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Biomedical subjects

S V Konev

Publications and source records attributed to S V Konev.

At least 37 records · Page 2Linked to original sources
PubMed1993

Kinetics and peculiarities of thermal inactivation of volume-induced Na+/H+ exchange, Na+,K+,2Cl- cotransport and K+,Cl- cotransport in rat erythrocytes.

The kinetics of the volume-dependent activation of Na+/H+ exchange, Na+,K+,2Cl(-)-cotransport and K+,Cl(-)-cotransport in rat erythrocytes was studied. The significant increase in the rate of Na+/H+ exchange is observed within 15 min after hypertonic shrinkage while the maximum transport rate is reached by 20 min. A delay of about 5 min was found in activation of Na+,K+,2Cl(-)-cotransport, the maximum transport rate being reached 10 min after shrinkage. Activation of K+,Cl(-)-cotransport by hypotonic swelling was registered within 10 min after cell swelling, with a simultaneous achievement of the constant transport rate. Preincubation of cells at 49 degrees C has no effect on the basal Na+/H+ exchange and Na+,K+,2Cl(-)-cotransport but suppresses the activation of these systems by osmotic shrinkage. On the contrary, the rate of K+,Cl(-)-cotransport in isosmotic medium is raised 10-fold after preincubation at 49 degrees C. The thermal treatment at 49 degrees C blocks the activation of K+,Cl(-)-cotransport by swelling. On the basis of the data on thermal denaturation of spectrin at the same temperature it was suggested that the cytoskeleton of erythrocyte membrane is involved in volume regulation of the ion-transporting systems and that the molecular mechanisms which underlie the activation of Na+/H+ exchange, Na+,K+,2Cl(-)-cotransport and K+,Cl(-)-cotransport are essentially different.

Animals↗
PubMed1993

Effect of gamma-irradiated carbohydrates on isolated synaptic membranes.

The effect of gamma-irradiated solutions of carbohydrates, mainly glucose, upon Na+, K(+)-ATPase and lipid peroxidation in rat brain synaptosomal membranes was studied. The membrane damage by irradiated glucose was enhanced in the presence of Fe2+ and was diminished when a free-radical scavenger (BHT) or metal chelators (EDTA, EGTA) were present. It is suggested that a key element in the free-radical membrane damage by irradiated carbohydrates is an Fe(2+)-complex of some species of the radiolysis products. Participation of radiotoxins of carbohydrate origin in radiobiological effects is discussed.

Animals↗
PubMed1992

Sensitivity of the brain synaptosomal membrane Mg(2+)-ATPase activity to arachidonic acid is under control of the Na+,K(+)-ATPase state.

Evidence is presented for the sensitivity of the synaptosomal plasma membrane Mg(2+)-ATPase activity to arachidonic acid being dependent on the functional state of Na+,K(+)-ATPase. An "Inversion effect" was observed at arachidonic acid concentrations exceeding 80 mumol/l when the Mg(2+)-ATPase activity (after ouabain addition) is higher than the total ATPase activity (without ouabain). The "Inversion effect" is reduced by cyclooxygenase inhibitor indomethacin or acetylsalicylic acid and restored by prostaglandin PGA2 or PGD2.

Animals↗
PubMed1991

On the mechanism of shrinkage-induced potassium influx in rat and human erythrocytes.

The rates of 86Rb influx into human and rat erythrocytes were studied in media of various tonicity. At sucrose concentrations below 0.3 mol/l, the ouabain-insensitive, furosemide-inhibited component of influx increased in rat but not in human erythrocytes; this may be explained by a rise in the rate of Na+, K+, Cl-- and/or K+, Cl-cotransport. An increase in osmolarity resulted in a reduction of this as well as of the ouabain and furosemide-insensitive component in rat erythrocytes. At the same conditions a drastic inhibition of Na+, K(+)-pump occurred both in rat and human erythrocytes. We failed to observe a lag-phase in the activation of the cotransport in rat erythrocytes; i. e. the process of activation parallels the shrinkage of cells. In rat erythrocyte ghosts, the shrinkage-induced stimulation of the cotransport was lost, and the direction of their osmotic reaction (inhibition of transport pathways) was similar to that in human erythrocyte ghosts. It is suggested that the mechanism of volume regulation of ion transport in intact cells involves a step of physical amplification via a change in interactions between the protein carcass and the lipid bilayer.

Animals↗
PubMed1991

[The coupling of Na,K-ATPase with the sodium channels in the plasma membranes of the brain synaptosomes of rats].

Effect of neurotoxins veratrine (100 micrograms/ml) and tetrodotoxin (1 microM) on the binding of 3H-ouabain (10(-8) M) with Na,K-ATPase of intact synaptosomes and isolated synaptic membranes was studied. The persistent opening of sodium channels in synaptosomes by veratrine results in an increase of specific binding of the labeled ligand by 20%. A similar effect was caused by Na/H exchanger monensin. Destruction of microtubules with vinblastine and colchicine has no influence on veratrine action, while depolymerization of microfilaments with cytochalasin B reverses the neurotoxin effect. In isolated synaptic membranes veratrine and tetrodotoxin stimulate ouabain binding, the absolute veratrine-induced increment being several times higher in the presence of ATP than in its absence. Since the closed vesicles of any type are not permeable to ATP and ouabain, it means that in the isolated membranes an interaction between sodium channels and Na,K-ATPase molecules takes place. In intact nerve endings such a mechanism may be operative along with the known ways of control of sodium pump and its ouabain-binding site.

Animals↗
PubMed1989

[Calcium transport in brain synaptosomes during depolarization. The role of potential-dependent channels and Na+/Ca2+ metabolism].

The contribution of Ca2+ channels and Na+/Ca2+ exchange to Ca2+ uptake in rat brain synaptosomes upon long- (t greater than or equal to 30 s) and short-term (t less than 30 s) depolarization by high K+ was studied by measuring the 45Ca content and free Ca2+ concentration (from Quin-2 fluorescence). At 37 degrees C, the system responsible for the K+-stimulated uptake of 45Ca (t greater than or equal to 30 s) and the Na+/Ca+ exchanger are characterized by a similar concentration dependence of external Ca2+ (Ca0(2+] and K0+ as well as by an equal sensitivity to verapamil (Ki = approximately 20-40 microM) and La2+ (Ki = approximately 50 microM). These data and the results from predepolarization suggest that the 45Ca entry into synaptosomes at t greater than or equal to 30 s is due to the activation of Na+/Ca+ exchange caused by its electrogenic component, while the insignificant contribution of Ca2+ channels can be accounted for by their inactivation. At low temperatures (2-4 degrees C) which decelerate the inactivation, the initial phase of 45Ca uptake is fully provided for by Ca2+ channels, showing a lower (as compared to the exchanger) affinity for Ca0(2+) (K0.5 greater than 1 mM)m a greater sensitivity to La3+ (Ki = approximately 0.2-0.3 microM) and verapamil (Ki = approximately 2-3 microM); these channels are fully inactivated by predepolarization with K0+, ouabain and batrachotoxin. The Ca2+ channels can be related to T-type channels, since they are not blocked by nicardipine and niphedipine.

Animals↗
PubMed1989

[Cytoskeletal regulation of the sensitivity of brain synaptosomes to the depolarizing action of veratrine].

Using a radioactive permeant cation 3H-tetraphenylphosphonium, the sensitivity of rat brain synaptosomes to depolarizing action of veratrine, which specifically opens the sodium channels, was compared before and after destruction of microtubules and microfilaments. Depolymerization of microtubules with colchicin and vinblastine decreased an apparent affinity of veratrine to its receptor in the channel, while destruction of microfilaments with cytochalasin B had the opposite effect. Colchicine did not change allosteric interactions between the receptor for veratrine and that for scorpion venom in the sodium channel evaluated by the ability of scorpion venom to facilitate veratrine-induced depolarization of synaptosomes. It is suggested that two main cytoskeleton subsystems control the state of sodium channels in the nerve ending.

Animals↗
PubMed1987

[The ATP depletion process of human erythrocytes studied by freeze fracturing].

By the freeze-fracture method it is shown that metabolic depletion of erythrocytes affects three levels of cell organization: the microstructural (erythrocyte form), microstructural (micro-relief of erythrocyte surface) and ultrastructural (ultrastructural state of erythrocyte plasma membranes) ones. As it is established, the size of spikes on the echinocyte surface and that of membrane vesicles budding from a cell coincide with each other. The structural modification of the membrane precedes the stage of erythrocyte crenation. The following model of vesicle budding process is suggested: reduction of ATP level and dephosphorylation of actin-spectrin network--structural modification of the protein and lipid membrane phases with the formation of regions disconnected from the spectrin framework--protrusion of these anomalous regions in the form of spikes--budding of spikes as spherical vesicles.

Adenosine Triphosphate↗
PubMed1986

[Characteristics of the heterogeneity of the physical properties of the lipid phase of synaptic membranes from the rat brain using the fluorescent probe pyrene].

A theoretical analysis of the dependence of pyrene excimerization on its concentration in biological membranes was carried out. It was shown that in synaptic membranes the concentration dependencies of pyrene excimerization parameter upon direct stimulation appear as nonlinear, thus being reflective of the heterogeneity of physical properties of the lipid phase. Upon pyrene excitation at the expense of the energy transfer from tryptophanyl residues the dependence is linear, which points to the homogeneity of the anular lipid pool. It was assumed that a comparison of microviscosity of the anular and bilayer lipids requires independent measurements of the coefficient of the probe distribution between the lipids or of the anular lipid content in the membrane.

Animals↗
PubMed1986

Inactivation of muscarinic acetylcholine receptors in brain synaptic membranes by free fatty acids. Evaluation of the role of lipid phase.

Arachidonic, linolenic and linoleic acids decreased the binding of the m-cholinergic antagonist [3H] QNB and did not affect the ratio of high to low affinity binding sites to the agonist carbamoylcholine in rat brain synaptic membranes. In the presence of arachidonic acid, SH-reagent N-ethylmaleimide acquired the ability to block QNB binding to receptor. Lipids in the bilayer and annular regions were probed by fluorescence of 1,6-diphenyl-1, 3, 5-hexatriene and pyrene. A microviscosity drop induced by increasing temperature from 10 to 37 degrees C did not affect the level of QNB equilibrium binding, whereas arachidonic acid strongly inhibited the binding at concentrations inducing the same drop in microviscosity as that induced by heating. For various unsaturated fatty acids an equal extent of receptor blocking was reached at quite different degrees of bilayer fluidization, the state of annular lipid being not changed under these conditions. It is suggested that the effect of unsaturated acids is reached through their direct interaction with the receptor, which undergoes a conformational change, rather than by an alteration of the physical state of the lipid phase of the membrane.

Animals↗
PubMed1986

[Ultrastructural changes in the erythrocyte plasma membranes of chickens in the early stages of interaction with the influenza virus].

Using freeze-fracture method it is shown that at the early stages of interaction with chicken erythrocytes the influenza virus induces generalized changes in the physical state of erythrocyte plasma membranes resulted from local disturbances of the cell surface in adsorption sites of virions. Ultrastructural changes seen in the membranes reach their maximum 5 minutes after infection which corresponds to the moment of maximum pinocytotic cell activity in relation to the viruses.

Animals↗
PubMed1986

[Intercellular interactions as a reason for the influence of the erythrocyte concentration on the parameters of detergent hemolysis].

Possible reasons are discussed for the abnormal erythrocyte concentration influence on the course of detergent hemolysis curves. The data obtained lead to a conclusion that erythrocyte plasma membrane properties and, as a consequence, the parameters of erythrocyte-detergent interaction are dependent on cell concentration. The dependence is more sharply expressed for concentrations 5.10(7) cell/ml. In the concentration range (5-20) X 10(7) cell/ml the state of erythrocyte membrane remains apparently unchanged.

Cell Communication↗
PubMed1984

[Changes in the orientation of the fracture plane of cell membranes during freeze-fracturing as an index of change in membrane structure].

Using freeze-fracture method it is shown that the direction of the fracture plane is a sensitive indicator of the membrane structural organization. A decrease in the proportion of longitudinal fractures is noted upon the membrane rearrangement presumably associated with changes in the lipid phase, and, on the contrary, an increase in the proportion of these fractures is observed upon some actions resulting, presumably, in changes of the protein phase of membranes.

Candida↗
PubMed1983

Effect of tetracaine on veratrine-mediated influx of sodium into rat brain synaptosomes.

Depolarization of synaptosomes with veratrine (0.1 mg/ml) or 50 mmol/l of KCl results in the release of radioactivity from 14C-choline loaded synaptosomes with partial dependence on external Ca2+. Like tetrodotoxin, tetracaine prevented veratrine but not KCl action, with a half-maximal effect at approximately 10(-5) mol/l of anesthetic. A similar half-maximal value was obtained for tetracaine blockade of veratrine-stimulated tetrodotoxin-sensitive 22Na influx into synaptosomes, complete blockage being achieved at 10(-4) mol/l. At this concentration tetracaine failed to modify Ca2+ channels measured by KCl-induced 45Ca uptake. Microviscosity of a lipid bilayer in synaptic membranes evaluated with 5- and 16-doxylstearate spin labels decreased at tetracaine concentrations exceeding 10(-3) mol/l. It is suggested that Na+ channels of synaptosomes are blocked by direct action of anesthetic or through changes in the channel annular lipids.

Acetylcholine↗
PubMed1981

[Sites responsible for stimulation of brain Na,K-ATPase by serotonin].

Serotonin (10(-6)-10(-3) M) stimulates Na,K-ATPase in the rat brain cortex homogenate with a maximal effect at 10(-4) M. Deseril (10(-5) M), antagonist of serotonin receptors, removes the stimulating effect. Deseril has no influence on noradrenaline-induced ATPase activation; the alpha-adrenergic blocker phentolamine does not affect serotonin activation. The effects of the two transmitters are additive. It is assumed that the mediators interact with different membrane sites, serotonin activation being initiated via the serotonin receptors.

Animals↗