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Biomedical subjects

S V Konev

Publications and source records attributed to S V Konev.

At least 19 recordsLinked to original sources

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

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

[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

[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

[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

[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

[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

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

[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

[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

[Temperature-dependent structural rearrangements in the cytoplasmic membranes of Escherichia coli cells].

By the freeze-etching method, in has been shown that E. coli plasma membranes undergo a structural transition in the range of temperatures within 0 and 20 degrees C which could be observed as redistribution of intramembrane particles with free-zone formation. The onset of temperature interval of this transition (20 degrees) well correlate with the break in the Arrhenius curves characterizing the cell membrane permeability for free nucleotides and for respiration intensity.

Cell Membrane

[Mechanism of brain Na,K-ATPase activation by adrenaline].

Norepinephrine stimulates Na, K-ATPase from rat brain homogenates at concentrations of 10(-4)--10(-5) and 10(-7)--10(-8) M. A low concentration maximum is observed after 48 hrs of incubation at -20 degrees C and is not changed by the addition of alpha-tocopherol, glycerol and MAO inhibitor ipraside. The maximum observed at the mediator concentration equal to 10(-4)--10(-5) M is eliminated after treatment with EGTA. At all concentrations of norepinephrine the enzyme stimulation is removed by the alpha-adrenoblocker phentolamine. The activated enzyme reveals lower sensitivity to Ca2+ induced inhibition. The role of Ca2+ and conformational state of the membranes in the realization of the remote effect on the adrenoreceptor-Na, K-ATPase system is discussed.

Animals

[Effect of cAMP on the structural and functional properties of the erythrocytes].

Cyclic AMP inhibits the anion transport and decreases the osmotic resistance and deformability of erythrocytes with the normal level of ATP. With ATP-depleted erythrocytes cAMP exerted the opposite effects on the corresponding characteristics. In addition, it was observed that the pattern of cAMP effect on the cell form depends on the basal level of ATP. These effects may be associated with the two types of structural rearrangements of erythrocyte membranes established earlier: a cooperative transition not connected with protein kinase system, and a non-cooperative one caused by protein phosphorilation.

Biological Transport, Active

[Effect of the critical fragmentation of erythrocyte membranes].

After sonification of erythrocyte membranes, some changes were registered in these including a loss of their ability to structural rearrangements caused by cAMP (ESR-spectroscopy and luminescence data), an increase in cAMP binding and aggregation of intramembrane particles (freeze-fracture data). These findings suggest a non-identity of the structural organization in membranes and in their fragments. The cooperative nature of membrane structural modification at ultrasonic fragmentation is shown.

Cell Fractionation