Biomedical subjects

A L Zefirov

Publications and source records attributed to A L Zefirov.

At least 19 recordsLinked to original sources

PubMed2006

The role of extracellular calcium in exo- and endocytosis of synaptic vesicles at the frog motor nerve terminals.

In the present study we combined FM 1-43 imaging and electrophysiological recording of miniature end-plate currents (MEPCs) to determine the role of extracellular calcium in synaptic vesicle exo- and endocytosis at the frog motor nerve terminals. We replaced extracellular Ca2+ ions with other bivalent cations (Sr2+, Ba2+, Cd2+, Mg2+) or used a calcium-free solution and monitored fluorescent staining of the nerve terminals in the presence of caffeine, which promotes the release of Ca2+ from intracellular stores. Caffeine has induced FM1-43 internalization only in the presence of bivalent cations in the external solution. The exposure of the neuromuscular junction to caffeine in a calcium-free solution caused a reversible failure of FM 1-43 loading and an increase in the nerve terminal width. This effect of a calcium-free solution was not due to a decrease in exocytosis, because caffeine-induced FM1-43 unloading from the previously loaded nerve terminals, as well as a degree of the MEPCs frequency increase, was unchanged. We conclude that the presence of Ca2+ or other bivalent cations in extracellular space is necessary for endocytosis but not for exocytosis of synaptic vesicles, while transmitter release is promoted by efflux of Ca2+ from intracellular stores. The effect of extracellular Ca2+ on endocytosis might be driven by the non-specific interactions with membrane lipids.

Animals↗

PubMed2006

Mechanism of N-ethylmaleimide-induced contraction of the frog sartorius muscle.

We studied parameters of the frog sartorius muscle contraction initiated by ryanodine receptor agonists in the presence of ROS donors. We hypothesized that sodium nitroprusside and hydrogen peroxide inhibit initiation of contractions by N-ethylmaleimide and that this effect of ROS donors on parameters of N-ethylmaleimide-induced contractions is due to a direct effects of sodium nitroprusside and hydrogen peroxide on N-ethylmaleimide, but not to inactivation of ryanodine receptors in the sarcoplasmatic reticulum of frog skeletal muscle.

Animals↗

PubMed2006

Role of protein kinase C in the effect of ATP on contractile function of the isolated strip from mouse diaphragm.

We studied the effects of adenosine and ATP on contractile function of the isolated strip from mouse diaphragm. ATP significantly increased the strength of muscle contraction induced by carbachol. Adenosine had no effect on carbachol-induced muscle contraction. P2 receptor antagonist suramin abolished the effect of ATP. The positive chronotropic effect of ATP was not observed after treatment with specific protein kinase C inhibitor chelerythrine. Our results indicate that the effect of ATP on contractile function of mouse diaphragm is realized via protein kinase C.

Adenosine↗

PubMed2006

Effect of GABAergic and adrenergic agents on activity of Na+/K+ pump and Cl(-)-cotransport in somatic muscle cells of earthworm Lumbricus Terrestris.

GABA, baclofen, epinephrine, and norepinephrine hyperpolarized the membrane of earthworm somatic cells. This effect was prevented by furosemide, removal of Cl- from the medium, or activation of Na+/K+ pump by 3-fold increase external potassium concentration. It was hypothesized that GABA, baclofen, epinephrine, and norepinephrine stimulate Na+/K+ transport via specific receptor inputs, but their effect on resting potential can be realized only under conditions of working Cl- symport.

Adrenergic Agents↗

PubMed2006

The presynaptic effects of arachidonic acid and prostaglandin E2 at the frog neuromuscular junction.

Arachidonic acid and prostaglandin E2 decreased the frequency of miniature endplate potentials with producing any changes in the their amplitude-time parameters. Arachidonic acid and prostaglandin E2 decreased the quantum composition of endplate currents and the amplitude of the third phase of the nerve ending response, which reflects currents though potential-dependent K+ channels. A perineural method was used to demonstrate that arachidonic acid and prostaglandin E2 suppressed the nerve ending Ca2+ current. The cyclooxygenase blocker indomethacin increased neurotransmitter secretion and decreased the third phase of the nerve ending response. The effects of arachidonic acid and prostaglandin E2 on evoked neurotransmitter release were not seen in the presence of indomethacin, while the third phase of the response continued to show a reduction. It is suggested that prostaglandin E2 mediates the effects of arachidonic acid on spontaneous and evoked neurotransmitter secretion, Ca2+ currents, and Ca2+ -dependent K+ currents. In addition, arachidonic acid and prostaglandin E2 had their own effects on potential-dependent K+ currents in nerve endings.

Animals↗

PubMed2006

Effects of high-potassium solutions and caffeine on synaptic vesicle exoendocytosis processes in the frog neuromuscular junction.

Studies on frog skin-pectoris muscle preparations using vital fluorescent microscopy showed that stimulation of transmitter secretion using high-potassium solutions with the endocytosis marker FM 1-43 induced bright spots in all motor nerve terminals, these representing accumulations of vesicles undergoing the exoendocytic cycle in the active zones of nerve endings. Stimulation of transmitter secretion with caffeine evoked bright spots only in some nerve terminals and only in some parts of the terminals. In summer, the number of bright spots on stimulation of transmitter secretion by caffeine increased sharply. Extracellular recording of spontaneous synaptic signals showed that high-potassium solutions, like caffeine, produced dose-dependent increases in the frequency of miniature endplate currents. However, while high-potassium solutions always increased the frequency, this occurred with caffeine in only a proportion of experiments. This leads to the conclusion that exoendocytosis processes can occur both because of the influx of Ca(2+) ions into nerve endings as a result of depolarization (high-potassium solutions) and because of the release of Ca(2+) ions from the endoplasmic reticulum (caffeine). The possible spatial localization of the endoplasmic reticulum in nerve endings is discussed. The endoplasmic reticulum is suggested to have a role in synapse remodeling processes.

Animals↗

PubMed2006

[The intracellular calcium and mechanisms of endocytosis of synaptic vesicles at the frog nerve ending].

In the experiments on frog motor nerve endings of cutaneous pectoris muscle, made by extracellular recording of synaptic signals, it has been shown that the increase in intracellular calcium ion concentration in the nerve ending (by enhance of extracellular potassium ion concentration, or by addition of caffeine) leads to an increase in the miniature end-plate potential frequency, which is preserved over the whole period (about 10 min) of action of these substrates. The rhythmic stimulation of motor nerve (20 or 100 imp/s) quickly leads to a decrease in the end plate potentials amplitude. It has been shown by fluorescent microscopy with the use of endocytotic marker FM 1-43 that in the course of a short time exposition (5 min) in a high potassium solution (40 mM) or caffeine (5 mM), light spots appeared in the nerve ending. This shows that synaptic vesicles undergo intensive processes of endocytosis. During a longer exposition (30 min) no light spots were revealed, whereas the nerve ending width increased. This data allowed to propose that the process of endocytosis was blocked. In the presence of even lower concentrations of potassium ions and caffeine, and during a long rhythmic stimulation (20 or 100 imp/s) no blocking of endocytosis was revealed. It is concluded that high concentrations of intracellular calcium in the frog motor nerve ending leads to a reversible block of endocytosis, while exocytosis in synaptic vesicles is proceeding.

Animals↗

PubMed2005

Reactive oxygen species contribute to the presynaptic action of extracellular ATP at the frog neuromuscular junction.

During normal cell metabolism the production of intracellular ATP is associated with the generation of reactive oxygen species (ROS), which appear to be important signalling molecules. Both ATP and ROS can be released extracellularly by skeletal muscle during intense activity. Using voltage clamp recording combined with imaging and biochemical assay of ROS, we tested the hypothesis that at the neuromuscular junction extracellular ATP generates ROS to inhibit transmitter release from motor nerve endings. We found that ATP produced the presynaptic inhibitory action on multiquantal end-plate currents. The inhibitory action of ATP (but not that of adenosine) was significantly reduced by several antioxidants or extracellular catalase, which breaks down H2O2. Consistent with these data, the depressant effect of ATP was dramatically potentiated by the pro-oxidant Fe2+. Exogenous H2O2 reproduced the depressant effects of ATP and showed similar sensitivity to anti- and pro-oxidants. While NO also inhibited synaptic transmission, inhibitors of the NO-producing cascade did not prevent the depressant action of ATP. The ferrous oxidation in xylenol orange assay showed the increase of ROS production by ATP and 2-MeSADP but not by adenosine. Suramin, a non-selective antagonist of P2 receptors, and pertussis toxin prevented the action of ATP on ROS production. Likewise, imaging with the ROS-sensitive dye carboxy-2',7'-dichlorodihydrofluorescein revealed increased production of ROS in the muscle treated with ATP or ADP while UTP or adenosine had no effect. Thus, generation of ROS contributed to the ATP-mediated negative feedback mechanism controlling quantal secretion of ACh from the motor nerve endings.

Acetylcholine↗

PubMed2005

Mechanisms of hyperpolarizing effect of GABA on resting potential of the Lumbricus terrestris muscular wall somatic cells.

GABA, baclofen, isoguvacine increase, and cis-4-aminocrotonic acid does not modify resting membrane potential of muscle cells. Bicuculline, phaclofen, N-ethylmaleimide, chlorpromazine, verapamil, and removal of Ca2+ from bathing solution abolished the effect of baclofen, while U73122 and D609 were ineffective in this respect. The authors conclude that the Lumbricus terrestris muscle cells contain GABAergic structures similar to a- and b-receptors. Activation of GABA receptors induced Cl- inward current and Ca2+ entry with subsequent activation of calmodulin-like proteins, which causes membrane hyperpolarization by increasing the effect of "pumping potential" on resting membrane potential.

Animals↗

PubMed2005

The course of experimental myocardial infarction under conditions of Suppressed and enhanced NO synthesis.

Experimental myocardial infarction (coronary ligation without drug infusion) reduced heart rate, stroke volume, cardiac output, and blood pressure. Under conditions of blocked NO synthesis, experimental myocardial infarction was accompanied by a transient increase in heart rate, stroke volume, cardiac output, and blood pressure, followed by their decrease throughout the experiment. Experimental myocardial infarction against the background of L-arginine treatment increased all these parameters with their subsequent stabilization at the attained level. Animal survival after administration of L-arginine was 80% vs. 33 in the control, and 22% after administration of L-NAME. It can be hypothesized that L-arginine possesses an intrinsic NO-independent cardioprotective effect.

Animals↗

PubMed2005

[Mediators. Evolution of the concept].

The authors review the up-to-date situation in the issue of chemical mediators, acting in the nervous system. Mediators are key intercellular informational molecules found in the brain and peripheral nervous system. Most scientists interpret the term "mediator" rather widely, whereas others define it very strictly and clearly. That is why in neurophysiology the question of what a mediator is has not been answered fully yet. It is clear from the given scientific and historical review, that every new substance (after the first one, acetylcholine) that has had pretensions of being called a mediator, has altered the criteria of it. The authors present the contemporary classification of mediators, belonging to different classes of chemical compounds. Special attention is paid to chemical agents formed in the organism, which have only recently been referred to the class of mediators (gases and D-amino acids).

Animals↗

PubMed2005

[Presynaptic effects of arachidonic acid and prostaglandin E2 in the frog neuromuscular synapse].

Arachidonic acid and prostaglandin E2 decreased the frequency of miniature endplate currents without changing their amplitude-temporary parameters. They also reduced the evoked transmitter release and the amplitude of the 3rd phase of nerve ending response corresponding to the voltage-dependent K(+)-current. Using perineural recording, It was shown that arachidonic acid and prostaglandin E2 decreased the Ca2+ currents of nerve endings. Indometacin: inhibitor of cyclooxygenase, enhanced the evoked transmitter release and decreased the 3rd phase of nerve ending response. Indometacin prevented the effects of arachidonic acid on evoked transmitter release, whereas the effects of arachidonic acid on the 3rd phase was preserved. Prostaglandin E2 seems to mediate the effects of arachidonic acid on spontaneous and evoked transmitter release, Ca(2+)- and Ca(2+)-activated K(+)-currents. Moreover, the arachidonic acid and prostaglandin E2 exerted their own effects upon voltage-dependent potassium current of motor nerve ending.

Acetylcholine↗

PubMed2005

[Effects of high potassium solutions and caffeine on the processes of exo-endocytosis of synaptic vesicles at the frog motor nerve ending].

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.

Animals↗

PubMed2004

Kiss-and-run quantal secretion in frog nerve-muscle synapse.

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.

Animals↗

PubMed2004

Effect of blockers of potential-dependent and calcium-activated K+-channels on facilitation of neuromuscular transmission.

Rhythmic stimulation of nerve-muscle preparation of frog sternal muscle bathed in low-Ca(2+) saline increased the release of neurotransmitter (facilitation) and modified the shape of extracellular response of nerve terminal (decreased phase III amplitude). Iberiotoxin and 4-aminopyridine modified the dynamics these processes. We conclude that inactivation of potential-dependent K(+)-channels and activation of calcium-dependent K(+)-channels in frog motor nerve terminals during rhythmic activity modulate Ca(2+) influx into nerve terminals and contribute into facilitation of neurotransmitter secretion. The degree of these mechanisms depends on the rate of synaptic rhythmic activity.

4-Aminopyridine↗

PubMed2004

[The mechanisms of short-term forms of synaptic plasticity].

In experiments on the frog cutaneous pectoris muscle in cases of different external calcium concentrations, using extracellular recording technique, processes of facilitation and depression of transmitter release during the high-frequency stimulation were investigated. On the ground of experiments using intracellular mobile calcium buffers BAPTA-AM and EGTA-AM, it was proposed that at least two (low- and high-affinity) calcium-binding sites underlie the facilitation. Both the facilitation and the depression were accompanied by such transformations of underlied of nerve ending responses as changes of the third phase amplitude. Application of potassium channel blockers allowed us to reveal the significant contribution of changes of duration of the AP repolarisation phase and, accordingly, the changes of magnitude of calcium influx to development of facilitation and depression of transmitter release. It was also revealed that, during the high-frequency rhythmic stimulation, the increase of asynchrony of transmitter release leading to decrease of facilitation and increase of depression occurred. It was concluded that the forms of short-term synaptic plasticity--facilitation and depression, were caused by various presynaptic mechanisms: the increase of concentration of "local" and accumulation of "residual" calcium, the changes of calcium influx, increase of temporal course of secretion, the impairment of equilibrium between the depletion and restoration of mediator supply. Due to some of these processes and specific conditions of synapse functioning, the facilitation of the depression of transmitter release occurred.

Action Potentials↗

PubMed2003

Transmitter secretion in the frog neuromuscular synapse after prolonged exposure to calcium-free solutions.

Experiments on neuromuscular synapses from frog skin/chest muscle preparations in conditions of extracellular recording addressed changes in the spontaneous and evoked transmitter secretion after long-term (1.5-6 h) maintenance of preparations in calcium-free solution containing EGTA. Use of three microelectrodes for recording of single-quantum postsynaptic signals showed that calcium-free solution altered the characteristic topography of transmitter secretion in nerve terminals, with widening and fusion of groups of transmitter release. These changes persisted after preparations were returned to the initial solution. These data suggest that calcium-free solutions lead to disorganization of the active zones of nerve endings, At initially low extracellular Ca ion concentrations (0.15-0.4 mM), disorganization of active zones induced by prolonged maintenance of preparations in calcium-free solutions led to decreases in the mean amplitude of endplate currents (EPC) because of decreases in their quantum composition, increases in the time course of transmitter secretion, and decreases in the frequency of miniature endplate currents. The relationship between quantum composition of EPC and the extracellular Ca ion concentration showed a sharp displacement towards higher concentrations, without significant changes in the slope of the relationship. At high initial Ca concentrations (1.8 mM), long-term exposure to calcium-free solutions led to a less marked decrease in EPC amplitude. It is suggested that the extra- and intracellular Ca ion concentrations support the maintenance of the characteristic morphofunctional organization of the apparatus responsible for transmitter secretion in frog nerve endings. Disorganization of the active zones leads to disruption of elements involved in transmitter secretion and decreases in the efficiency of secretion.

Animals↗

PubMed2003

Effects of some transmitters on resting membrane potential of somatic cell in Lumbricus terrestris muscle wall.

Serotonin, glutamate, glycine, ATP, and muscarine had no effect on resting membrane potential of muscle cell in earthworm Lumbricus terrestris. Nicotine depolarizes and GABA hyperpolarizes the muscle membrane. Removal of K(+), Cl(-) and addition of ouabaine and strychnine to the solution abolished the effect of GABA. The authors conclude that the Lumbricus terrestris myocyte membrane contains nicotine receptors and GABAergic receptors sensitive to strychnine. Stimulation of these receptors activates sarcolemmal ionic pumps and causes membrane hyperpolarization.

Animals↗