with s and with T. r(0) and r(infinity) obey the Arrhenius equation and decrease with increasing s.

Animals↗

Tension due to interaction between the sliding filaments in resting striated muscle. The effect of stimulation.

1. A resting sartorius muscle of the frog or toad possesses a special kind of elasticity which is shown to be due to a component lying between the two sets of filaments. The elastic effect is seen only for very small length changes, up to about 0.2% of the muscle length, and the ;elastic limit' is then reached. If the length change then continues at a constant velocity the tension developed is maintained at a fixed level, producing a sort of frictional resistance. The component responsible is called the ;short-range elastic component', or SREC.2. It is also shown that a small part of the permanent tension of a resting muscle is probably due to ;active' interaction between the filaments. This is called the ;filamentary resting tension', or FRT. For a sarcomere length of 2.0 mu the FRT amounts to about 150 mg in a muscle weighing 100 mg.3. The stiffness of the SREC and the magnitude of the FRT are shown to be related to one another. They are both increased, and may rise to high values, by making the external solution hypertonic.4. The working hypothesis is as follows. In a resting muscle the cross-bridges on the myosin filaments are not entirely inactive, but a very small proportion of them are cross-linked with the actin filaments. The links are very stable and have a long ;life'. The elastic behaviour is due to the flexural rigidity, or spring-like properties of these bridges. The elasticity is ;short-range' because the bridges can be bent, or stretched, only a small way from the steady-state position before the contacts ;slip'. The ;filamentary resting tension' (which is present in the absence of any external length change) is attributed to an ;active process', which operates by imparting ;organized' potential energy to the participating cross-bridges, by potentiating their attachment, against their elastic resistance, to sites on the actin filaments which are displaced towards the Z line.5. It is shown that the ability of the muscle to maintain a ;frictional' resistance to a continuing, slow, length change is suppressed during a maintained tetanic contraction. The process of suppression starts during the period of the latency relaxation.6. It is suggested that the latency relaxation may be due to a reduction of the filamentary resting tension.7. The experiments with the latency relaxation were facilitated by the use of strongly hypertonic solutions. The positive twitch tension is then greatly reduced, or may be practically abolished, while the latency relaxation remains at about its normal size and is extended in duration.

Animals↗