Effect of postsynaptic blockade of neuromuscualar transmission on properties of the frog fast muscle fiber membrane

Sensitivity of the postsynpatic membrane to acetylcholine, the resting membrane potential, input resistance, and membrane time constant of fast muscle fibers were measured in experiments on frogs. Complete immobilization of the animals with D-tubocurarine or local immobilization of a muscle with α-bungarotoxin was found not to affect these parameters of the muscle membrane, whereas denervation of the muscle widens the zone of postsynaptic sensitivity to acetylcholine, lowers the resting membrane potential, and increases the input resistance and time constant of the muscle membrane. These results are evidence that neurotrophic control of the frog fast muscle fiber membrane is achieved mainly by substances reaching the muscle via axoplasmic transport and not by the character of the neuronal discharge and motor activity or by synaptic acetylcholine.     

Nonquantum release of acetylcholine in frog neuromuscular junction after blocking of axoplasmic transport with colchicine

Standard microelectrode techniques were used to evaluate the effect of d-tubocurarine chloride on membrane potential of junctional and extrajunctional areas of muscle fibers in a potassium-free Ringer solution. The experiments were made on frogs after inactivation of acetylcholinesterase. d-Tubocurarine chloride hyperpolarized the membrane of muscle fibers only in the junctional area. Blockade of axoplasmic transport with colchicine did not affect the magnitude of the hyperpolarization response of the membrane end plate to the presence of d-tubocurarine chloride, but at the same time it significantly reduced the membrane rest potential of muscle fibers, and gave rise to the appearance of extrajunctional sensitivity to acetylcholine. It is concluded that the blockade of axoplasmic transport does not affect the pattern of non-quantum acetylcholine release from nerve terminals. Therefore, this is unlikely to cause denervation-like changes in the muscle under the conditions described.     

Electrical properties of normal and dysgenic mouse skeletal muscle in culture

Electrical properties of normal and dysgenic mouse skeletal muscle were studied by intracellular recording from embryonic cells developing in vitro. Passive membrane constants were determined from records of transmembrane potential responses to hyperpolarizing pulses of current using two types of analyses, assuming the tubes to be finite cylinders: the off transient and steady state analyses. The following properties of normal and dysgenic fibers were also studied. (a) membrane potentials (b) acetylcholine sensitivity (c) α-Bungarotoxin binding and (d) maximum rate of rise, overshoot and one-half fall time of the action potential. Rare electrotonic coupling between fibroblasts and myotubes was noted. An anomalous type of rectification Was observed in some fibers in which the transmembrane potential responses possessed under and overshoots. These responses may have affected the values of membrane constants as derived by the off transient analysis. In all parameters studied, including membrane constants derived by the steady state analysis, the cultured mouse cells resembled adult denervated mammalian muscle rather than innervated muscle. There were no differences between normal and dysgenic fibers with respect to any of the parameters studied. Dysgenic fibers did not contract although they displayed passive and active membrane properties like those in normal, non-dysgenic fibers.     

Electrical and acetylcholine-receptive properties of the muscle fiber membrane after axoplasmic transport blockade by colchicine

Electrical properties of the membrane and sensitivity of the fibers to acetylcholine were investigated in the frog sartorius muscle after denervation and a single application of colchicine to the nerve. After both types of procedure the electrical properties showed similar changes and extrasynaptic sensitivity to acetylcholine appeared. No such changes took place in the fibers of the contralateral muscle. Injection of colchicine into the lymphatic sac did not affect the electrical properties of the membrane, but widened the zone of sensitivity to acetylcholine. The results are regarded as further evidence in support of the view that denervation-like changes after application of colchicine to the motor nerve, when the transmission of excitation of nerve to muscle is preserved, are the result of a disturbance of the supply of neurotrophic substances along the axon by means of axoplasmic transport.Kazan' Medical Institute. I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 12, No. 5, pp. 550–557, September–October, 1980.     

Role of opioid peptides in the neurotrophic control of the acetylcholine-sensitive membrane of rat skeletal muscle fibers

The influence of opiate peptides on the development of hypersensitivity to acetylcholine has been revealed in muscle fibers membrane after denervation of rat diaphragm muscle. The addition of 1 X 10(-8) M beta-endorphin or dalargin to the culture medium prevented the appearance of extra-junctional acetylcholine sensitivity. The peptide containing only three amino acids and identical to the initial dalargin region did not possess the same effect. Moreover, if this peptide was present in the culture medium in the concentration higher than that of dalargin, the effect of dalargin was blocked. It is suggested that the neurotrophic regulation of acetylcholine-sensitive membrane properties of skeletal muscle fibers is accomplished with opiate neurogenic peptide, e.g. beta-endorphin.     

Membrane currents of rat satellite cells attached to intact skeletal muscle fibers

Muscle satellite cells play an important role in the postnatal growth of skeletal muscle and in the regeneration of damaged muscle during adult life. Little is known about the physiological properties of satellite cells in their dormant state as they lie adjacent to the intact muscle fibers, underneath the basement membrane. Our recent experiments, using patch clamp techniques, indicate that no tight electrical coupling is present between satellite cells and the muscle fiber dissociated from rat flexor digitorum brevis. Satellite cells possess sodium channels with low sensitivity to tetrodotoxin and at a much lower density than muscle. In addition, satellite cells are insensitive to acetylcholine (ACh) for at least 24 hr after having been removed from the animal, even when detached from their muscle fiber. However, we could measure ACh-evoked currents from satellite cells 48-72 hr in culture, indicating that ACh sensitivity develops with time.     

Effect of nicotine and acetylcholine on crustacean muscle membrane potential

We have investigated the effect of nicotine and acetylcholine on the resting membrane potential of the crayfish extensor muscle in order to determine whether crustacean muscle can be activated by cholinergic compounds. Intracellular recordings from individual deep extensor abdominal muscle cells were made using standard glass microelectrode techniques. The resting membrane potential was measured before and after treatment with glutamate, nicotine, and acetylcholine. Glutamate, which is a known activator of crayfish muscle, was used to determine whether the muscle cell preparation was viable and capable of responding to any of the test substances. Our results confirm that application of glutamate is associated with a depolarization of the muscle membrane. However, muscle cells showed no depolarization after treatment with nicotine (50 microM) or acetylcholine (66 microM). These results argue against the notion that increases in muscle tension may be responsible for the increased receptor organ discharge observed in the presence of nicotine. Rather, it supports the hypothesis that nicotine is acting directly on the mechanoreceptor membrane to change its sensitivity.     

Tenotomy of the avian anterior latissimus dorsi muscle. II. Can regeneration from the stump occur in the pigeon?

Because the chick's anterior latissimus dorsi muscle (ALD) regenerates a fast-twitch muscular connection after tenotomy, the pigeon's ALD was tenotomized, either at the origin or through the muscle 0.5 cm from the origin, to determine whether this muscle behaves similarly to the chick muscle. These procedures were compared in pigeons operated upon at 7 weeks, versus 5 to 9 months of age. The pigeon's ALD did not regenerate a new connection, and other differences were observed between the pigeon and chick ALD. The pigeon ALD has only a single slow muscle-fiber type, has fewer fast fibers, and transforms to a fast-twitch muscle more readily than the chick ALD after tenotomy. The transformation of muscle fiber types occurred more readily in the older pigeons than those tenotomized at 7 weeks of age. Tenotomy induced morphological alterations of the muscle fiber structure in all of the pigeons, which is in contrast to the absence of changes in the tenotomized chick ALD. Therefore the pigeon and chick ALD respond completely differently to tenotomy.     

Daily muscle vibration amelioration of neural impairments of the soleus muscle during 2 weeks of immobilization

V-wave, F wave and H-reflex responses of soleus were used to determine neural adaptations to 2-week immobilization and whether muscle vibration intervention during immobilization would attenuate the negative adaptations induced by immobilization. Thirty subjects were divided into the ankle immobilization group and the immobilization with muscle vibration group. Mechanical vibrations with constant low amplitude (0.3 mm) were applied (12 × 4 min daily) with a constant frequency of 100 Hz on the soleus muscle of the subjects in vibration group during the ankle immobilization period. Soleus maximal M-wave (Mmax) and H-reflex (Hmax) were evoked at rest. F-wave was recorded by supramaximal stimulation delivered at rest and V-wave during maximum voluntary contraction (MVC). The EMG during MVC was represented by its root-mean-square (RMS) value. Each subject was examined before and after 2 weeks of immobilization. Results showed that following 2 weeks of immobilization, Mmax, Hmax and F wave all did not change with immobilization in either group (P > 0.05). After 2 weeks of immobilization, significant reductions in V/Mmax (of 30.78%) (P < 0.01) and EMG RMS (24.82%) (P < 0.001) were found in the immobilization group. However, no significant changes occurred in the immobilization with muscle vibration group. Such findings suggested that 2 weeks of immobilization resulted in neural impairments as evidenced by the reduction in EMG and V wave, and that such decrease was prevented by the intervention of muscle vibration during the immobilization period.     

Effect of denervation and tenotomy on slow and fast muscles of the chicken

Summary Changes of muscle weights, fiber diameters and ultrastructure were studied in the slow anterior latissimus dorsi (ALD) and in the fast posterior latissimus dorsi (PLD) of the chick three weeks after denervation and tenotomy, and after combined denervation and tenotomy of the two muscles.The slow ALD muscle becomes hypertrophic after denervation (Feng, Jung and Wu, 1962). Three weeks after nerve section, wet weights of ALD muscles are increased by 60% and fiber diameters become by 30% larger than those of contralateral control muscles. In spite of this hypertrophy, degenerative changes are seen in the ultrastructure, similar to those described in denervated atrophic muscles. Areas of dedifferentiation with autophagic vacuoles and aggregates of tubules are found in superficial layers of some fibers. Disintegration of Z lines and filaments along one or two sarcomeres occurs in a number of myofibrils, especially in muscles of young animals.In contrast to denervation alone, simultaneous denervation and tenotomy of the ALD muscles results in atrophy. Decrease of muscle weights and reduction of fiber diameters are similar as after tenotomy; in both cases muscle fibers waste by degeneration and atrophy of myofibrils.The fast PLD muscles underwent extensive atrophy in all three series of experiments. Corresponding atrophic and degenerative changes of ultrastructure were found in all instances.The authors wish to acknowledge gratefully the skillful technical assistance of Mrs. M. Sobotková and Ing. M. Doubek, and editorial assistance of Miss Virginia Hamilton.     

Has nonquantal acetylcholine secretion from motor nerve endings a role in neurotrophic control of resting membrane potential in rat muscle fibers?

The resting membrane potential of fibers of the rat diaphragm was measured by a microelectrode technique 3 h after division of the phrenic nerve and incubation in culture medium for 5 days after denervation. The membrane potential was recorded in synaptic regions of fibers close to (2–3 mm) and distant from (9–11 mm) the site of nerve division. The membrane potential of the synaptic region of the close fibers 3 h after denervation became smaller, whereas that of the synaptic region of distant fibers did not change relative to the control. Placing the muscle 3 h after denervation into medium with carbamylcholine (1·10^–8 M), cGMP (1·10^–4 M), or dibutyryl-cGMP (1·10^–6 M) led to hyperpolarization of the synaptic region of the close fibers but did not change the resting potential in the synaptic region of the distant fibers, and abolished differences between them. Five days after division of the nerve, incubation of the muscle in a solution with the above-mentioned substances did not affect the resting membrane potential. Nonquantal release of acetylcholine from motor nerve endings, assessed by the amplitude of hyperpolarization of the postsynaptic membrane, induced by application of curarine against the background of acetylcholine esterase inhibition, 3 h after denervation was identical in the synaptic region of the close and distant fibers and did not differ from the control. It is postulated that the postdenervation fall of membrane potential of rat muscle fibers is not due to disturbance of nonquantal secretion of acetylcholine from motor nerve endings.S. V. Kurashov Kazan' Medical Institute, Ministry of Health of the USSR. Translated from Neirofiziologiya, Vol. 17, No. 3, pp. 358–365, May–June, 1985.     

Dystrophin-related protein in the fetal and denervated skeletal muscles of normal and mdx mice

The amino acid sequence of the polyclonal antibodies we developed against the carboxyl terminus of the dystrophin-related protein, the putative gene product of B3 cDNA, had no homologous sequence to the dystrophin molecule except for two amino acids located at its ends for immunization. By immunohistochemical examination in C57B1/10ScSn and C57B1/10ScSn-mdx mice we found that the DRP was expressed on the surface membrane of fetal muscle fibers, was assembled at the neuromuscular junctions of the mature muscle fibers, and reappeared on the surface membrane of muscle fibers after denervation. Its localization was similar to that of the acetylcholine receptor, suggesting that DRP is one of the cytoskeletons which organize and stabilize the cytoplasmic domain of the acetylcholine receptor.     

Functional differentiation of the tail muscle fibers in Rana temporaria tadpoles

Studies have been made on changes in the electrical properties of muscle membrane and lipid content of two types of myotomal fibers in the tail of tadpoles during metamorphosis. It was shown that during premetamorphosis, peripheral and inner muscle fibers do not differ with respect to their effective resistance, time constant of the membrane and lipid content; the resting membrane potential is higher in the inner fibers. During further development of the tadpoles, differentiation of muscle fibers takes place, and to the beginning of the climax the inner fibers attain lower values of the effective resistance and time constant, as well as lower content of lipids in their sarcoplasm; the difference in the level of resting membrane potential between the peripheral and inner fibers increases. The data obtained suggest that the inner fibers may be referred to as fast ones, whereas the peripheral ones--as slow. These data also reveal specific features in neurotrophic regulation of functional properties of muscle fibers in tadpoles.     

Conversion of rat muscle fiber types. A time course study

Rats were used in this study to determine the time course of conversion of muscle fiber types. The right or left gastrocnemius muscle was removed thereby causing an overload on the ipsilateral soleus and plantaris muscles. The contralateral limb served as a control. The type II to type I fiber conversion was followed histochemically in the soleus and plantaris muscles for one to six weeks following surgery. Muscle sections were stained for myofibrillar actomyosin ATPase and NADH tetrazolium reductase. The type I population in the soleus muscle was 99.3% six weeks after synergist removal. The plantaris muscle underwent a two fold increase in the percentage of type I fibers after six weeks. Transitional fibers were prominent in the plantaris muscle and reached their peak at 4% (P less than 0.05) of the total population, four weeks after surgery.     

Effects of β-endorphin on the development of denervation changes in rat muscle fiber membrane

Recordings were made of post-denervation changes in resting potential and input resistance in muscle fiber membrane, as well as anode break, tetrodotoxin resistant action potentials, and asynaptic sensitivity to acetylcholine during experiments on cultured diaphragm muscle fiber isolated from rats. Addition of -endorphin to the culture medium prevented increase in the input resistance of muscle fibers and reduced development of asynaptic transmitter sensitivity in the membrane, but failed to change the ability of the denervated muscle membrane to generate anode break and tetrodotoxin-resistant action potentials. The effects of -endorphin were not abolished by naloxone, which itself had endorphin-like powers as measured by the indices used in this research. It is therefore suggested that -endorphin or like substances could be claimed as the neurotrophic factors responsible for controlling passive electrical properties of the muscle fiber membrane and contribute to regulating its acetylcholine sensitivity.S. V. Kurashov Medical Institute, Ministry of Public Health of the RSFSR, Kazan'. Translated from Neirofiziologiya, Vol. 19, No. 6, pp. 759–766, November–December, 1987.     

Enzyme histochemical alterations in human skeletal muscles after tenotomy and after spontaneous rupture of the tendon.

The authors studied the histochemical alterations of human skeletal muscles after tenotomy and after spontaneous rupture of the tendon. Both succinate dehydrogenase (in type I fibers), and myofibrillar ATP-ase (in type 2 fibers) activity was decreased in all injured muscles. In the intact antagonists and contralateral muscles alterations were not found. The creatine phosphokinase and aldolase activity were decreased also in injured muscles. The lactate dehydrogenase activity was various both in affected and in unaffected muscles. Two weeks or more after the injury of the tendon in injured muscles the number of type 1 fibers were decreased and therefore a mathematically significant type 2 fibre predominance occurred. Atrophy involve both type 1 and type 2 fibers, but type 1 fibre atrophy was more pronunced as type 2 fibre atrophy.     

Functional properties of locust locomotor muscles

The ultrastructure of the muscle fibers and the electrical constants and responses of the membrane to microapplication of L-glutamate and acetylcholine were investigated in the longitudinal flight muscle and the flexor tibiae ofLocusta migratoria migratorioides. The twitch flight muscle differs from the slower leg muscle in the smaller size of its sarcomeres and the lower values of the space attenuation factor of the electrotonic potential, time constant, and resistance of the membrane. Microapplication of sodium L-glutamate at strictly definite points of the fibers of both muscles evoked depolarization responses of the membrane. In experiments on normal and denervated muscle, during microapplication of acetylcholine, changes in the level of the membrane potential were never observed. It is concluded that L-glutamic acid is the excitatory mediator of the twitch and slow muscle systems of insects.I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 9, No. 5, pp. 532–538, September–October, 1977.     

Regulation of Protein Synthesis in Inactivated Skeletal Muscle: Signal Inputs,Protein Kinase Cascades,and Ribosome Biogenesis

Disuse atrophy of skeletal muscles is characterized by a significant decrease in the mass and size of muscle fibers. Disuse atrophy develops as a result of prolonged reduction in the muscle functional activity caused by bed rest, limb immobilization, and real or simulated microgravity. Disuse atrophy is associated with the downregulation of protein biosynthesis and simultaneous activation of protein degradation. This review is focused on the key molecular mechanisms regulating the rate of protein synthesis in mammalian skeletal muscles during functional unloading.