Pyridoxine 5'-phosphate oxidase, not pyridoxal kinase, involves in long-term potentiation induction in the rat dentate gyrus.

In this study, we investigated whether the expression of pyridoxal kinase (PLK) and pyridoxine-5'-phosphate oxidase (PNPO) are altered following long-term potentiation (LTP) induction, and whether Tat-PLK and Tat-PNPO transductions affect LTP induction and paired-pulse responses in the rat dentate gyrus (DG). PNPO immunoreactivity was markedly increased in dentate granule cells after the induction of LTP, but that of PLK was not. Tat-PNPO (20 and 200 microg/kg), but not Tat-PLK or vitamin B6 precursors, treatments, increased the efficiency of high frequency stimulus-induced potentiation of populations spike amplitude when compared with saline-, or Tat-protein-treated animals. These changes correlated with the alterations in PNPO activity and its immunoreactivity. In addition, Tat-PNPO transduction increased paired-pulse facilitation but had no effect on the fast and late paired-pulse inhibitions. These findings suggest that PNPO may play a role in activity-dependent regulation of PLP level in the brain, which is involved in LTP induction and paired-pulse facilitation but not in enhancement of GABAergic inhibition.

Pyridoxal-5'-phosphate phosphatase/chronophin inhibits long-term potentiation induction in the rat dentate gyrus.

Pyridoxal-5'-phosphate (PLP)-phosphatase/chronophin (PLPP/CIN) directly dephosphorylates actin-depolymerizing factor (ADF)/cofilin as well as PLP. Although PLPP/CIN plays a role in the regulation of F-actin and vitamin B(6) metabolism, there is no direct evidence to support a correlation between PLPP/CIN and F-actin polymerization during long-term potentiation (LTP) induction. In this study, we investigated whether the expression of PLPP/CIN is altered following LTP induction, and whether Tat-PLPP/CIN transduction affects LTP induction in the rat dentate gyrus (DG). PLPP/CIN immunoreactivity was markedly decreased in dentate granule cells after the induction of LTP. Tat-PLPP/CIN transduction (20 and 200 microg/kg) decreased the efficiency of high frequency stimulus-induced potentiation of populations spike amplitude as compared to saline or Tat-protein-treated animals. The PLPP/CIN protein level showed an inverse correlation with phosphorylated ADF/cofilin levels and F-actin content. These findings suggest that PLPP/CIN-mediated actin dynamics may play an important role in the changes of morphological properties (dendritic spine reorganization) of the hippocampus in LTP.

Upregulated TWIK-related acid-sensitive K+ channel-2 in neurons and perivascular astrocytes in the hippocampus of experimental temporal lobe epilepsy.

PURPOSE: To identify the modulation of Tandem of P-domains in a weak inwardly rectifying K(+) channel (TWIK)-related acid-sensitive K(+) (TASK)-2 channel expressions in epilepsy, we conducted a comparative analysis of TASK-2 channel immunoreactivity in the hippocampus of a pilocarpine-induced rat epilepsy model. METHODS: We performed and immunohistochemical study for TASK-2 and double immunofluorescent staining for TASK-2 and glial fibrillary acidic protein (GFAP) in the rat hippocampus of pilocarpine-induced epilepsy models. RESULTS: In control animals, TASK-2 immunoreactivity was strongly detected in CA1-3 pyramidal layers and dentate granule cell layer. After status epilepticus (SE), TASK-2 immunoreactivity was increased in dentate granule cell layer and CA3 pyramidal cell layer, whereas its immunoreactivity was reduced in CA1 pyramidal cell layer. In addition, TASK-2 immunoreactivity is gradually increased in perivascular regions following SE. Double immunofluorescent study revealed that the enhancement of TASK-2 immunoreactivity in perivascular regions is caused by increase in the number of TASK-2 immunoreactive endfeet of perivascular astrocytes. DISCUSSION: Our findings suggest that elevated TASK-2 immunoreactivity in neurons may contribute to rapid adaptive responses (presumably for extracellular alkalinization), which result in hyperpolarization and regulate seizure activity. In contrast, upregulated TASK-2 immunoreactivity in perivascular regions may be involved in abnormalities of blood flow regulation or brain-blood barrier impairment. These changes may contribute to acquisition of the properties of the epileptic hippocampus.

Age-dependent pathogenesis of murine gammaherpesvirus 68 infection of the central nervous system.

Gammaherpesvirus infection of the central nervous system (CNS) has been linked to various neurological diseases, including meningitis, encephalitis, and multiple sclerosis. However, little is known about the interactions between the virus and the CNS in vitro or in vivo. Murine gammaherpesvirus 68 (MHV-68 or (gamma)HV-68) is genetically related and biologically similar to human gammaherpesviruses, thereby providing a tractable animal model system in which to study both viral pathogenesis and replication. In the present study, we show the successful infection of cultured neuronal cells, microglia, and astrocytes with MHV-68 to various extents. Upon intracerebroventricular injection of a recombinant virus (MHV-68/LacZ) into 4-5-week-old and 9-10-week-old mice, the 4-5-week-old mice displayed high mortality within 5-7 days, while the majority of the 9-10-week-old mice survived until the end of the experimental period. Until a peak at 3-4 days post-infection, viral DNA replication and gene expression were similar in the brains of both mouse groups, but only the 9-10-week-old mice were able to subdue viral DNA replication and gene expression after 5 days post-infection. Pro-inflammatory cytokine mRNAs of tumor necrosis factor-alpha, interleukin 1beta, and interleukin 6 were highly induced in the brains of the 4-5-week-old mice, suggesting their possible contributions as neurotoxic factors in the agedependent control of MHV-68 replication of the CNS.

Anticonvulsant characteristics of pyridoxyl-gamma-aminobutyrate, PL-GABA.

GABA is the major inhibitory neurotransmitter in the central nervous system, and its concentration in the brain in associated with a variety of neurological disorders, including seizures, convulsions, and epilepsy. The concentration of GABA is modulated by the pyridoxal-5'-phosphate (PLP)-dependent enzymes, GAD and GABA-T. In this study, we generated pyridoxyl-gamma-aminobutyrate (PL-GABA), a novel GABA analogue composed of pyridoxyl and GABA, and have also characterized its anticonvulsant and pharmacological functions in vitro. The results of biodistribution studies revealed that PL-GABA is capable of crossing the blood-brain barrier. PL-GABA evidenced anticonvulsant activity in a wide range of epilepsy models, some of which were electrically-based (MES seizures) and some chemically-based (bicuculline, pentylenetetrazol (PTZ), picrotoxine, 3-mercaptopropionic acid). Following a timed subcutaneous administration of PTZ to mice, PL-GABA consistently increased the latencies to first twitch and clonus. In addition, PL-GABA displayed no signs of tolerance after subchronic (10 day) treatment. PL-GABA appears to exert its anticonvulsant effects by influencing seizure spread and by raising the seizure threshold. Therefore, our results indicate that PL-GABA exerts a broad-spectrum anticonvulsant effect, and identify the potential for reduced PL-GABA tolerance as an additional positive profile for novel antiepileptic drugs.

The expression of somatostatin receptors in the hippocampus of pilocarpine-induced rat epilepsy model.

During the course of this study, we sought examine whether the expression of somatostatin receptors (SSTRs) is altered in the hippocampus following pilocarpine-induced status epilepticus (SE) in order to understand the role/function of SSTRs in the hippocampus after epileptogenic insults. SSTR1 and SSTR4 immunoreactivities were increased in the hippocampus at 1 week after SE. At 4 weeks after SE, SRIF1-family (SSTR 2A, SSTR2B, and SSTR5) immunoreactivity was increased only in neuropil. Both SSTR2A and 2B immunoreactivities were increased in CA2-3 pyramidal cells. However, SSTR3 and SSTR4 immunoreactivities were reduced in the CA1 pyramidal cells of epileptic rat due to neuronal loss. In addition, SSTR5 immunoreactivity was reduced in CA2 pyramidal cells and various interneurons. Both SSTR2B and SSTR4 immunoreactivities were increased within microglia following SE. Our findings suggest that increases in neuron-glial SSTR expressions may be closely related to the enhanced inhibition of the dentate gyrus and regulation of reactive microgliosis in the hippocampus of a pilocarpine model of temporal lobe epilepsy.

Hyperthermic seizure induces persistent alteration in excitability of the dentate gyrus in immature rats.

Febrile seizure (FS) is the most common type of seizure that occurs during early childhood. It has been proposed that atypical FS (prolonged, multiple, or lateralized) results in the development of recurrent complex partial seizures accompanied by Ammon's horn sclerosis or mesial temporal sclerosis, which is the most common of the intractable epilepsy. To elucidate the characteristics of epileptogenesis or acquired epilepsy following FS, we performed prospective long-term studies using hyperthermia-induced seizure model. Rat pups (postnatal 11 day old) were induced to hyperthermia (41-43 degrees C in core temperature) by exposure to a 175 W mercury vapor lamp. Six-nine weeks after hyperthermic seizure, the dentate gyrus showed impairments of paired-pulse inhibitions and excitability ratio. In addition, newly generated granule cells and synaptogenesis were observed in this region. Ten-twelve weeks after hyperthermic seizure, animals (approximately 68%) showed electroencephalographic seizure activity with increased VGLUT-1 immunoreactivity in the dentate gyrus. Parvalbumin immunoreactivity was markedly reduced in the hilus. These findings indicate that in this model the epileptogenic changes in the dentate gyrus may be based on the persistent alterations in excitability via neurogenesis, synaptogenesis, and impaired GABA(B) receptor-mediated inhibition.

Voltage-gated Na+ channel II immunoreactivity is selectively up-regulated in hippocampal interneurons of seizure sensitive gerbils.

In the present study, we investigated the distribution of voltage-gated Na(+) channels (VGSCs) in the normal and epileptic hippocampus of gerbils (a genetic epilepsy model) in order to confirm the relationship between VGSC and seizure activity in these animals. There was no difference of VGSC I immunoreactivity in the hippocampus between seizure-resistant (SR) and seizure sensitive (SS) gerbils. VGSC II immunoreactivity was rarely detected in the perikarya of principal neurons and interneurons in the SR gerbil hippocampus. However, in the SS gerbil hippocampus, VGSC II immunoreactivity was densely observed in the somata of interneurons located in the stratum radiatum and stratum lacunosum-moleculare. Double immunofluorescent study showed immunoreactivity for calretinin (approximately 80% in VGSC II-positive neurons) or calbindin D-28k (approximately 20% in VGSC II-positive neurons) in VGSC II-immunoreactive neurons. VGSC II-immunoreactive neurons did not show parvalbumin immunoreactivity. These findings suggest that seizure activity in SS gerbils may be related to the selective hyperactivation of interneurons in stratum lacunosum-moleculare via the up-regulation of VGSC II expression, which leads to the disinhibition of CA1 pyramidal cells.

Up-regulation of P/Q-type voltage-gated Ca2+ channel immunoreactivity within parvalbumin positive neurons in the rat hippocampus following status epilepticus.

To identify the roles of VGCC subtypes in damages/impairs of inhibitory transmission during epileptogenesis, we investigated temporal- and spatial-specific alterations in voltage-gated Ca(2+) channel (VGCC) immunoreactivities within parvalbumin (PV, a Ca(2+) binding protein) positive neurons in the rat hippocampus following status epilepticus (SE). Compared to controls, only P/Q-type (alpha1A) VGCC immunoreactivity was enhanced in PV positive neurons at the early point following SE. The alteration in P/Q-type (alpha1A) VGCC immunoreactivity showed an inverse proportionality to that in PV immunoreactivity in the dentate gyrus and the CA1 region. These findings suggest that SE may induce prolonged up-regulation in P/Q-type VGCC expression within PV positive neurons.

Seizure activity affects neuroglial Kv1 channel immunoreactivities in the gerbil hippocampus.

In order to confirm the species-specific distribution of voltage-gated K(+) (Kv) channels and the definitive relationship between their immunoreactivities and seizure activity, we investigated Kv1 channel immunoreactivities in the hippocampus of seizure resistant (SR) and seizure sensitive (SS) gerbils. There was distinct difference of the Kv1 channel subtypes immunoreactivity in the hippocampi in both SR and SS gerbils. Kv1.1, Kv1.2, Kv1.3, Kv1.4, and Kv1.6 immunoreactivities in the SS gerbil hippocampus were lower than that in the SR gerbil hippocampus. However, Kv1 immunoreactivities were obviously presented in astrocyte within the stratum radiatum of the CA1 region of pre-seizure SS gerbil hippocampus. Following seizure-onset, Kv1 immunoreactivities (except Kv1.5) were markedly elevated, whereas their immunoreactivites in astrocytes were down-regulated. Therefore, the present study demonstrates that seizure activity may distinctly affect neuroglial Kv1 immunoreactivities in the gerbil hippocampus.

Seizure activity selectively reduces 5-HT1A receptor immunoreactivity in CA1 interneurons in the hippocampus of seizure-prone gerbils.

Since the correlation between the serotonin (5-hydroxytryptamine, 5-HT) system and seizure activity remains to be clarified, we investigated the 5-HT system in the hippocampus of seizure-resistant (SR) and seizure-sensitive (SS) gerbils. There was no difference of the 5-HT system in the hippocampi of young animals (predisposed and juvenile gerbils) in both SR and SS gerbils. 5-HT immunoreactivity in the dorsal raphe nucleus and the median raphe nucleus was also similarly detected in both animal groups. As compared to SR adult gerbils, only 5-HT1A receptor immunoreactivity was selectively reduced in CA1 interneurons within SS adult gerbils. (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin (a 5-HT1A receptor agonist, 1 and 2 mg/kg) markedly reduced paired-pulse inhibition in the CA1 region of SS adult gerbils only. These findings suggest that the selective reduction in 5-HT1A receptor expression on CA1 interneurons of SS adult gerbil may not be developmental defects, but be an acquired compensatory change induced by repeated seizure activity.

Up-regulated astroglial TWIK-related acid-sensitive K+ channel-1 (TASK-1) in the hippocampus of seizure-sensitive gerbils: a target of anti-epileptic drugs.

In order to identify the modulation of TASK (TWIK-related Acid-Sensitive K(+)) channel expressions in epilepsy, we conducted a comparative analysis of TASK channel immunoreactivities in the hippocampus of seizure-resistant (SR) and seizure-sensitive (SS) gerbils. There was no difference of the TASK-1 and TASK-2 channel expressions in the hippocampi of young SR and SS gerbils (1-2 months old). In adult SS gerbil hippocampus, TASK-1 immunoreactivity in astrocytes was higher than that in adult SR gerbil hippocampus. After seizures, TASK-1 immunoreactivity was significantly down-regulated in astrocytes of the SS gerbil hippocampus. In addition, various anti-epileptic drugs selectively affect TASK-1 immunoreactivity in astrocytes of the SS gerbil hippocampus. Gabapentin, lamotrigine, topiramate and valproic acid reduced the number of TASK-1(+) astrocytes in the hippocampus to 10-25% of that in saline-treated SS adult gerbils, whereas carbamazepine and vigabatrin decreased to approximately 50%. Therefore, the present study demonstrates that up-regulated TASK-1 immunoreactivity in astrocytes may be involved in the seizure activity of SS adult gerbils and suggests that the astroglial TASK-1 channel may be a target for epilepsy therapeutics.

The co-treatments of vigabatrin and P2X receptor antagonists protect ischemic neuronal cell death in the gerbil hippocampus.

During transient global ischemia, the excessive accumulation of intracellular Ca2+ induced by several episodes triggers delayed neuronal death within the vulnerable CA1 region of the hippocampus after ischemia-reperfusion insults. Although P2X receptors provide an additional source of Ca2+ entry, little data are available that these receptors could modulate the performance of the ischemic neuronal death. Therefore, we investigated the roles of the P2X receptor in the ischemic neuronal damage associated with various sequelae of transient ischemia, and the effects of their antagonist on the ischemic insults. As the results, ischemic insults increased P2X receptor expression in the gerbil hippocampus. Neither vigabatrin (VGB) nor P2X receptor antagonists (suramin, pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid) protected against the delayed neuronal death in the CA1 region of the hippocampus after ischemia. However, the co-treatments of VGB and P2X receptor antagonists effectively prevent ischemia-induced neurodegeneration. Therefore, these findings suggest that blockade of the P2X receptor accompanied by activation of GABAergic inhibition may play an important role in the neuroprotection against ischemic insults.

The selective effects of somatostatin- and GABA-mediated transmissions on voltage gated Ca2+ channel immunoreactivity in the gerbil hippocampus.

To identify whether altered expressions of voltage gated Ca(2+) channel (VGCC) are linked to inhibitory transmission abnormalities in the gerbil hippocampus, we investigated the effects of GABA receptor or somatostatin receptor (SST) antagonists/agonists on VGCC immunoreactivity in vivo. VGCC immunoreactivities in the hippocampus were significantly higher in seizure sensitive (SS) gerbils than in seizure resistant (SR) gerbils. P/Q-type VGCC immunoreactivity in the gerbil hippocampus was reduced by enhancement in GABA(A) and GABA(B) receptor-mediated transmission, but not by SST-mediated transmission. N-type VGCC immunoreactivity was reduced only by a SST agonist, whereas L-type (alpha1C) VGCC immunoreactivity was reduced only by a GABA(A) receptor agonist, and L-type (alpha1D) VGCC immunoreactivity was modulated by the GABA(B) receptor acting drugs. These findings provide a comprehensive description of the differential responses of VGCC subunits to alteration in GABAergic or somatostatinergic transmission. These findings also suggest that up-regulated VGCC immunoreactivity may be consequence of the neuronal excitability caused by a reduction in inhibitory neurotransmission in the gerbil hippocampus.

Differential paired-pulse responses between the CA1 region and the dentate gyrus are related to altered CLC-2 immunoreactivity in the pilocarpine-induced rat epilepsy model.

The epileptic hippocampus shows differential paired-pulse responses between the dentate gyrus and the CA1 region. However, little data are available to explain this phenomenon. In the present study, we identified the relationship between regional differences of paired-pulse response and voltage gated Cl(-) channel 2 (CLC-2)/vesicular GABA transport (VGAT) expression in a pilocarpine-induced rat model. During epileptogenic periods, paired-pulse inhibitions in the dentate gyrus and the CA1 region were markedly reduced. After recurrent seizure onset, paired-pulse inhibition in the dentate gyrus was markedly enhanced, while that in the CA1 region more reduced. Unlike VGAT, CLC-2 immunoreactivity was markedly reduced in the hippocampus during epileptogenic periods and was re-enhanced only in the dentate gyrus after recurrent seizure onset. Linear regression analysis showed an inverse proportional relationship between alterations in CLC-2 immunoreactivity and changes in normalized population spike amplitude ratio within the CA1 region and the dentate gyrus. Therefore, our findings suggest that the regionally specific alterations in CLC-2 immunoreactivity after SE may determine the properties of paired-pulse responses in the hippocampus of the pilocarpine-induced rat epilepsy model.

Altered expression of K+ -Cl- cotransporters affects fast paired-pulse inhibition during GABA receptor activation in the gerbil hippocampus.

K+ -Cl- cotransporter (KCC) plays an important role in maintaining neuronal activity. However, the effect of seizure activity or pharmacological manipulation of GABAergic transmission on KCC expression remains to be clarified. Therefore, the present study was performed to investigate whether seizure activity or GABA receptor agonist treatment changes KCC expression in the gerbil hippocampus. Furthermore, the effect of blockade of KCC on inhibitory transmission in the dentate gyrus was identified following applications of GABA receptor agonists. The distribution of KCC immunoreactivity in the hippocampus was similarly detected between seizure-resistant (SR) and seizure-sensitive (SS) gerbils. Baclofen (a GABAB receptor agonist) treatment markedly increased KCC expression in the gerbil hippocampus. Baclofen treatment significantly reduced paired-pulse inhibition in the dentate gyrus. Furosemide (a KCC inhibitor) treatment amplified the effect of baclofen on paired-pulse responses. In contrast, muscimol (a GABAA receptor agonist) treatment reduced KCC expression. Enhanced paired-pulse inhibition by muscimol treatment was not affected by furosemide treatment. These findings suggest that seizure activity in the gerbil may not affect KCC expression in the hippocampus. In addition, altered KCC immunoreactivity induced by baclofen or muscimol may play an important role in maintaining or regulating inhibitory transmission during GABA receptor activation.

Effects of GABAergic transmissions on the immunoreactivities of calcium binding proteins in the gerbil hippocampus.

Although reduced calcium binding protein (CBP) immunoreactivities in the epileptic hippocampus have been well established, it has been controversial that these changes may directly indicate neuronal degeneration. In the present study, therefore, we investigated CBP expressions in the gerbil hippocampus following treatment with gamma-aminobutyric acid (GABA) receptor antagonists in order to assess whether altered CBP expressions are the result of either abnormal excitation or indicative of neuronal damage/degeneration. Seizure-sensitive (SS) gerbils showed a loss/decline of CBP immunoreactivities in some hippocampal neurons as compared with seizure-resistant (SR) gerbils. In muscimol (GABA(A) receptor agonist) treated SS gerbils, expression levels of CBP were enhanced as compared with saline-treated SS gerbils. Bicuculline (a GABA(A) receptor antagonist) treatment markedly reduced CBP immunoreactivities in hippocampal neurons of the SR gerbil. Baclofen (a GABA(B) receptor agonist) treatment increased CBP immunoreactivities in the hippocampus of SS gerbils, although its effect was lower than that of muscimol treatment. Moreover, phaclofen (GABA(B) receptor antagonist) treated SR gerbil showed reduction in calbindin D-28K immunoreactivity, not parvalbumin immunoreactivity, in the hippocampus. These findings therefore suggest that reduced CBP immunoreactivities may be the consequence of abnormal discharge caused by loss of GABAergic inhibition rather than an indication of the neuronal damage/degeneration.

The effect of topiramate on GABA(B) receptor, vesicular GABA transporter and paired-pulse inhibition in the gerbil hippocampus.

To extend our understanding of the properties of topiramate (TPM), we investigated the effect of TPM on GABAergic transmission in the dentate gyrus of gerbil. TPM treatment (> or = 40 mg/kg) dramatically decreased GABA(B)R2, not GABA(B)R1, immunoreactivity in hilar interneurons. In contrast, TPM treatment increased vesicular GABA transporter immunoreactivity and the paired-pulse inhibition in the dentate gyrus of seizure prone gerbils. Furthermore, TPM effectively prevented the reduction of paired-pulse inhibition induced by baclofen treatment. These findings suggest that TPM may enhance GABA release in the dentate gyrus of gerbils by down-regulation of GABA(B) autoreceptor expression. Therefore, these properties of TPM may be another possible antiepileptic effect, which plays an important role in preventing the spread of seizure activity without proconvulsive effects.

Differential effects of vigabatrin and zonisamide on the neuropeptide Y system in the hippocampus of seizure prone gerbil.

Changed neuropeptide Y (NPY) system in the hippocampus has been reported in various experimental epileptic models. However, there have been little data concerning the alteration in the NPY system in the epileptic hippocampus following treatment of anti-epileptic drugs (AEDs). In the present study, therefore, we performed analyses of effects of vigabatrin (VGB) and zonisamide (ZNS) treatment on the NPY system in the hippocampus of the seizure sensitive (SS) gerbils. In SS gerbil, NPY immunoreactivity in the hippocampus was lower than that in seizure resistant gerbil. Following VGB treatment, the number of NPY immunoreactive neurons and NPY mRNA expression were increased in the hilus and the hippocampus proper. In contrast, ZNS treatment markedly elevated only the density of NPY immunoreactive fibers in the dentate gyrus, not in the hippocampus proper, as compared with saline-treated animals. These patterns were observed in the dose-dependent manners. These findings suggest that AEDs treatments may distinctly affect the NPY system in the SS gerbil hippocampus.

The effect of P2X receptor activity on GABAA receptor-mediated inhibition in the gerbil hippocampus.

In the present study, to elucidate the effect of altered P(2)X receptor transmission on GABA(A) receptor expression and its transmission, we studied the morphological and electrophysiological responses of GABA(A) receptor in the gerbil hippocampus following P(2)X receptor antagonist/agonist treatment. Suramin or pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) treatment did not affect GABA(A) receptor immunoreactivities and paired-pulse responses in the gerbil hippocampus. In addition, ATP treatment did not significantly affect population spike amplitude ratios and EPSP slope ratios in the gerbil dentate gyrus. Co-application, but not pretreatment, of PPADS or suramin enhanced the effect of muscimol on paired-pulse inhibition in the dentate gyrus. In contrast, co-application of ATP reduced the effect of muscimol in the dentate gyrus. These findings indicate that the blockade of P(2)X receptor did not affect GABA(A) receptor immunoreactivities, and P(2)X receptor may modulate GABA(A) receptor-mediated inhibition when in co-activation with GABA(A) receptor. Therefore, our findings suggest that the relationship between GABA(A) receptor and P(2)X receptor may not be reciprocal, although GABA(A) receptor activity affects P(2)X receptor functionality and its expression.