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One of the strategies to induce tolerance of neurons to toxic injury is preconditioning. Preconditioning is caused by a weak damage of cells, which become more resistant to subsequent, more severe damage. We found that preconditioning by deprivation of trophic factors, or deprivation of trophic factor and glucose effectively protects neurons against subsequent toxic effects of glutamate. Deprivation of trophic factors plays a decisive role in the development of resistance, regardless of whether it has been combined with glucose deprivation or not. Neuronal protection is achieved when the deprivation lasts from 30 min to two hours and is kept for a period of from one to five days. Preconditioning is accompanied neuronal secretion of cathepsin B occurs. We suggest that this phenomenon is associated with a more general process of exocytosis of lysosomes triggered by deprivation of trophic factors.

Postejaculation maturation of spermatozoa (capacitation and acrosomal exocytosis) in bovine sperm was assessed using the method of staining with chlortetracycline and inhibitor analysis to identify the transition ways of calcium of intracellular depots under the influence of highly dispersed silica. It was shown that highly dispersed silica in a concentration of 0.001% stimulates capacitation but has no impact on the acrosome exocytosis in bovine sperm. Activated by highly dispersed silica, capacitation was inhibited in the presence of cytochalasin D and H-89 inhibitors, whereas nocodazole and Ro 31-8220 had no influence on this process. The joint action of theophylline and guanosine diphosphate stimulates an increase in the amount of capacitated sperm cells, similarly to highly dispersed silica; inhibitors cytochalasin D and H-89 restrict the capacitation of sperm activated by these compounds. At the same time, the joint effect of prolactin and guanosine triphosphate had no effect on the capacitation of spermatozoa; addition of nocodazole and Ro 31-8220 inhibitors did not alter the effect of prolactin and guanosine triphosphate on the capacitation of sperm. A hypothesis was put forward, according to which increase in the cryoresistance of spermatozoa of bulls under the influence of highly dispersed silica is, apparently, determined by the transition of Ca2+ between the intracellular stores in the direction from inositol triphosphate-sensitive to inositol triphosphate-insensitive intracellular stores of calcium. The obtained data allow us to expand the notion of the biochemical mechanisms of capacitation in the spermatozoa of bulls.

In experiments on frog (Rana ridibunda) neuromuscular junction the influence of cholesterol oxidation on the presynaptic vesicular cycle was investigated. Application of cholesterol oxidase (1 u. a.) during 1/2 hour led to the oxidation of - 0.007 mg cholesterol per 1 g tissue and reduced stability of lipid rafts in the nerve terminals. Using electrophysiological techniques it was shown that the cholesterol oxidation decreases the evoked neurotransmitter release. In experiments with fluorescent FM-dyes the depression of the synaptic vesicles exo-endocytosis and the dispersion of synaptic vesicles clusters were revealed. Comparative analysis of electrophysiological and optical data, as well as experiments with water soluble quencher of FM-dye indicated the possibility of some neurotransmitter release by "kiss-and-run" pathway, when short-lived fusion pore is formed. It was concluded that cholesterol oxidation inhibit evoked exocytosis, and also synaptic vesicle delivery from reserve pool to cites of exocytosis probably by break of the clusterization. Perhaps the synaptic vesicles of recycling pool release the neurotransmitter using the kiss-and-run mechanism.

The inhibitor of Rho-kinase Y-27632 induces non-secretory exocytosis in PC12 cells. The influence of this compound on central synapses remains uninvestigated. We showed that Y-27632 at the concentration 100 jtM led to spontaneous [14C]glutamate release in synaptosomes, which was not accompanied by plasma membrane depolarization. Membrane potential was registered by fluorescent dye DiSC3(5). Y27632 induced an increase of acridine orange fluorescence, exercising no influence over fluorescence of FM2-10 dye. These results suggest that Rho-kinase inhibition decreases pH gradient of synaptic vesicles not inducing exocytosis. Dissipation of the gradient leads to leakage of neurotransmitters to cytosol pumping them out by plasma membrane transporters. Our results show the involvement of Rho-dependent branch of intracellular signaling in regulation of pH gradient in synaptic vesicles.

Luminous spots with a diameter of 1-2 microm, which are clusters of "synaptic buds", were revealed in the muscular wall of the earthworm using endocytotic fluorescent dyes FM1-43, FM2-10 and FM4-64. Application of the membrane probe Dil that is capable of being subjected to anterograde axonal transport to abdominal ganglia of the nervous chain, and subsequent (in a day) staining of nerve formations by endocytotic dye FM4-64 showed complete imposition of the emission data of the dyes that fluoresce in different parts of the spectrum. Using fluorescent marker DiBAC4(3) showed an increased emission of neural elements with increasing concentration of K+ in the extracellular environment. Application of FM2-10 showed that the higher concentration of K+ in solution, and hence the depolarization of the nerve cells, the faster the upload of the dye, and vice versa, the process slowed down in the absence of K+ in the medium. The seizure and removal of FM2-10 were blocked in calcium-free solutions in the presence of Ca2+ buffers, BABTA or BABTA-AM, but only after a preliminary 40 min incubation. The processes of exo- and endocytosis occurred in the clusters of synaptic "buds" and were preserved in conditions of "rest". This vesicle cycle depends on membrane potential and concentration of K+ and Ca2+, and, it is very likely that the calcium sensor operates on the principle "all or nothing".

In experiments on the frog neuro-muscular preparations using electrophysiological (two electrode fixing of potential) and optical (fluorescent endocytic dye FM1-43) methods, the value of surface cholestertol for exo-endocytic cycle of synaptic vesicles at the prolonged rhythmic activity (20 Hz--3 minutes) was investigated. It is shown that extraction of cholesterol from surface membranes by methyl-betta-cyclodextrin (1 mM MCD) leads to the expressed shifts in recycling of synaptic vesicles. Exocytosis of vesicles is decreased, and oppression of processes leading to restoration of the number of vesicles of ready releasable pool is observed. Cholesterol replacement from external membranes and membranes of recycling synaptic vesicles in addition to above described effects breaks processes of endocytosis and recycle of synaptic vesicles. Thus, in the processes of exocytosis, the key role is played by cholesterol of plasmatic membranes, and endocytosis critically depends on the amount of cholesterol in the membranes of synaptic vesicles.

The influence of lanthanoids on exocytosis was investigated. It was shown that gadolinium increases the spontaneous release of the glutamate nonmetabolizing analogue [3H]D-aspartate. It was established using the fluorescent dye acridine orange that gadolinium and lanthanum induce exocytosis. The effect was dose-dependent and was maximum at 300 microM Gd3+. The exocytosis induced by gadolinium was calcium-independent. It is suggested that lanthanides induce a vesicular release of neurotransmitters by the mechanisms common for all polyvalent cations.

In experiments on the frog motor nerve endings of cutaneous pectoris muscle using fluorescent microscopy it has been shown that initiation of massive transmitter release of synaptic vesicles by high potassium solutions in using endocytotic marker FM 1-43 at the nerve terminals light spots occurred only at some of the nerve terminals or at the some parts of nerve terminal. It has been revealed that application of caffeine increased the number of light terminals. Using extracellular microelectrode recording, we showed that both high potassium solutions and caffeine increased frequency of miniature end-plate potentials in a dose-dependent manner. However, high potassium solutions always increased the frequency of spontaneous transmitter release while caffeine increased it only in some experiments. It was concluded that processes of exo- and endocytosis can be caused both by entry of Ca ions at the nerve ending during depolarization (high potassium solutions) and by Ca release from endoplasmic reticulum (caffeine). Possible spatial localization of endoplasmic reticulum at the motor nerve ending is discussed. The hypothesis of its role at the remodeling of synapse was proposed.

We compared the effects of adenosine and cholinergic agonist carbachol on spontaneous secretion during local application of K+, ionomycin, and sucrose increasing Ca2+ concentration in the nerve terminal. Adenosine and carbachol had no effect on Ca2+ entry, but modulated later stages of exocytosis.

Electrophysiological and optical methods were used to study exo- and endocytosis of synaptic vesicles underlying secretion of the neurotransmitter from motor nerve terminals in frog sternocutaneous muscle. Increase in extracellular concentration of K+ or sucrose produced similar increase in the frequency of miniature endplate currents recorded by extracellular microelectrode. Fluorescent microscopy revealed bright spots in nerve terminal during stimulation of secretion with high-potassium solutions in the presence of endocytosis marker FM1-43. These spots corresponded to clusters of synaptic vesicles that passed through the cycles of exo- and endocytosis. Subsequent high-potassium stimulation of exocytosis in normal Ringer solution led to disappearance of marker spots, while in hyperosmotic saline the spots were preserved. No spots were seen after stimulation of neurotransmitter secretion with sucrose in the presence of FM1-43. It is concluded that quantal secretion of the neurotransmitter in frog motor nerve endings can be realized via both complete exocytosis of synaptic vesicles with subsequent endocytosis and kiss-and-run mechanism with the formation of a temporary pore.

The effect of acetylcholine inducing the exocytosis of secretory vesicles, upon the development of changes in electrical resistance in mammal gland's alveoli, was studied in mice. A decrease in the ACh concentration and in potassium ions concentration gradient was followed by a deceleration of the reduction of transepithelial electrical resistance. The data obtained suggest that the exocytosis of secretory vesicles is not reflected in changes of transepithelial electrical resistance in mammal gland's alveoli.

The ultrastructure of hybridoma cells cultured with 5.10(-4) centrophenoxine (CP) has been studied. It is shown that CP effects hybridomas and prevents retrovirus exocytosis. The effect of CP on Ca-calmodulin system associated with cytoskeleton is suggested.

The functional activity of thrombocytes (aggregation, endo- and exocytosis) and erythrocytes (aggregation) in healthy persons given a course of interferon or reaferon treatment has been studied. The results obtained in this study indicate that these preparations produce a modulating effect on the functions of thrombocytes and erythrocytes of donors having shown abnormal functional activity of these blood cells prior to the course of treatment.

The aim of the present study is to elucidate the possible significance of metabolic control of the calcium permeability of synaptosomes induced by alpha-latrotoxin. The alpha-latrotoxin influence of processes of GABA release and reuptake is shown to depend on synaptosomal metabolism. It has been found that incubation by the synaptosomes during one hour at 37 degrees C or their treatment by 10(-3) M monoiodacetate during 10 min at 10 degrees C or at room temperature completely abolished or at least strongly decreased the ability of alpha-latrotoxin to induce the calcium influx. Under the same conditions the ability of alpha-latrotoxin to activate the GABA release and inhibit the GABA reuptake do not change. It is concluded that only formation of calcium channels by alpha-latrotoxin is strictly controlled by the level of synaptosomal metabolism.

The functioning of ganules of blood platelets participating in the reversible endocytosis reaction was investigated in rabbits. Eczocytosis was shown to be initiated by inducers of two types: proteolytic enzymes and compounds accumulated by the granules (biogenic amines, dyes, anesthetics and phenothiazines). Heparin inhibits thrombin-induced eczocytosis. During eczocytosis produced by accumulation of compounds by blood platelet granules, the cells lose their capacity for maintaining the aggregation state. The nature of blood platelet granules participating in the reversible endocytosis reaction is discussed.

The effect of subinhibitory concentrations of monomycin on the submicroscopic structures of 2 strains of S. typhi, 5 and 799 was studied. It was shown that formation of filamentoue forms, separation of the cell wall, thinning out of the cytoplasm granular component, increasing of the size of the matter with low electron optic density of fine granular structure were common in the cell response of both strains. Formation of vacuoli containing the thinned out granular component, filterable elements and complex membrane structures followed by their liberation into the medium and formation of the forms devoid of the cell walls was a characteristic peculiar property of the cell response in strain 799. The cells of strain 5 were characterized by formation of large granular osmiefilic matters and their excretion from the cells.

Mg-ATPase (1 mM) induces a decrease in the intensity of light scattering (I1) at 620 nm of rat brain synaptic vesicles (SV) suspended in sucrose, with this decrease being indicative of the swelling of the vesicles. The Mg-ATPase-induced swelling appears to be associated with the function of H+-ATPase of SV membranes, since it is completely abolished by the proton pump blocker dicyclohexylcarbodiimide and the protonophore carbonylcyanide m-chlorophenylhydrazone. The Mg-ATPase-induced swelling was enhanced in the presence of the permeable anion Cl- (in the range of 25-50 mM KCl). Ca2+ (and Mg2+) at high concentrations (0.1-1.0 mM) cause aggregation of the SV as measured by changes in the I1. Colchicine and cytochalasin do not affect SV swelling and aggregation whereas Mg-ATP (1 mM) lowers aggregation caused by Ca2+ (1 mM).

The conductivity of planar bilayer membrane comprising asolectin and phosphatidylserine (concentration ratio 9:1) in a buffer solution increased sharply in the presence of synaptic vesicles (SV) isolated from the rat brain and added to one side of the membrane only. The bilayer remained stable upon modification, and the conductivity increment was dependent on SV concentration in the range from 4 to 16 mu of the total protein per ml. If I mM CaCl2 was present in the buffer solution, the conductivity increased by 2 to 3 orders of magnitude upon the addition of SV at a final concentration of 3-4 mu protein per ml. The membrane was unstable and its rupture occurred often at an early stage of conductivity changes. In the absence of SV addition the membrane was stable, with its conductivity remaining unchanged for 2 h and more. With I mM CaCl2 addition to the solution already containing SV, no conductivity changes were observed, the cause perhaps, being Ca2+-induced SV aggregation.