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P A Doroshenko

Publications and source records attributed to P A Doroshenko.

16 recordsLinked to original sources
PubMed1992

Effects of serotonin and cAMP on calcium currents in different neurones of Helix pomatia.

Effects of application of serotonin (5-HT) and intracellular administration of cyclic adenosine monophosphate (cAMP) on voltage-gated calcium current (ICa) were studied in isolated, intracellularly perfused Helix pomatia neurones. Two types of the effects of 5-HT (1-10 microM) were observed in different neurones: reversible inhibition (by about 20%) or reversible potentiation (up to 50%) of the current amplitude. Some cells did not respond to 5-HT application. In cells with the potentiating effect of 5-HT, ICa could also be increased by intracellular introduction of cAMP (100 microM), but not the guanosine analogue, cGMP (50-100 microM). These effects were not additive and could be potentiated by theophylline (5 mM) and 3-isobutyl-1-methylxanthine (IBMX, 100-500 microM); they could be mimicked by forskolin (10-50 microM) and abolished by tolbutamide (1-5 mM) or protein kinase inhibitor (500 micrograms/ml), indicating that cAMP-dependent phosphorylation mediates the potentiating action of 5-HT on ICa. In neurones showing inhibitory effect of 5-HT, neither cAMP nor forskolin increased ICa. Methiothepin (10-50 microM), a 5-HT1,2 receptor antagonist, irreversibly inhibited the potentiating effect of 5-HT, while antagonists of 5-HT2 receptors cyproheptadine (10-50 microM) or ketanserine (10-60 microM) and of 5-HT3 receptors ISC 205-930 (10-50 microM) or cocaine (5-25 microM) had no effect on ICa and its enhancement by 5-HT. It is suggested that in certain snail neurones the possibility of cAMP-dependent up-regulation of ICa correlates with the presence of 5-HT1-like receptors in the neuronal membrane.

Animals↗
PubMed1990

[The effect of cAMP on the calcium currents of mollusk neurons possessing different sensitivities of their calcium conductance to serotonin action].

The action of cAMP (100 microM) and serotonin (5-HT, 1-10 microM) on the calcium current (Ica) in intracellularly perfused Helix pomatia neurons was studied with voltage clamp method. Three types of 5-HT-induced changes in the calcium current were observed in different cells: reversible blockade, increase and no changes in the current amplitude. Intracellular introduction of exogenous cAMP (100 microM) affected Ica only in cells with the stimulatory effect of 5-HT; cAMP-induced increase in the current amplitude was not additive to that elicited by 5-HT while both of these effects were similarly potentiated by cyclic nucleotide-phosphodiesterase inhibitor. The data presented show that the stimulatory action of 5-HT on the potential-activated calcium current is mediated by an increase in intracellular cAMP. Existence of two types of calcium channels differing in their dependence on cAMP metabolism is suggested in the snail neurons. The presence of the cAMP-dependent calcium channels seems to correlate with the existence of the definite type of 5-HT receptors in the cell membrane. A new approach to the investigation of isolated neurons is suggested: their functional identification.

Animals↗
PubMed1989

gamma-Aminobutyric acid elevates cytosolic Ca in bovine chromaffin cells.

Measurements of the cytosolic Ca concentration ([Ca]i) with the Ca-sensitive dye, fura-2, showed that in intact, but not in voltage-clamped, bovine chromaffin cells gamma-aminobutyric acid (GABA, 10 microM) elicited a transient increase in [Ca]i. The Ca transient of intact cells was inhibited by bicuculline (20 microM), by removal of extracellular Ca or by treatment with the Ca channel blocker cobalt (2.5 mM), and enhanced by lowering the extracellular Cl. We conclude, that GABA elevates [Ca]i by inducing a GABAA-receptor-linked Cl current which depolarizes the cell membrane sufficiently to activate potential-operated Ca channels and cause Ca entry into the cell.

Adrenal Medulla↗
PubMed1988

Modulation of calcium current by calmodulin antagonists.

The short-term effects of bath applied calmodulin antagonists--chlorpromazine, trifluoperazine and calmidazolium (R24571)--on potential-dependent calcium channels in the membrane of intracellularly perfused snail neurons were studied in voltage clamp conditions. All the drugs affected the calcium inward current peak value, the effects being reversible and dependent on the concentration used. Submicromolar concentrations (0.1-1 microM) increased the current amplitude (the maximal effect was on the average 20% at 0.5 microM), whereas higher concentrations inhibited the current. Analysis of the dose-effect curve for the blockade suggests positive cooperativity in the interaction of the drugs with the channel; experimental data on chlorpromazine action (10-100 microM) are well approximated by a binding curve for two molecules with the effective Kd = 70 microM. The efficiency of the blockade depended neither on the current-carrying cations (calcium or barium) nor on the intracellular introduction of 10 mM EGTA. The presence of calmodulin antagonists influenced the blockade of the calcium current by inorganic blockers: 50 microM chlorpromazine decreased the Kd value from 90 to 50 microM for the current blockade by Cd ions. It is suggested that calmodulin antagonists interact with two sites in the calcium channel, with high and low binding affinity (responsible for enhancement and inhibition of the current, respectively). The interaction induces changes in binding of penetrating cations in the channel, thereby producing modulation of the calcium current amplitude.

Animals↗
PubMed1988

[Action of forskolin on the calcium current in the membrane of nerve cells in mollusks].

The action of forskolin, adenylate cyclase activator, on the calcium inward current was studied on intracellularly perfused nonidentified snail neurons. Extracellular application of 5-20 mumol/l of forskolin was shown to cause about 20% increase of the calcium current amplitude and this confirms the previous suggestion that the stimulation of the cellular synthesis of cAMP leads to enhancement of the calcium current through the cell membrane.

Animals↗
PubMed1986

Transmembrane outward hydrogen current in intracellularly perfused neurones of the snail Helix pomatia.

The ionic nature and pharmacological properties of the outward current activated by membrane depolarization were studied on isolated neurones of the snail Helix pomatia, placed in Na+- and Ca2+-free extracellular solutions and intracellularly perfused with K+-free solution ("nonspecific outward current"). It was shown that the amplitude and reversal potential of this current (estimated from instantaneous current-voltage characteristics) are determined mainly by the transmembrane gradient for H+ ions. Lowering of pHi induced an increase in the current amplitude and a shift of the reversal potential to more negative values; the shift magnitude was comparable with that predicted for the hydrogen electrode. Raising pHi, as well as lowering pHo, induced a decrease in the current amplitude and a displacement of the current activation curve to more positive potentials. Addition of EGTA (8 mmol/l) to the intracellular perfusate did not affect the current amplitude. Extracellular 4-aminopyridine (10 mmol/l), verapamil (0.25 mmol/l) or Cd2+ (0.5 mmol/l) blocked the current. It is concluded that the current studied is carried mainly by H+ ions. In the same neurones the nature of the fast decay of the calcium inward current was also studied (in the presence of extracellular Ca2+ ions). This decay considerably slowed when pHi was raised or pHo was lowered, and it became less pronounced upon extracellular application of 4-aminopyridine or upon intracellular introduction of phenobarbital (4 mmol/l) and tolbutamide (3 mmol/l). It is suggested that the fast decay of the calcium inward current is due to activation of a Ca-sensitive component of the hydrogen current which depends on accumulation of Ca2+ ions. The possible physiological role of the transmembrane hydrogen currents is discussed.

4-Aminopyridine↗
PubMed1985

Fast decrease of the peak current carried by barium ions through calcium channels in the somatic membrane of mollusc neurons.

In experiments on nonidentified intracellularly perfused snail neurons the effects of replacing external divalent cations on the function of potential-dependent Ca channels have been studied. Ba substitution for Ca in the external medium caused a rapid decline (with half-times of about 2-3 min) in peak inward current amplitude when the current was activated from holding potential levels close to the resting potential. The decline could be reversed by membrane hyperpolarization. Barium current declined to a steady-state level which resembled in both relative amplitude (10-30% of the initial current amplitude) and insensitivity to intracellular introduction of exogenous cAMP the steady-state Ca current reached during the "wash out" process. It is suggested that two populations of Ca channels exist in snail neuronal membrane, one of which is dependent on cAMP metabolism and is reversibly switched off by the passage of Ba ions.

Animals↗
PubMed1979

[TEA-resistant outward current in the somatic membrane of perfused nerve cells].

The outward currents remaining after addition of 20-50 mM tetraethylammonium (TEA) to the extracellular solution were studied on perfused isolated neurons from Helix pomatia. A potassium-carried noninactivating outward current with potential-dependence and kinetics different from those of TEA-sensitive potassium currents was found. This TEA-resistant current includes a component depending on the presence of the inward calcium current. It could be abolished by replacing extracellular calcium by magnesium ions, by blocking the calcium channels with extracellular cadmium ions and their distruction by intracellular fluorid ions. An increase in the level of intracellular free carcium (by perfusing the cell with solutions containing Ca-EGTA buffer) potentiated the TEA-resistant component of the outward current and the removal of free calcium by EGTA decreased it. A conclusion is made that the somatic membrane contains outward current channels which can be activated only when calcium ions are bound to its inner surface.

Animals↗
PubMed1978

[Separation of potassium and calcium channels in the nerve cell soma membrane].

Calcium inward and potassium outward currents were studied on internally dialysed isolated neurons of the snail Helix pomatia. Different sensitivity of the corresponding channels to changes in external pH was found. This difference was used for separation of their activation regions on the potential axis so that the characteristics of the inward and outward currents could be studied with minimal overlap. It is shown that the outward current channels possess a definite permeability to Tris ions (PTris :PK=0.05). This explains the impossibility to switch off this current by substituting Tris for internal potassium. The channels for the inward calcium current inactivate slowly with a first order kinetic; their instantaneous current-voltage characteristic reveals considerable Goldman-type rectification. The selectivity of the calcium channels to other bivallent cations is Ba:Sr:Ca:Mg=2.8:2.6:1.0:0.2.

Animals↗
PubMed1978

[Study of the reversion potential for the slow component of the entering current in the membrane of mollusk neurons].

The nature of a significant deviation of the reversal potential for the slow inward current component from calcium equilibrium potential was investigated in the snail neuron somatic membrane. It is shown that this deviation may be due to a development of a non-specific outward current during large depolarizing shifts of the clamped membrane potential. The real equilibrium potential for calcium ions in nerve cells may be of about +200 mV.

Animals↗