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O Krishtal

Publications and source records attributed to O Krishtal.

At least 19 recordsLinked to original sources
PubMed2006

The agonists for nociceptors are ubiquitous, but the modulators are specific: P2X receptors in the sensory neurons are modulated by cannabinoids.

P2X2 and P2X3 receptors expressed in mammalian sensory neurons participate in nociception. Cannabinoid receptors modulate nociceptive processing in various models of pain. They are also expressed in nociceptive sensory neurons. We have examined the effect of cannabinoids on the slow P2X2 and P2X2/3 receptors in the cells isolated from nodosal and dorsal root ganglia of rat. The study was carried out by means of the whole-cell patch clamp and rapid superfusion methods. We have found that both endogenous and synthetic cannabinoids (anandamide, WIN55,212-2, and (R)-(+)-methanandamide) inhibit the slow response to ATP mediated by P2X2 and P2X2/3 receptors in a majority of tested neurons. This inhibition was significant but only partial: anandamide (0.5-1 microM) inhibited the response to 51+/-21% of control. In the remaining minority of tested neurons, the response was transiently facilitated. The effect of cannabinoids appears to be mediated via cannabinoid CB(1) receptors: it was reversibly inhibited by selective CB(1) antagonist, SR141716A (10 microM). Introduction of cyclic AMP (0.5 mM) into the cell potently facilitated the inhibitory effect of cannabinoids: the ATP-activated current was inhibited to 13+/-10% of control. These data indicate that cannabinoids may inhibit nociceptive responses produced by P2X receptors.

Adenosine Triphosphate↗
PubMed2005

Opioids inhibit purinergic nociceptors in the sensory neurons and fibres of rat via a G protein-dependent mechanism.

We have found that opioid and P2X receptors are functionally coupled in the sensory nerve fibres and neurons of rat. When examined in the skin-nerve preparation, the ATP-evoked discharges of nerve fibres belonging to n. saphenous were inhibited by various opiates in a naloxone-dependent manner. The functional coupling between opioid and purinergic receptors was studied in the neuronal cell bodies isolated from dorsal root and nodose ganglia. Both fast (mediated by P2X(3) receptors) and slow (P2X(2/3) heteromeric receptors) responses of sensory neurons to ATP were inhibited by opioids. The inhibition of slow responses developed in a characteristic biphasic manner: an initial short phase of potentiation (lasting for 300-400 s) was followed by long-lasting inhibition of the response (for about 50% when saturated). Both phases of the response were initiated by the application of the highly selective ligand for mu-receptors, endomorphin 1 (30 nM). Intracellular GTPgammaS caused a partial inhibition of the ATP responses and opioids were not effective against the residual response. Intracellular GDP eliminated the effects of opioids, while pertussis toxin (PTX) abolished only the inhibition phase. Thus, P2X receptors in the sensory neurons are affected by opioids via multiple G protein-dependent pathways.

Action Potentials↗
PubMed2004

Modulatory action of RFamide-related peptides on acid-sensing ionic channels is pH dependent: the role of arginine.

Acid-sensing ionic channels (ASICs) are involved in such functions of the sensory nervous system as mechanoreception, nociception and perception of acid taste. Phe-Met-Arg-Phe amide-related (FMRFa-related) peptides in micro m concentrations slow down the rate of ASICs desensitization. Here we report that this effect is strongly pH dependent: the lower the pH used to activate ASICs, the larger is the modulatory effect of Arg-Phe amide-related (RFa-related) peptides. Pre-application of the peptides results in a change to the desensitization kinetics of the ASICs-operated current from monoexponential to biexponential: the fast component retains the control kinetics, whereas the slow one is induced by the peptide. The lower the pH, the larger is the slow component, whereas there is practically no modulation at pH 6.6. Phe-Met-Val-Phe amide (FMVFa), which has neutral valine instead of arginine, similarly modulates the kinetics of ASICs, but does not reveal pH dependence of this action. Thus, positively charged arginine regulates the access of the RFa-related peptides to the modulatory site. We suggest that the pH dependence of the modulatory action of RFa-related peptides can be associated with the interaction of a positively charged arginine with histidine residues in the molecule of ASIC.

Acid Sensing Ion Channels↗
PubMed2003

P2X receptor-mediated excitatory synaptic currents in somatosensory cortex.

Fast P2X receptor-mediated excitatory postsynaptic current (EPSC) was found in pyramidal neurones of layer V of somatosensory cortex in slices acutely isolated from the brain of 17- to 22-day-old rats. The EPSCs were elicited by field electrical stimulation in the layer VI at 0.1 Hz in the presence of picrotoxin. When the glutamatergic EPSC was blocked by glutamate receptors inhibitors NBQX and D-AP5, a residual EPSC (rEPSC) was recorded from 85% of neurones tested. This rEPSC was not affected by blockers of nicotinic (hexamethonium) and serotonin (Y25130) receptors; however, it was reversibly inhibited by P2X receptors antagonists (NF023, NF279, and PPADS). An application of ATP (20 microM), beta,gamma-methylene ATP (25 microM), and alpha,beta-methylene ATP (20 microM) to acutely isolated pyramidal neurones of layer V evoked inward currents (30 to 400 pA) in 75% of cells tested. We concluded that several subtypes of P2X purinoreceptors participate in synaptic transmission in neocortex.

Adenosine Triphosphate↗
PubMed2002

Ionotropic P2X purinoreceptors mediate synaptic transmission in rat pyramidal neurones of layer II/III of somato-sensory cortex.

Fast P2X receptor-mediated excitatory postsynaptic current (EPSC) was identified in pyramidal neurones of layer II/III of somato-sensory cortex in acutely isolated slices obtained from the brain of 17- to 22-day-old rats. The EPSCs were elicited by electrical stimulation of vertical axons originating from layer IV-VI neurones at 0.1 Hz in the presence of bicuculline. When the glutamatergic EPSC was blocked by saturating concentrations of glutamate receptor inhibitors 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo-[f]-quinoxaline-7-sulphonamide (NBQX) and D-(-)-2-amino-5-phosphonopentanoic acid (D-AP5), a small EPSC component was recorded from 90 % of neurones tested. This residual EPSC was not affected by selective blockers of nicotinic (hexamethonium) or serotonin (N-(1-azabicyclo-[2.2.2]oct-3-yl)-6-chloro-4-methyl-3-oxo-3,4-dihydro-2H-1,4-benzoxazine-8-carboxamide hydrochloride, Y-25130) receptors, but it was reversibly inhibited by the antagonists of P2X receptors NF023 (8,8'-[carbonylbis(imino-3,1-phenylenecarbonylimino)]bis-1,3,5-naphthalene-trisulphonic acid), NF279 (8,8'-[carbonylbis (imino-4,1-phenylenecarbonylimino-4,1-phenylenecarbonylimino)]bis-1,3,5-naphthalene-trisulphonic acid) and PPADS (pyridoxal phosphate-6-azophenyl-2',4'-disulphonic acid). Application of ATP (10 microM) or alpha,beta-methylene ATP (10 microM) to pyramidal neurones, acutely isolated from cortical slices, evoked inward currents (30 to 200 pA) in 65 % of cells tested. The relative calcium/caesium permeability (P(Ca)/P(Cs)) of P2X receptors was 12.3 as estimated from the reversal potential of ATP-induced current measured at different extracellular calcium concentrations. We concluded that P2X purinoreceptors are activated during synaptic transmission in neocortex.

2-Amino-5-phosphonovalerate↗
PubMed2001

New channel blocker BIIA388CL blocks delayed rectifier, but not A-type potassium current in central neurons.

A new substance (R,S)-(3,4-dihydro-6,7-dimethoxyisoquinoline-1-yl)-2-cyclohexyl-N-(3,3-diphenylpropyl)-acetamide hydrochloride (BIIA388Cl), which demonstrates neuroprotective properties in animal models, was examined for its action on K(+) currents in acutely isolated rat hippocampal neurons using the patch-clamp/concentration clamp techniques in the whole-cell configuration. The delayed rectifier K(+)-current (I(DR)) was strongly inhibited by externally applied BIIA388Cl, while the transient A-current (I(A)) remained virtually unaffected. Block of I(DR) by the pre-applied BIIA388Cl was revealed as a rapid decay of the current indicating direct interaction of the drug with the open state of the channel. The removal of the block upon repolarization was also rapid (tau=22 ms). The dose-response relationship for the blocking action of BIIA388Cl revealed an IC(50) value of 300 nM for the peak I(DR), whereas the IC(50) value for I(DR) measured 300 ms after the onset of depolarization was 120 nM. The blocking action of BIIA388Cl on I(A) was at least 200 times less potent. These data allow us to conclude that BIIA388Cl is an effective and selective blocker of I(DR). This current is the main pathway for the loss of intracellular potassium by depolarized neurons. Selective obstruction of this pathway could be useful for neuroprotection.

Animals↗
PubMed2001

Modulation of ion channels in rat neurons by the constituents of Hypericum perforatum.

Despite almost forty years of widespread use of antidepressant drugs, their mode of action is still unknown. Hyperforin, a phloroglucinol derivative, is a major pharmacologically and therapeutically active constituent of Hypericum perforatum extract that is widely used as an herbal antidepressant drug. However, the mechanism or mechanisms of action of these naturally abundant, non-toxic extracts remain unclear. Enzymatically isolated patch-clamped rat central and peripheral neurons exposed to rapid changes in the composition of external medium (concentration clamp) were used in our experiments to investigate the modulation of the various voltage- and ligand-gated channels by hyperforin, as well as by other constituents of Hypericum perforatum. At nanomolar concentrations, hyperforin induced significant inhibition of various ion channels. In the case of P-type Ca2+ channels, we established that hyperforin acts via interaction with calmodulin or through calmodulin-activated pathways involving at least one second messenger. The results presented here indicate that multiple mechanisms and extract constituents may be involved in the antidepressant action of Hypericum extracts, and that they could also possess neuroprotective and analgesic effects.

Animals↗
PubMed2000

Hyperforin modulates gating of P-type Ca2+ current in cerebellar Purkinje neurons.

Whole-cell, patch-clamp recordings from acutely isolated cerebellar Purkinje neurons demonstrate a two-stage modulation of P-type high-voltage-activated (HVA) Ca2+ current by a constituent of St. John's wort, hyperforin (0.04-0.8 microM). The first stage of modulation was voltage dependent and reversible. It comprised slow-down of the activation kinetics and a shift in the voltage dependence of P-current to more negative voltages. Hyperforin (0.8 microM) shifted the maximum of the current/voltage (I/V) relationship by -8+/-2 mV. The second, voltage-independent stage of modulation was manifested as a slowly developing inhibition of P-current that could not be reversed within the period of study. Neither form of modulation was abolished by intracellular guanosine 5'-O-(2-thiodiphosphate) (GDPPS) or guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) or by strong depolarising pre-pulses, indicating that modulation via guanine nucleotide-binding proteins (G proteins) is not involved in the observed phenomenon. Calmidazolium (0.5 microM), an antagonist of the intracellular Ca2+-binding protein calmodulin significantly inhibited the hyperforin-induced shift of the IIV curve maximum and the slow-down of the activation kinetics. It did not, however, affect the delayed inhibition of P-current, indicating that the two stages of modulation are mediated by separate mechanisms.

Animals↗
PubMed1999

Hyperforin attenuates various ionic conductance mechanisms in the isolated hippocampal neurons of rat.

Effects of hyperforin, an acylphloroglucinol derivative isolated from antidepressive medicinal herb Hypericum perforatum (St. John's Wort), on voltage- and ligand-gated ionic conductances were investigated. Whole-cell patch clamp and concentration clamp techniques on acutely isolated hippocampal pyramidal neurons and on cerebellar Purkinje neurons of rat were used. At concentrations between 3 to 100 microM hyperforin induced a dose and time dependent inward current which completely stabilized within a few seconds. Although 1 microM hyperforin inhibited virtually all investigated conductances (GABA > or = I(Ca(N)) > I(Na) > I(Ca(P) > or = AMPA > or = I(K(A)) > NMDA > I(K(DR))), its effects on several of them could not be reversed by repeated washings. Dose response studies revealed that although AMPA induced current is inhibited by hyperforin in a competitive manner, these responses are not completely blocked by very high concentration of the agent. On the contrary, however, NMDA receptor-activated ionic conductance could be completely and uncompetitively inhibited by the agent. Taken together these observation not only reconfirm that hyperforin is a major neuroactive component of hypericum extracts but also demonstrate that this structurally unique and naturally abundant molecule is a potent modulation of mechanism involved in the control of neuronal ionic conductances. Various observed effects of hyperforin do not, however, seem to be mediated by one single molecular mechanism of action of the agent.

Animals↗
PubMed1999

The mechanism gated by external potassium and sodium controls the resting conductance in hippocampal and cortical neurons.

The excitation of densely packed mammalian central neurons is followed by a substantial transitory elevation of external K+ concentration. This phenomenon may have a different functional significance depending on how the resting membrane conductance mechanisms react to the changes in the gradient of these ions. We have found that in the hippocampal and cortex neurons of rat a large fraction of the membrane conductance in the vicinity of the resting potential is provided by the K+ permeability mechanism which is gated by external K+ and Na+. The responses of acutely isolated pyramidal neurons to rapidly altered external [K+] were investigated using the whole-cell patch clamp and concentration clamp techniques. Elevation of [K+]out induced a biphasic inward current at membrane potentials more negative than the reversal potential for K+ ions. This current consisted of an "instantaneously" increased leakage component and a slowly activated current (tau = 48 ms at 21 degrees C) designated below as I(deltaK). The latter demonstrated a first order activation kinetics with a remarkably high Q10 = 7.31. I(deltaK) was absent in the peripheral sensory neurons as well as in the Purkinje neurons. Slow activation of I(deltaK) was critically dependent on [Na+]out: substitution of the extracellular Na+ with choline chloride or Li+ led to the "instantaneous" reaction of the membrane to the changes in [K+]out. By slowing down potassium influx, I(deltaK) may be of importance in preserving densely packed pyramidal neurons from immediate excitation following rapid increases in [K+]out.

Animals↗
PubMed1998

A purinergic component of the excitatory postsynaptic current mediated by P2X receptors in the CA1 neurons of the rat hippocampus.

The pyramidal neurons in the CA1 area of hippocampal slices from 17- to 19-day-old rats have been investigated by means of patch clamp. Excitatory postsynaptic currents (EPSCs) were elicited by stimulating the Schaffer collateral at a frequency below 0.2 Hz. It was found that inhibition of glutamatergic transmission by 20 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 100 microM 2-amino-5-phosphonovaleric acid (D-APV) left a small component of the EPSC uninhibited. The amplitude of this residual EPSC (rEPSC) comprised 25 +/- 11% of the total EPSC when measured at a holding potential of -50 mV. The rEPSC was blocked by selective P2 blocker pyridoxal phosphate-6-azophenyl-2'-4'-disulphonic acid (PPADS) 10 microM and bath incubation with non-hydrolysable ATP analogues, ATP-gamma-S and alpha, beta-methylene-ATP at 50 and 20 microM, respectively. The rEPSC was dramatically potentiated by external Zn2+ (10 microM). In another series of experiments exogenous ATP was applied to the CA1 neurons in situ. An inward current evoked by ATP was inhibited by PPADS to the same extent as the rEPSC. It is concluded that, depending on membrane voltage, about one-fifth to one-quarter of the EPSC generated by the excitatory synaptic input to the hippocampal CA1 neurons of rat is due to the activity of P2X receptors.

2-Amino-5-phosphonovalerate↗
PubMed1996

NMDA receptor-mediated synapses between CA1 neurones: activation by ischaemia.

Using an in situ patch clamp in hippocampal CA1 mini-slices, we measured excitatory postsynaptic currents (EPSC) by varying the strength of the stimulus applied to the axons of CA3 neurones. The kinetics of the EPSC was initially independent of the stimulus strength. Post-ischaemic potentiation of the EPSC was observed 60-80 min after brief periods (10 min) of anoxia/aglycaemia. The decay of the EPSC slowed significantly in most of the examined neurones. In 11 of 17 cells the EPSC kinetics became dependent on stimulus strength: a slower decay corresponded to a stronger stimulus. This effect was not abolished by N-methyl-D-aspartate (NMDA) or a non-NMDA receptor blocker (D-2-amino-5-phosphonovaleric acid or 6-cyano-7-nitroquinoxaline-2,3-dione respectively) indicating the polysynaptic nature of the modified EPSC: transient ischaemia led to the long-term recruitment of previously inactive, possibly latent NMDA synapses between CA1 neurones.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1995

Latent N-methyl-D-aspartate receptors in the recurrent excitatory pathway between hippocampal CA1 pyramidal neurons: Ca(2+)-dependent activation by blocking A1 adenosine receptors.

When performed at increased external [Ca2+]/[Mg2+] ratio (2.5 mM/0.5 mM), temporary block of A1 adenosine receptors in hippocampus [by 8-cyclopentyltheophylline (CPT)] leads to a dramatic and irreversible change in the excitatory postsynaptic current (EPSC) evoked by Schaffer collateral/commissural (SCC) stimulation and recorded by in situ patch clamp in CA1 pyramidal neurons. The duration of the EPSC becomes stimulus dependent, increasing with increase in stimulus strength. The later occurring component of the EPSC is carried through N-methyl-D-aspartate (NMDA) receptor-operated channels but disappears under either the NMDA antagonist 2-amino-5-phosphonovaleric acid (APV) or the non-NMDA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). These findings indicate that the late component of the SCC-evoked EPSC is polysynaptic: predominantly non-NMDA receptor-mediated SCC inputs excite CA1 neurons that recurrently excite each other by predominantly NDMA receptor-mediated synapses. These recurrent connections are normally silent but become active after CPT treatment, leading to enhancement of the late component of the EPSC. The activity of these connections is maintained for at least 2 hr after CPT removal. When all functional NMDA receptors are blocked by dizocilpine maleate (MK-801), subsequent application of CPT leads to a partial reappearance of NMDA receptor-mediated EPSCs evoked by SCC stimulation, indicating that latent NMDA receptors are recruited. Altogether, these findings indicate the existence of a powerful system of NMDA receptor-mediated synaptic contacts in SCC input to hippocampal CA1 pyramidal neurons and probably also in reciprocal connections between these neurons, which in the usual preparation are kept latent by activity of A1 receptors.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1995

A1 adenosine receptors differentially regulate the N-methyl-D-aspartate and non-N-methyl-D-aspartate receptor-mediated components of hippocampal excitatory postsynaptic current in a Ca2+/Mg(2+)-dependent manner.

A1 adenosine receptors efficiently modulate the excitatory synaptic transmission in hippocampus. Here we report that in addition to previously known modulatory action on the synaptic efficacy, A1 adenosine receptors are also capable of regulating the relative contribution of N-methyl-D-aspartate receptor-mediated component of the excitatory postsynaptic current in CA3-CA1 excitatory synapses, in the rat. When applied externally, a selective A1 adenosine receptor antagonist, 8-cyclopentyl-1,3-dimethylxanthine, increases not only the amplitude of excitatory postsynaptic current but also the relative contribution of the N-methyl-D-aspartate receptor-mediated component of postsynaptic current recorded by in situ voltage clamp. This effect develops only at increased external Ca2+ concentration and also depends on the external Ca2+/Mg2+ ratio. The increased ratio of N-methyl-D-aspartate/non-N-methyl-D-aspartate components of excitatory postsynaptic current remains at a new level after the removal of 8-cyclopentyl-1,3-dimethylxanthine, even though the amplitude of excitatory postsynaptic current returns close to control value. These results indicate the existence of a mechanism that preferentially enhances the N-methyl-D-aspartate component of excitatory postsynaptic current when the A1 adenosine receptors are blocked and imprints the newly acquired ratio of corresponding excitatory postsynaptic current components.

2-Amino-5-phosphonovalerate↗
PubMed1994

Persistently enhanced ratio of NMDA and non-NMDA components of rat hippocampal EPSC after block of A1 adenosine receptors at increased [Ca2+]o/[Mg2+]o.

NMDA and non-NMDA receptor-mediated components of excitatory post-synaptic current (EPSC) were studied by in situ whole-cell voltage-clamp recordings in the CA1 field of rat hippocampus. We found that the amplitudes ratio of the NMDA to the non-NMDA components can be strongly increased by blocking A1 adenosine receptors. The necessary conditions for this effect are both, increased Ca2+ and lowered Mg2+ in the external medium. The so achieved increase in the NMDA/non-NMDA ratio of EPSC components is irreversible and no longer depends on the activity of A1 adenosine receptors.

Animals↗
PubMed1994

Possible functional role of diadenosine polyphosphates: negative feedback for excitation in hippocampus.

Diadenosine polyphosphates (Ap4A and Ap5A) are present in secretory granules of chromaffin cells as well as in the rat brain synaptic terminals. Their contribution to the exocytosis of the total synaptosomal content is considerable, ranging from 7% to 12%. Ap4A and Ap5A are released from synaptosomes in a Ca(2+)-dependent manner. There are indications on the high affinity of diadenosine polyphosphates to P2 receptors, but their action on P1 receptors remains unclear. Here we report that both substances induce a blocking action on excitatory synaptic transmission in the rat hippocampus. This action is elicited via the A1 (subclass of P1) receptors and differs in some respects from the action of adenosine.

Animals↗
PubMed1993

Glutamate and theta-rhythm stimulation selectively enhance NMDA component of EPSC in CA1 neurons of young rats.

Using in situ whole-cell patch clamp of hippocampal CA1 pyramidal neurons we demonstrate that glutamate initiates processes resulting in an increase in the amplitude of the excitatory post-synaptic current (EPSC). In adult animals both, NMDA and non-NMDA components of the EPSC increase in parallel. In young animals only the NMDA component is increased. A similar enhancement of the EPSC can be achieved by the stimulation of excitatory synaptic inputs to CA1 neurons with the frequency of the theta-rhythm. EPSCs remain enhanced for more than 60 min. The selective enhancement of the NMDA component in young animals is inhibited by preincubation of slices with the NO-synthase blocker, N omega-nitro-L-arginine (NA) or by the NO-scavenger, hemoglobin.

Aging↗