Biomedical subjects

N Akaike

Publications and source records attributed to N Akaike.

At least 55 records · Page 3Linked to original sources

PubMed1999

Regulation of intracellular chloride by cotransporters in developing lateral superior olive neurons.

The regulatory mechanisms of intracellular Cl- concentration ([Cl-]i) were investigated in the lateral superior olive (LSO) neurons of various developmental stages by taking advantage of gramicidin perforated patch recording mode, which enables neuronal [Cl-]i measurement. Responses to glycine changed from depolarization to hyperpolarization during the second week after birth, resulting from [Cl-]i decrease. Furosemide equally altered the [Cl-]i of both immature and mature LSO neurons, indicating substantial contributions of furosemide-sensitive intracellular Cl- regulators; i.e., K+-Cl- cotransporter (KCC) and Na+-K+-Cl- cotransporter (NKCC), throughout this early development. Increase of extracellular K+ concentration and replacement of intracellular K+ with Cs+ resulted in [Cl-]i elevation at postnatal days 13-15 (P13-P15), but not at P0-P2, indicating that the mechanism of neuronal Cl- extrusion is sensitive to both furosemide and K+-gradient and poorly developed in immature LSO neurons. In addition, removal of extracellular Na+ decreased [Cl-]i at P0-P2, suggesting the existence of extracellular Na+-dependent and furosemide-sensitive Cl- accumulation in immature LSO neurons. These data show clearly that developmental changes of Cl- cotransporters alter [Cl-]i and are responsible for the switch from the neonatal Cl- efflux to the mature Cl- influx in LSO neurons. Such maturational changes in Cl- cotransporters might have the important functional roles for glycinergic and GABAergic synaptic transmission and the broader implications for LSO and auditory development.

Animals↗

PubMed1999

Functional reconstitution of a heteromeric cyclic nucleotide-gated channel of Caenorhabditis elegans in cultured cells.

The tax-4 and tax-2 genes of Caenorhabditis elegans are essential for normal olfaction, gustation, and thermosensation, suggesting that they have a role in sensory transduction. The predicted products of these genes are similar to the cyclic nucleotide-gated (CNG) channel subunits used in vertebrate vision and olfaction: TAX-4 is highly related to those alpha subunits, while TAX-2 is most closely related to the beta subunits of the rod phototransduction channels. TAX-4 has previously been shown to form a highly sensitive cGMP-gated channel when expressed in human HEK293 cells. Here we show that TAX-4 and TAX-2 form a heteromeric channel when expressed in HEK293 cells, but TAX-2 does not form a channel on its own. Since these genes are expressed in the same neurons, most of the native channels in C. elegans are likely to be hetero-oligomers of TAX-4 and TAX-2 subunits, with TAX-4 as the alpha subunit and TAX-2 acting as a modifying beta subunit. The heteromeric TAX-4/TAX-2 channel is 25-fold less sensitive to cGMP than the TAX-4 channel, but it remains highly selective for cGMP over cAMP. The heteromeric channel and the TAX-4 homomeric channel differ in their blockage by divalent cations and in their single channel properties. These results suggest that cGMP is used as the second messenger during sensory signal transduction in C. elegans, and that distinct roles for alpha and beta subunits of CNG channels are conserved in vertebrate and invertebrate signal transduction.

Animals↗

PubMed1999

Modulation of the glycine response by Ca2+-permeable AMPA receptors in rat spinal neurones.

1. In acutely isolated rat sacral dorsal commisural nucleus (SDCN) neurones, application of kainate (KA) reversibly potentiated glycine-evoked Cl- currents (IGly) in a concentration-dependent manner. 2. The cellular events underlying the interaction between non-NMDA receptors and glycine receptors were studied by using nystatin-perforated patch and cell-attached single-channel recording modes. 3. The action of KA was not accompanied by a shift in the reversal potential for IGly. In dose-response curves, KA potentiated IGly without significantly changing glycine binding affinity. 4. GYKI 52466 blocked while NS-102 had no effect on the KA-induced potentiation of IGly. 5. The potentiation was reduced when KA was applied in a Ca2+-free extracellular solution or in the presence of BAPTA AM, and was independent of the activation of voltage-dependent Ca2+ channels. 6. Pretreatment with KN-62, a selective Ca2+-calmodulin-dependent protein kinase II (CaMKII) inhibitor, abolished the action of KA. Inhibition of calcineurin converted the KA-induced potentiation to a sustained one. 7. Single-channel recordings revealed that KA decreased the mean closing time of glycine-gated single-channel activity, resulting in an increase in the probability of channel opening. 8. It is proposed that Ca2+ entry through AMPA receptors modulates the glycine receptor function via coactivation of CaMKII and calcineurin in SDCN neurones. This interaction may provide a new postsynaptic mechanism for control of inhibitory synaptic signalling and represent one of the important regulatory mechanisms of spinal nociception.

Animals↗

PubMed1999

Functional properties of ionotropic glutamate receptor channels in rat sacral dorsal commissural neurons.

Nystatin perforated patch and conventional whole-cell recording configurations were used to characterize the properties of ionotropic glutamate receptor (GluR) channels in neurons freshly dissociated from the rat sacral dorsal commissural nucleus (SDCN). L-Glutamate (Glu), N-methyl-D-aspartate (NMDA), quisqualate (QA), alpha-amino-3-hydroxy-5-methyl-4-isoxazoleprop ionate (AMPA) and kainate (KA) applied via a Y-tube produced inward currents at -44 mV which increased in a concentration-dependent manner; they desensitized when induced at higher concentrations except for the KA-induced current (IKA). (1S-3R)1-amino-cyclopentane-1,3-dicarboxylate (1S-3R-ACPD) evoked no response. The EC50 and Hill coefficient (nH) values of the GluR responses were 3.3 x 10(-5) M, 0.74 for Glu; 9.0 x 10(-5) M, 0.83 for NMDA; 6.4 x 10(-7) M, 1.30 for QA; 1.3 x 10(-4) M, 1.10 for AMPA and 9.6 x 10(-5) M, 1.30 for KA, respectively. The reversal potentials of the GluR responses were all near 0 mV. The 6-Cyano-7-nitroquinoxaline-2-3-dione (CNQX) and D-2-amino-5-phosphonovalerate (D-APV) suppressed the non-NMDA and NMDA responses in a concentration-dependent manner, respectively. Cyclothiazide strongly potentiated both KA- and AMPA-induced responses while concanavalin A potentiated both the responses to a much lesser degree. NS-102 produced no significant effect on either KA- or AMPA-activated currents, while GYKI 52466 reversibly blocked both the currents. The Ca2+ permeabilities (PCa/PCs) of the NMDA and AMPA receptor channels were 8.33 and 1.23, respectively. In addition, the current-voltage (I-V) relationship of IKA showed little rectification. There was a poor correlation between the Ca2+ permeability and the shape of the I-V curves of IKA. These results suggest that rat SDCN neurons possess NMDA and non-NMDA receptor channels, and express AMPA type receptors with unique properties (slow desensitization to AMPA, high Ca2+ permeability but lack of inward rectification). These ionotropic receptor channels may play important roles in mediating and regulating pelvic visceral information including nociception.

Animals↗

PubMed1999

Alpha2-adrenoceptor-mediated enhancement of glycine response in rat sacral dorsal commissural neurons.

The effect of noradrenaline on the glycine response was investigated in neurons acutely dissociated from the rat sacral dorsal commissural nucleus using nystatin perforated patch recording configuration under voltage-clamp conditions. Noradrenaline reversibly potentiated the 10(-5)M glycine-induced Cl- current in a concentration-dependent manner. Single channel recordings in a cell-attached mode revealed that noradrenaline decreased the closing time of the glycine-activated channel activity. Noradrenaline neither changed the reversal potential of the glycine response nor affected the affinity of glycine to its receptor. Clonidine mimicked and yohimbine blocked the noradrenaline action on glycine response. N-[2(methylamino)ethyl]-5-isoquinoline sulfonamide dihydrochloride, protein kinase A inhibitor, mimicked the effect of noradrenaline on glycine response. Noradrenaline failed to affect the glycine response in the presence of these intracellular cyclic AMP and protein kinase A modulators. However, noradrenaline further enhanced the glycine response even in the presence of phorbol-12-myristate-13-acetate and chelerythrine, a protein kinase C inhibitor. Pertussis toxin treatment for 6-8 h blocked the noradrenaline facilitatory effect on the glycine response. In addition, noradrenaline potentiated the strychnine-sensitive postsynaptic currents evoked in a slice preparation of sacral dorsal commissural nucleus. These results suggest that the activation of alpha2-adrenoceptor by noradrenaline coupled with pertussis toxin-sensitive G-proteins reduces intracellular cyclic AMP formation through the inhibition of adenyl cyclase. The reduction of cyclic AMP decreases the protein kinase A activity, thus resulting in the potentiation of the glycinergic inputs to the sacral dorsal commissural neurons. It is thus feasible that the noradrenergic input to the sacral dorsal commissural nucleus modulates such nociceptive signals as pain by intracellular enhancing the glycine response.

1-Methyl-3-isobutylxanthine↗

PubMed1999

Excitatory amino acid responses in relay neurons of the rat lateral geniculate nucleus.

Responses to glutamate receptor agonists were recorded from identified relay neurons in the dorsal lateral geniculate nucleus of the rat, using the nystatin-perforated patch-clamp technique. Rapid application of glutamate, N-methyl-D-aspartate, (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and kainate induced inward currents at a holding potential of -44 mV. The responses to low concentrations of each agonist were composed only of steady-state currents, but the responses to high concentrations were additionally composed of a rapid transient peak component except in the kainate-induced current. The currents induced by 10(-3)M N-methyl-D-aspartate in the external solution containing 0 mM Mg2+ and 10(-6)M glycine were reduced in amplitude when the external solution contained 1 mM Mg2+, and were abolished when the solution contained no glycine. The currents induced by a neurotransmitter candidate at retinogeniculate synapses, N-acetyl-aspartyl-glutamate, were markedly reduced in amplitude when the solution contained 1 mM Mg2+ or 10(-4)M DL-2-amino-5-phosphonovaleric acid. The current abolished in the Mg2+-containing, glycine-free solution (N-methyl-D-aspartate component) and the current remaining in the same solution (non-N-methyl-D-aspartate component) of the N-acetyl-aspartyl-glutamate response were both increased in a concentration-dependent manner, as the N-acetyl-aspartyl-glutamate concentration was increased. The current-voltage relationship of the currents induced by N-methyl-D-aspartate and N-acetyl-aspartyl-glutamate was characterized by Mg2+-dependent block at hyperpolarized potentials. The inward currents induced by 3 x 10(-4)M AMPA and 3 x 10(-4)M glutamate were markedly potentiated by 10(-4)M cyclothiazide, but the currents induced by 3 x 10(-4)M kainate and 10(-3)M N-acetyl-aspartyl-glutamate (non-N-methyl-D-aspartate component) were little affected. The currents induced by any agonist were not affected by 3 x 10(-4)g/ml concanavalin A. The current induced by 10(-4)M kainate was markedly suppressed by pretreatment with 10(-4)M AMPA or 10(-4)M glutamate, but only weakly by 10(-3)M N-acetyl-aspartylglutamate. The Ca2+ permeability (PCa/PCs) of the N-methyl-D-aspartate and non-N-methyl-D-aspartate receptors was 9.57 and 0.16, respectively. These results suggest that dorsal lateral geniculate nucleus relay neurons of the rat possessed both Ca2+-permeable N-methyl-D-aspartate receptors and less permeable non-N-methyl-D-aspartate (presumably AMPA) receptors, and that N-acetyl-aspartyl-glutamate mainly acts at N-methyl-D-aspartate receptors with a weak kainate-like action on non-N-methyl-D-aspartate receptors.

Animals↗

PubMed1999

Mu-opioid agonist-induced activation of G-protein-coupled inwardly rectifying potassium current in rat periaqueductal gray neurons.

The characteristics of the inwardly rectifying K+ current activated by a mu-type opioid agonist, D-Ala2,N-MePhe4,Gly5-ol-enkephalin (DAMGO), were examined in the acutely dissociated rat periaqueductal gray neurons using the nystatin-perforated and the conventional whole-cell recording modes under voltage-clamp conditions. DAMGO activated inward currents in a concentration- and voltage-dependent manner. The DAMGO-induced current was an inwardly rectifying K+ current (I(DAMGO)) which was sensitive to K+ channel blockers, quinine and Ba2+ but insensitive to Cs+ and tetraethylammonium. In the conventional whole-cell clamp mode, guanosine 5'-O-(2-thiodiphosphate) trilithium salt (GDPbetas, 0.4 mM) inhibited the amplitude of I(DAMGO) to 28% of that of the initial current. After the intracellular perfusion with guanosine 5'-O-(3-thiotriphosphate) tetralithium salt (GTPgammas, 0.4 mM) for 1 min, the first application of DAMGO irreversibly activated I(DAMGO). By the extracellular application of N-ethylmaleimide at a concentration of 50 microM for 2 min, I(DAMGO) was completely abolished. When a conventional whole-cell patch was made with a patch-pipette containing 1 microg/ml of pertussis toxin together with 1 mM of beta-nicotinamide adenine dinucleotide, I(DAMGO) gradually declined to about 41% of its initial amplitude. The extracellular application of second messenger modulators including protein kinase inhibitor (staurosporin), protein kinase A activators (forskolin, 3-isobutyl-l-methyl-xanthine and dibutyryladenosine 3'5'-cyclic monophosphate) and protein kinase C activators (phorbol-12-myristate-13-acetate and 1-oleoyl-2-acetyl-sn-glycerol) had no effect on I(DAMGO). These results suggest that (i) DAMGO-activated inwardly rectifying K+ current is mediated by pertussis toxin-sensitive guanine nucleotide binding proteins (G-proteins); (ii) the types of G protein involved in I(DAMGO) are Gi and/or Go; and (iii) the G-proteins exert their roles in I(DAMGO) without any mediation of the second messenger systems.

Animals↗

PubMed1999

Glutamate release from microglia via glutamate transporter is enhanced by amyloid-beta peptide.

In the present study, we found that amyloid-beta peptide enhanced glutamate release from primary cultured rat microglia via the Na+-dependent glutamate transporter, which was activated by extracellular K+. Glutamate transport current was measured by a conventional whole-cell patch recording mode under voltage-clamp conditions. With the pipette solution containing 10 mM glutamate and 100 mM Na+, an increase of the external K+ concentration from 0 to 10 mM evoked an outward current, resulting from co-extrusion of glutamate and Na+. The inward current, reflecting forward glutamate transport, was also activated by external glutamate. Both these reverse and forward glutamate transport currents were three-fold greater in microglia incubated with a relatively low concentration of amyloid-beta peptide (25-35) (5 microM) for four days. The glutamate-activated inward current was blocked by D,L-threo-beta-hydroxyaspartate in a dose-dependent manner (ranging from 0.001 to 1 mM), but not by a high concentration of kainate (1 mM). The glutamate concentration released from microglia upon high-K+ stimulation was also significantly increased (up to 170 microM) after treatment with amyloid-beta peptide (25-35). These results suggest that, at the pathological sites where extracellular K+ concentration may increase, the activation of microglia by amyloid-beta peptide causes an increase in extracellular glutamate concentration via reverse glutamate transporter, and therefore this mechanism may contribute to the pathogenesis of neuronal dysfunction and death in Alzheimer's disease.

ATP-Binding Cassette Transporters↗

PubMed1999

Calcium channels in the GABAergic presynaptic nerve terminals projecting to meynert neurons of the rat.

Effects of selective Ca2+ channel blockers on GABAergic inhibitory postsynaptic currents (IPSCs) were studied in the acutely dissociated rat nucleus basalis of Meynert (nBM) neurons attached with nerve endings, namely, the "synaptic bouton" preparation, and in the thin slices of nBM, using nystatin perforated and conventional whole-cell patch recording modes, respectively. In the synaptic bouton preparation, nicardipine (3 x 10(-6) M) and omega-conotoxin-MVIIC (3 x 10(-6) M) reduced the frequency of spontaneous postsynaptic currents by 37 and 22%, respectively, whereas omega-conotoxin-GVIA had no effect. After blockade of L- and P/Q-type Ca2+ channels, successive removal of Ca2+ from external solution had no significant effect on the residual spontaneous activities, indicating that N-, R-, and T-type Ca2+ channels are not involved in the spontaneous GABA release. Thapsigargin, but not ryanodine, increased the frequency of spontaneous IPSCs in both the synaptic bouton and slice preparations, suggesting the partial contribution of the intracellular Ca2+ storage site to the spontaneous GABA release. In contrast, omega-conotoxin-GVIA (3 x 10(-6) M) and omega-conotoxin-MVIIC (3 x 10(-6) M) suppressed the evoked IPSCs by 31 and 37%, respectively, but nicardipine produced no significant effect. The residual evoked currents were abolished in Ca2+-free external solution but not in the external solution containing 10(-5) M Ni2+, suggesting the involvement of N-, P/Q-, and R-type Ca2+ channels but not L- and T-type ones in the evoked IPSCs. Neither thapsigargin nor ryanodine had any significant effects on the evoked IPSCs. It was concluded that Ca2+ channel subtypes responsible for spontaneous transmitter release are different from those mediating the transmitter release evoked by nerve stimulation.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1999

Suppression of the nicotinic acetylcholine response in rat superior cervical ganglionic neurons by steroids.

The effects of various types of steroids on the nicotinic acetylcholine (ACh) receptor (nAChR)-mediated responses were investigated in superior cervical ganglionic neurons acutely dissociated from rats using nystatin perforated patch recording. ACh induced a peak followed by a gradual decrease in the inward current at a holding potential of -40 mV. Nicotine, but not muscarine, mimicked ACh. Hydrocortisone at a concentration of >10(-6) M reversibly suppressed both the peak and steady-state nicotine-induced currents (Inic) in a noncompetitive manner. The inhibition of Inic by hydrocortisone did not show any voltage dependency and persisted in the presence of either cyclic AMP modulators, forskolin and 3-isobutyl-1-methylxanthine, or a protein kinase A inhibitor, N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride (H-89). Beta-estradiol, androsterone, aldosterone, and 17alpha-estradiol mimicked hydrocortisone in its inhibitory action on ACh-induced currents (I(ACh)). The potency for the inhibitory actions on I(ACh) was as follows: androsterone > beta-estradiol > hydrocortisone > or = aldosterone = 17alpha-estradiol. Cholesterol had no effect on the I(ACh). In conclusion, the structural characteristics of a steroid are thus considered to be necessary to block nicotinic I(ACh) in rat superior cervical ganglionic cells, whereas the cholesterol side chain might disturb the inhibitory action of the steroid skeleton on nAChRs.

Acetylcholine↗

PubMed1999

Cholinergic systems in the nucleus of the solitary tract of rats.

The pharmacological and physiological properties of excitatory amino acid and ACh systems in the nucleus of the solitary tract (NTS) were studied in slices of rat brain stem by extracellular and intracellular recordings from neurons activated by solitary tract (ST) stimulation. These neurons were characterized as having several long dendrites with multiple varicosities. Synaptic activation of the medial NTS (mNTS) neurons by ST stimulation was mediated by non-N-methyl-D-aspartate (NMDA) glutamate (Glu) receptors, because the excitation was blocked by 6-cyano-7-nitro-quinoxaline-2,3-dione but not by NMDA, nicotinic, or muscarinic antagonists. Identified mNTS neurons were excited by iontophoresis of both Glu and ACh. The most sensitive region of the cell was on the dendrites approximately 100 micrometer from the cell body for both putative neurotransmitters. Nicotinic and/or muscarinic excitatory ACh responses were detected on the mNTS neurons. Our observations suggest that both types of ACh receptors may contribute to the attenuation of the baroreceptor reflex, but the functional correlation of this receptor profile remains to be determined.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1999

[Gramicidin perforated patch recording technique].

Cl- is one of the major ionic constituents of cells and extracellular spaces. Intracellular Cl- plays an important role in regulating the cell volume and pH, in both salt secretion and reabsorption, in membrane excitability, and G-protein-dependent intracellular signal transduction. GABA and glycine are the primary inhibitory neurotransmitters. Such agonist-stimulated responses are affected by the intracellular Cl- concentration ([Cl-]i). However, it was difficult to make an electrical recording of the physiological Cl- response from cells with native intracellular Cl- activity because of the limitations of present recording techniques using conventional glass-microelectrode, whole-cell patch and nystatin perforated patch recording modes. Recently, this difficulty was overcome by developing the gramicidin perforated patch recording mode in our laboratory. Gramicidin is a polypeptide antibiotic that forms pores in the cell membrane as well as nystatin but allows only monovalent cations to permeate the membrane, enabling both [Cl-]i and the second messenger system to remain undisturbed. Here, I would like to primarily focus on the GABA- and glycine-induced Cl- responses to in mammalian CNS neurons maintaining native cellular [Cl-]i under normal and pathological neuronal conditions by use of gramicidin perforated patch recording configuration. Age-related and developmental changes in neuronal [Cl-]i are also described in detail.

Animals↗

PubMed1999

Potassium-current oscillation of rat megakaryocytes: As a model system for drug evaluation (Review).

Megakaryocytes respond to externally applied agonists showing a periodic K+ current that reflects oscillation in cytoplasmic calcium concentration ([Ca2+]i). We have revealed several signal transducing factors that are involved in the K+ current oscillation of megakaryocytes. In this megakaryocyte system, it is relatively easy to determine what point of the signal transduction pathway a drug affects. In addition, as a progenitor cell, megakaryocytes resemble platelets which have important roles in many diseases. Therefore, this experimental system can be used for evaluation of new drugs.

Animals↗

PubMed1999

Na+,K+-ATPase activity in cultured bovine retinal pigment epithelium.

PURPOSE: To characterize electrogenic Na+,K+-ATPase activity in cultured bovine retinal pigment epithelium (RPE). METHODS: Cultured bovine RPE cells from passages 3 through 5 were dissociated enzymatically. Na+,K+-adenosine triphosphatase (ATPase)-activated currents (Ip) were measured by using a nystatin perforated-patch recording technique under voltage- clamp conditions. In the presence of suitable blockers for known voltage-dependent Na+, K+, and Ca2+ conductances, the Ip was activated in a concentration-dependent manner by adding K to the external solution. RESULTS: The median effective concentration (EC50) and Hill coefficient for external K+ concentration ([K+]o) were 1.06 mM and 2.55, respectively. The Ip showed no significant voltage dependency. A large outward shift of holding current was observed when [Na+]o, was removed. In the presence of [Na+]o, the addition of K+ to the external solution induced Ip, even when the internal solution did not contain Na+, suggesting the existence of a continuous Na+ influx across the plasma membrane in the presence of [Na+]o,. When Na+ was removed from the external and internal solutions, a transient Ip was observed, indicating that the transient Ip was activated by the intracellular residual Na+. The Ip was concentration-dependently suppressed by ouabain. The 50% inhibitory concentration (IC50) value and Hill coefficient for ouabain were 5.98 microM and 1.12, respectively. CONCLUSIONS: The present study is the first to reported the functional properties of electrogenic Na+,K+-ATPase activity in cultured bovine RPE.

Animals↗

PubMed1998

Effect of nilvadipine on the voltage-dependent Ca2+ channels in rat hippocampal CA1 pyramidal neurons.

Effects of nilvadipine on the low- and high-voltage activated Ca2+ currents (LVA and HVA ICa, respectively) were compared with other organic Ca2+ antagonists in acutely dissociated rat hippocampal CA1 pyramidal neurons. The inhibitory effects of nilvadipine, amlodipine and flunarizine on LVA ICa were concentration- and use-dependent. The apparent half-maximum inhibitory concentrations (IC50s) at every 1- and 30-s stimulation were 6.3x10-7 M and 1.8x10-6 M for flunarizine, 1.9x10-6 M and 7.6x10-6 M for nilvadipine, and 4.0x10-6 M and 8.0x10-6 M for amlodipine, respectively. Thus, the strength of the use-dependence was in the sequence of nilvadipine>flunarizine>amlodipine. Nilvadipine also inhibited the HVA ICa in a concentration-dependent manner with an IC50 of 1.5x10-7 M. The hippocampal CA1 neurons were observed to have five pharmacologically distinct HVA Ca2+ channel subtypes consisting of L-, N-, P-, Q- and R-types. Nilvadipine selectively inhibited the L-type Ca2+ channel current which comprised 34% of the total HVA ICa. On the other hand, amlodipine non-selectively inhibited the HVA Ca2+ channel subtypes. These results suggest that the inhibitory effect of nilvadipine on the neuronal Ca2+ influx through both LVA and HVA L-type Ca2+ channels, in combination with the cerebral vasodilatory action, may prevent neuronal damage during ischemia.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1998

Temperature-dependent effect of zolpidem on the GABAA receptor-mediated response at recombinant human GABAA receptor subtypes.

The effects of zolpidem on the two forms of recombinant human GABAA receptors (alpha1beta2gamma2s and alpha3beta2gamma2s) at different temperatures were functionally investigated, using the whole-cell patch recording configuration. In both forms, zolpidem potentiated the response to GABA in a concentration-dependent manner. At 16 degrees C, the apparent dissociation constant (KD) values for the alpha1beta2gamma2s and alpha3beta2gamma2s forms were 3.7 x 10(-8) and 5.6 x 10(-7) M, respectively. When the temperature was increased to 36 degrees C, the KD values for the alpha1beta2gamma2s and alpha3beta2gamma2s forms were 2.1 x 10(-7) and 1.5 x 10(-6) M, respectively. Although the affinity ratio was reduced from 15.1 to 7.1-fold the selectivity of zolpidem for the alpha1beta2gamma2s still remained at 36 degrees C.

GABA Agonists↗

PubMed1998

Characterization of electrogenic Na/K pump in rat neostriatal neurons.

The electrogenic Na/K pump current (Ip) was studied in the dissociated neostriatal neurons of the rat by using the nystatin-perforated patch recording mode. The Ip was activated by external K+ in a concentration-dependent manner with an EC50 of 0.7 mM at a holding potential (VH) of -40 mV. Other monovalent cations also caused Ip and the order of potency was Tl+>K+, Rb+>NH4+, Cs+>>>Li+. The Ip decreased with membrane hyperpolarization in an external solution containing 150 mM Na+, while the Ip did not show such voltage dependency without external Na+. Ouabain showed a steady-state inhibition of Ip in a concentration- and temperature-dependent manner at a VH of -40 mV. The IC50 values at 20 and 30 degrees C were 7.1 x 10(-6) and 1.3 x 10(-6) M, respectively. The decay of Ip after adding ouabain well fitted with a single exponential function. At a VH of -40 Mv, the association (k+1) and dissociation (k-1) rate constants estimated from the time constant of the current decay at 20 degrees C were 4.0 x10(2) s-1 M-1 and 6.3 x 10(-3) s-1, respectively. At 30 degrees C, k+1 increased to 2.8 x 10(3) s-1 M-1 while k-1 showed no such change with a value of 1.8 x 10(-3) s-1. A continuous Na+ influx was demonstrated by both the Na+-dependent leakage current and tetrodotoxin-sensitive Na+ current, which resulted in the continuous activation of the Na/K pump. It was thus concluded that the Na/K pump activity was well-maintained in the dissociated rat neostriatal neurons with distinct functional properties and that the activity of the pump was tightly connected with Na+ influxes.

Animals↗

PubMed1998

Inhibition of M-type K+ current by linopirdine, a neurotransmitter-release enhancer, in NG108-15 neuronal cells and rat cerebral neurons in culture.

The effect of linopirdine, a neurotransmitter-release enhancer, on the M-type K+-current, IK(M), was examined in NGPM1-27 cells, mouse neuroblastomaxrat glioma NG108-15 cells transformed to express m1-muscarinic acetylcholine (ACh) receptors, using the nystatin-perforated patch-recording mode under voltage-clamp conditions. The application of linopirdine induced the inward current associated with an inhibition of IK(M), which mimics an excitatory part of the ACh-induced responses in NGPM1-27 cells. The affinity of linopirdine for the inhibition of IK(M) was 24.7 microM in NGPM1-27 cells. In the presence of linopirdine, ACh failed to evoke a further inward current, but ACh still elicited an outward current, thus suggesting that the Ca2+-dependent K+ current is rather insensitive to linopirdine. Linopirdine also inhibited another voltage-gated potassium current (IK(V)) at the concentration of 72.3 microM. Finally, the inhibitory effect of linopirdine on IK(M) was confirmed in pyramidal neurons acutely dissociated from the rat cerebral cortex at 35.8 microM. The results suggest that linopirdine is thus considered to be an inhibitor of some type of K+ channels in both NGPM1-27 cells and the rat cerebral neurons.

Acetylcholine↗