Neurotrophic control of the transmembrane Cl− pump in mammalian muscle fibers

Resting membrane potential of both innervated and denervated rat diaphragm muscle fibers was investigated when the concentration of potential-producing ions was changed in the external fluid and following treatment with furosemide. It was found that equilibrium potential ( ) equalled resting potential level in innervated muscle for Cl^–, but shifts to more positive values compared with resting membrane potential following section of the nerve. Furosemide retards development of post-denervation depolarization of the muscle membrane. It is deduced that trophic influences originating from the motor nerve activates the furosemide-sensitive Cl^– influx system, leading to raised intracellular concentration of Cl^–, a shift in (E_Cl) and depolarization of the muscle membrane.S. V. Kurashov Medical Institute, Minsitry of Health of the RSFSR, Kazan'. Translated from Neirofiziologiya, Vol. 19, No. 6, pp. 766–771, November–December, 1987.     

The ATP/ADP-antiporter is involved in the uncoupling effect of fatty acids on mitochondria

The ATP/ADP-antiporter inhibitors and the substrate ADP suppress the uncoupling effect induced by low (10-20 microM) concentrations of palmitate in mitochondria from skeletal muscle and liver. The inhibitors and ADP are found to (a) inhibit the palmitate-stimulated respiration in the controlled state and (b) increase the membrane potential lowered by palmitate. The degree of efficiency decreases in the order: carboxyatractylate (CAtr) greater than ADP greater than bongkrekic acid, atractylate. GDP is ineffective, Mg.ADP is of much smaller effect, whereas ATP is effective at much higher concentration than is ADP. Inhibitor concentrations, which maximally suppress the palmitate-stimulated respiration, correspond to those needed for arresting the state 3 respiration. The extent of the CAtr-sensitive stimulation of respiration by palmitate has been found to decrease with an increase in palmitate concentration. Stimulation of the controlled respiration by p-trifluoromethoxycarbonylcyanide phenylhydrozone (FCCP) and gramicidin D at any concentrations of these uncouplers is CAtr-insensitive, whereas that caused by a low concentrations of 2,4-dinitrophenol and dodecyl sulfate is inhibited by CAtr. The above effect of palmitate develops immediately after addition of the fatty acid. It is resistant to EGTA as well as to inhibitors of phospholipase (nupercain) and of lipid peroxidation (ionol). Moreover, palmitate accelerates spontaneous release of the respiratory control, developing in rat liver mitochondria under certain conditions. This effect takes several minutes, being sensitive to EGTA, nupercain and ionol. Like the fast uncoupling, this slow effect is inhibited by ADP but CAtr and atractylate are stimulatory rather than inhibitory. In artificial planar phospholipid membrane, palmitate does not increase the membrane conductance, FCCP increases it strongly and dinitrophenol only slightly. In cytochrome oxidase proteoliposomes, FCCP, gramicidin and dinitrophenol (less effectively) lower, whereas palmitate enhances the cytochrome-oxidase-generated membrane potential. In this system, monensin substitutes for palmitate. It is concluded that the ATP/ADP antiporter is somehow involved in the uncoupling effect caused by low concentrations of palmitate and, partially, of dinitrophenol, whereas uncoupling produced by FCCP and gramicidin is due to their action on the phospholipid part of the mitochondrial membrane. A possible mechanism of this effect is discussed.     

A method for determining the toxigenicity of micromycetes]

A method is suggested to determine toxigenicity of microscopic fungi from genera Aspergillus, Penicillium, Fusarium, etc. which contaminate fodder, food products and environment objects. The method is based on the property of micromycetes isolated from fodder, food products and environment objects to produce mycotoxins under optimal conditions of their cultivation on the nutrient media. Their further extraction by the organic solvent, evaporation and emulsification of the extract and its peroral introduction to young female rats cause a disease and death of animals. The suggested method may be used for recognizing and preventing mycotoxicoses of farm animals and poultry.     

NMR study of the structural transition in the DNA-bound water system in the interval of physiological temperatures

Temperature dependence of spin-spin proton relaxation times of DNA and bound water and the content of bound water in the samples of DNA, saturated with water in the atmosphere with different relative humidities from 0 to 100% were studied by means of pulsed NMR. It is shown that the temperature transition in the system of DNA-bound water in the interval 18-35 degrees is observed only when the relative humidity is more than 70% and the double-stranded structure of DNA exists. The transition of DNA from one conformation into another passes through some intermediate state more labile and probably less ordered. This transition is accompanied by changes in the structure of the hydration shell. In the case when relative humidity is greater than 80%, the partial dehydration of DNA stimulated by the transition is observed. This dehydration increases with the increase of relative humidity.     

Modification of the lipid composition of neuroblastoma C1300 cells using liposomes

An essential increase in the total cholesterol content with a significant growth in the level of its esters was observed in neuroblastoma C1300 cells differentiated by means of 5'-bromodesoxyuridine and incubated for 1 h with lecithin-cholesterol liposomes (1:1 mol/mol). These cells also displayed an increased amount of polyunsaturated fatty acids in the composition of lecithin molecules and the appearance of lysolicethin as well. Incubation of the cells with lecithin liposomes was accompanied by a decrease in the content of cholincholesterol in the cells. In this case the level of cholesterol esters decreased to a greater extent than the level of free cholesterol. In lecithin molecules of these cells there occurred a relative increase of saturated fatty acids. The cells can retain viability and compensate changes caused by cholesterol excess during their incubation with lecithin-cholesterol liposome for 60-90 min. A longer incubation with liposomes results in a sharp drop of cell viability.     

Membrane model of the regulation of proliferation: the theory and interpretation of an experiment

The role of cell surface physical organization in the cell cycle regulation is analyzed within the framework of the earlier proposed theory (Chernavskii et al., 1982). Two models of cell surface are considered: hard-frame fluid-mosaic model (latticemosaic) and the fluid-mosaic one. The former deals with normal cells. The existence of integral carcasse or "frame" which is formed by the essential part of cross-linked membrane components and may have at least two different conformational states is hypothesized. The second model describes membranes of tumour cells. With the latter theory any mitogen (excluding the restoration of nutrient depletion) reduces the mechanical tensile strength of the frame and stimulates the general structural rearrangement of the plasma membrane. There are only two conformational transitions during the cell cycle which serve as signals for the beginning of S and M phases. If the values of tensile strength are great enough and therefore the conformational transitions are impossible, the cells pass into the resting (prereplicative--G01, or premitotical--G02) state. Three types of experiments are interpreted in the proposed theory: a) on differences in the action of growth factors on normal and tumour cell cycle, b) on the necessary condition for mitogenicity of lectins, c) on the stimulation of proliferation by mechanical deformation of cells.     

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.     

Effects of hyperosmolarity and furosemide on resting membrane potentials and skeletal muscle fiber volume in rats

The changes of the muscle fibres volume and resting membrane potential (RMP) were studied following treatment with hypertonic medium and furosemide. The volume changes in hypertonic medium began with cell shrinkage and later have been followed by the volume increase up to normal level during 30-40 minutes. At the same time the medium hypertonicity caused muscle fibres depolarisation. The hypertonic-induced decrease of the RMP was delayed in the furosemide-treated muscle. Besides, furosemide abolished the muscle fibres volume restorative properties in hypertonic medium. It is suggested that the membrane depolarisation and cell volume restoration in hypertonic medium are the resultant effects of intracellular chloride ions level elevation which, in turn, have been evoked by activation of furosemide-sensitive Cl(-)-influx system.