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Vol 11, No 4 (2017)

Reviews

The influence of glucocorticoids and catecholamines on the neuromuscular transmission

Grishin S.N., Gabdrakhmanov A.I., Khairullin A.E., Ziganshin A.U.

Abstract

The review surveys the impact of “stress hormones”–glucocorticoids and catecholamines–on the functioning of the neuromuscular synapse. The review brings together the data on the influence of the main agents of stress–cortisol and norepinephrine–on the intensity and timing of the acetylcholine release, as well as signaling effect of its co-mediator ATP.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):253-260
pages 253-260 views

Articles

Inhibition of spontaneous synchronous activity of hippocampal neurons by excitation of GABAergic neurons

Zinchenko V.P., Gaidin S.G., Teplov I.Y., Kosenkov A.M.

Abstract

The molecular mechanisms of the neuronal spontaneous synchronous activity (SSA) regulation by population of GABAergic neurons have been investigated in rat hippocampal culture. The neurons from this population contain Ca2+-permeable KA receptors on the presynaptic membrane. Using image analysis, confocal microscopy and immunocytochemistry, we identified by the shape of Ca2+ signal the population of GABAergic neurons with unique charachteristics allowing these neurons to control SSA. The SSA in a neuronal network was suppressed by the KA-receptor mediated [Ca2+]i increase in neurons of this population. Agonists of GluR5/GluK1-containing KA receptors (domoic acid (DA), SYM2081, and ATPA) evoked a fast high-amplitude Ca2+ signal without desensitization only in this population of neurons. This fact points to Ca2+ permeability of KA receptors in these neurons. The GABA(A) receptor antagonist bicuculline increased the activity of AMPA but not KA receptors of these neurons, indicating presynaptical localization of KA receptors. Depolarization of cells induced by KCl (unlike bicuculline-induced depolarization) increased the activity of AMPA and KA receptors twofold, which points to the dependence of the activity on depolarization. A tenfold increase of the SSA frequency in neurons of this population caused an increase in the basal [Ca2+]i level, which was accompanied by inhibition of SSA in another numerous population of neurons, suggesting that an increased GABAergic inhibition takes place. Prolonged high-frequency oscillations causes a global [Ca2+]i increase in the neurons of this population and their subsequent death. Thus, KA receptors in the population of fast GABAergic neurons may implement a negative feedback under hyperexcitation by glutamate enhancing GABA release due to the fast and prolonged [Ca2+]i increase. It has been shown that this mechanism can be used to suppress hyperactivation of a certain population of neurons under high-frequency SSA and ischemia. It is obvious that selective death of inhibitory neurons from this population may lead to hyperexcitability of certain brain regions.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):261-274
pages 261-274 views

Kinetic characteristics of acetylcholinesterase and structural-functional state of rat erythrocyte membranes at moderate hypothermia

Klichkhanov N.K., Dzhafarova A.M., Al-Rabeei M.A.

Abstract

Artificial hypothermic state of homeothermic animals contributes to the stimulation of free radical processes in red blood cells. In order to understand what are the consequences of oxidative damage of erythrocyte membrane, we examined the dependence of the kinetic characteristics of integral membrane enzyme acetylcholinesterase (AChE) and structural and functional state of the membrane on the duration of mild hypothermia. For this purpose we reduced body temperature of adult Wistar rats by external cooling to 30°C (short-term moderate hypothermia) and then prolonged hypothermia up to 1.5 and 3 h. A short-term hypothermia was followed with an increase in Vmax and a decrease in Km, promoting an increase in the catalysis effectiveness.The optimum point on the graph of the concentration dependence was shifted to the area of lower concentrations, and the character of enzyme–substrate interactions at high concentrations of the enzyme changed. Upon prolongation of hypothermia, changes in the AChE kinetic characteristics favored normalization of the enzyme activity and concentration dependence. To test the hypothesis of a possible influence of the lipid matrix on the kinetic characteristics of AchE, we studied structural properties of the erythrocyte membranes using fluorescent probe pyrene. The observed changes in the structural and dynamic characteristics of erythrocyte membranes after a 1.5-h hypothermia suggested a reduction in microviscosity of both total and annular lipids. Prolongation of hypothermia up to 3 h favored normalization of this parameter. It was found that the indicators of the structural state of erythrocyte membranes at different durations of hypothermia correlate with certain kinetic characteristics of AChE. The data indicate that the prolongation of mild hypothermia up to 3 h triggers adaptive mechanisms directed to normalization of the erythrocytes membrane functioning.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):275-286
pages 275-286 views

Fatty acid composition of total lipids from needles and cultured calluses of conifers Pinus sylvestris L., Picea pungens Engelm., Pinus koraiensis Siebold & Zucc., and Larix sibirica Ledeb.

Semenova N.V., Makarenko S.P., Shmakov V.N., Konstantinov Y.M., Dudareva L.V.

Abstract

A comparative analysis of the fatty acid (FA) composition of lipids in the calluses and needles of Scots pine (Pinus sylvestris), white spruce (Picea pungens), Korean pine (Pinus koraiensis), and Siberian larch (Larix sibirica) has been carried out using the method of gas-liquid chromatography–mass spectrometry. In all cases, the total content of monounsaturated FA in the lipids of callus was higher than in the lipids of needles. Oleic acid (C18:1) represented a significant part of the composition of these acids. It was established that the lipids of callus of the studied species are characterized by a lower relative content and a smaller variety of Δ5-acids than the lipids of needles of the same species. It was shown that species-specificity of FA composition of the lipids of needles remains the same in the plant tissues grown in culture in vitro and does not depend on the material used to initiate the callus. Qualitative differences in the FA composition of the callus in culture and the needles of whole plant were detected. It was shown that the embryogenetic ability of the callus is associated with high content of oleic acid.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):287-295
pages 287-295 views

Identification of sterol-containing domains in vacuolar membranes by confocal microscopy

Nurminsky V.N., Nesterkina I.S., Spiridonova E.V., Rakevich A.L., Ozolina N.V.

Abstract

Sterol-containing domains in the tonoplast of beet root cells were identified by confocal microscopy with a membrane probe filipin. The presence of sterol-containing domains in the tonoplast was confirmed using methyl-β-cyclodextrin. Sterol-containing domains with separate small intensely fluorescing sterol-enriched sites were found in the vacuolar membrane; these sites can be attributed to raft structures. It was shown that sterol content in the isolated rafts can be assessed by confocal microscopy. β-Sitosterol, stigmasterol, α-tocopherol, and cholesterol were identified among the sterols of the isolated rafts.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):296-300
pages 296-300 views

The effect of dehydroepiandrosterone on inflammatory response of astroglial cells

Buyanova S.M., Chistyakov D.V., Astakhova A.A., Sergeeva M.G.

Abstract

An increased interest in neuroinflammation is conditioned by its involvement in various pathological processes in the brain. Astrocytes play an important role in neuroinflammation, participating in its regulation, throwing out a large number of signaling molecules. Steroid compounds, actively produced by astrocytes, are of interest with regards to the regulation of inflammatory processes in the central nervous system. In the present work the effect of dehydroepiandrosterone (DHEA) on astroglial cells (cultured primary rat astrocytes) in a model of inflammation was studied. The inflammatory response was stimulated with lipopolysaccharide (LPS). Expression levels of pro-inflammatory factor TNFα, antinflammatory interleukin IL-10, and both pro- and antiinflammatory protein COX-2 were measured. The expression of IL-10, COX-2, and TNFα mRNA was determined by real-time PCR, COX-2 protein level by immunoblotting method, TNFα and IL-10 release by enzyme immunoassay. The effect of short-term (30 min) and long-term (24 h) exposure to DHEA was evaluated. It was shown that DHEA potentiates LPS-stimulated (1) increase in the IL-10 mRNA level; (2) IL-10 release; (3) does not affect TNFα level, and (4) exerts a weak pulsating bidirectional effect on COX-2. Using trilostane, an inhibitor of 3β-hydroxysteroid dehydrogenase, a key enzyme of DHEA metabolism, it was shown that DHEA metabolites make the main contribution to its effect. Thus, DHEA is of interest as a stimulant of anti-inflammatory processes in the brain.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):304-310
pages 304-310 views

The role of pannexin 1 in the purinergic regulation of synaptic transmission in mouse motor synapses

Miteva A.S., Gaydukov A.E., Shestopalov V.I., Balezina O.P.

Abstract

The role of pannexin 1 in the release to the extracellular space of ATP/adenosine modulating the acetylcholine (ACh) secretion was studied in mouse diaphragm motor synapses. Using neuromuscular preparations obtained from wild-type and pannexin-1 knockout mice, the miniature endplate potential (MEPPs) and evoked endplate potentials (EPPs) were recorded in combination with pharmacological modulation of P2-type ATP receptors and A1-type adenosine receptors. Selective inhibition of A1 receptors with DPCPX or P2 receptors with PPADS increased quantal content of EPPs in wild-type mice. MRS 2211, selective antagonist of P2Y13 receptors, produced the same effect. Activation of receptors A1 or P2Y13 by their agonists (2-CADO and IDP, respectively) decreased the EPP quantal content. It means that the activity of endogenous ATP and adenosine is synergistic and directed to depression of the ACh release. ARL67156, an inhibitor of synaptic ecto-ATPases, which blocks the hydrolysis of ATP to adenosine and increases the level of ATP in the synaptic cleft, prolonged EPPs without changing their quantal content. In pannexin-1 knockout mice there were no changes in the EPP quantal content and in other parameters of synaptic transmission as compared to wildtype mice. However, downregulation of purinergic effects with antagonists of A1 or P2 receptors (DPCPX, PPADS, MRS 2211) did not change EPP quantal content and any other parameters of spontaneous or evoked ACh release in all cases. ARL67156 did not alter the temporal parameters of EPPs, either. Nevertheless, 2-CADO, the A1-type receptor agonist, decreased the EPP quantal content, while the agonist of P2Y13 receptors decreased the MEPP amplitude. Thus, in mice lacking pannexin 1, procedures revealing the presence and regulatory activity of synaptic ATP/adenosine did not change the parameters of synaptic transmission. The obtained data substantiate a mandatory role of pannexin 1 in the purinergic regulation of motor synapse activity by endogenous ATP/adenosine.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):311-320
pages 311-320 views

On the mechanism and functional significance of the ADP/ATP carrier (AAC) dimerization

Moiseeva V.S., Murugova T.N., Vangeli I.M., Byvshev I.M., Ravaud S., Simonyan R.A., Gordeliy V.I., Pebay-Peyroula E., Yaguzhinsky L.S.

Abstract

According to previous studies, ADP/ATP carrier (AAC) can possibly exist as a monomer or in a dimer state in the inner mitochondrial membrane; however, the question on its functional oligomeric state is still open. The aim of the present work is to establish the external factors that could control the functional oligomeric state of AAC (i.e., monomer or dimer). The study is based on the results of our previous work, which revealed that the volume regulation system of mitochondria (MVRS) affects the oxidative phosphorylation (OXPHOS) system: MVRS could transfer OXPHOS system functioning in a state of supercomplex. Consequently, one may expect that the volume regulation system could also control the functional state of AAC during phosphorylation. Here, on rat liver mitochondria we show that, depending on the incubation medium tonicity, AAC functions in two different ways: either as a monomer (in hypotonic and isotonic media) or as a dimer (in a hypertonic medium). Thus, the transition between the monomeric and dimeric forms of AAC is regulated by MVRS, as well as by functioning of OXPHOS. We conclude that the structural reorganization of AAC is associated with the entire OXPHOS reorganization into a supercomplex. It was also found that dimerization of AAC can occur not only due to the action of MVRS (in hypotonic media) but also under hypoxic conditions.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):321-329
pages 321-329 views

Sip1 mutation suppresses the resistance of cerebral cortex neurons to hypoxia through the disturbance of mechanisms of hypoxic preconditioning

Turovsky E.A., Babaev A.A., Tarabykin V.S., Turovskaya M.V.

Abstract

Mutation of Sip1 plays a key role in pathogenesis of the Mowat–Wilson syndrome characterized by the presence of pronounced epileptic signs in patients. As a rule, neurodegenerative processes are accompanied by hypoxic phenomena, glutamate toxicity, and death of nerve cells. The molecular mechanisms underlying these phenomena are multifaceted and complex. Hypoxia causes the leakage of glutamate and other neurotransmitters and thus activates glutamate receptors and channels of plasma membrane. Hypoxia is accompanied by increased synthesis and secretion of proteins-regulators of oxidative stress, inflammation, apoptosis, and synaptic transmission. In this work, we investigated the effect of Sip1 mutations on the neuronal sensitivity of the brain to hypoxia and the formation of the hypoxic preconditioning phenomenon. The preconditioning effect was evaluated by the degree of suppressing activity of NMDA receptors by hypoxic episodes. Using fluorescent microscopy, we showed that cortical neurons from the brain of heterozygous (Sip1wt/fI) and homozygous (Sip1fI/fI) mice are characterized by the absence of the hypoxic preconditioning effect, whereas in Sip1wt/wt neurons the amplitudes of Ca2+ responses to the application of NMDA are suppressed after transient episodes of hypoxia. In addition, hypoxia exerted a significant toxic effect on Sip1fI/fI neurons, which was manifested not only by an increased sensitivity to a decrease of the oxygen partial pressure (pO2) and increased amplitudes of Ca2+ responses to application of NMDA after each hypoxic episode, but also by death of a considerable number of cells.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):330-337
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Short Communication

Brassinosteroid-induced accumulation of complex oxylipins in flax leaves

Fedina E.O., Yarin A.Y., Blufard A.S., Chechetkin I.R.

Abstract

The effect of the steroid plant hormone 24-epibrassinolide on the content and composition of complex oxylipins in plants was studied for the first time, using flax as a model plant. In plants treated with the plant hormone, as well as in plants inoculated with Pectobacterium atrosepticum, the content of linolipins (galactolipids containing divinyl ether residues) increased. Hormonal pretreatment of the plants with subsequent infection resulted in a more significant accumulation of linolipins B (21-fold) and A (2.8-fold) as compared to the untreated (control) plants. The data suggest that brassinosteroids are involved in the regulation of the formation of complex oxylipins in pathogenesis of plants.

Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2017;11(4):301-303
pages 301-303 views

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