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N Akaike

Publications and source records attributed to N Akaike.

At least 19 recordsLinked to original sources
PubMed1992

Effects of Ca2+ antagonists and antiepileptics on tetrodotoxin-sensitive Ca(2+)-conducting channels in isolated rat hippocampal CA1 neurons.

All Ca2+ antagonists blocked tetrodotoxin-sensitive Ca2+ current (TTX-ICa) more potently than Na+ current (INa). Phenytoin and MK-801, at concentrations which had no effect on INa, could block TTX-ICa concentration-dependently. Valproic acid and phenobarbital had no effect on both TTX-ICa and INa. In particular, flunarizine and phenytoin have more potent inhibitory effects on TTX-ICa than other test drugs. These results suggest that the abnormal excess-excitation of TTX-sensitive Ca(2+)-conducting channels may be one of the trigger factors generating epilepsy.

Animals↗
PubMed1992

A new type of Ca2+ channel blocker, NC-1100, inhibits the low- and high-threshold Ca2+ currents in the rat CNS neurons.

The effect of a new type of organic Ca2+ channel blocker, NC-1100 [(+/-)-1-(3,4-dimethoxyphenyl)-2-(4-diphenylmethylpiperazinyl)etha nol dihydrochloride], on both low- and high-threshold Ca2+ currents was studied in the whole-cell mode of the pyramidal neurons freshly dissociated from rat hippocampal CA1 region under voltage-clamp condition. The NC-1100 reversibly reduced the high-threshold Ca2+ current (HVA ICa) in a concentration-dependent manner without affecting the current-voltage relationship. The values of half-inhibition (IC50) were 1.3 x 10(-5) and 9.1 x 10(-6) M in external solution containing 10 and 2.5 mM Ca2+, respectively. The NC-1100 also decreased the low-threshold Ca2+ current (LVA ICa) in a concentration-dependent manner. The inhibitory potency was augmented by increasing the stimulation frequency and/or decreasing the extracellular Ca2+ concentration to a physiological range (2.5 mM). The IC50 value decreased to 7.7 x 10(-7) M in external solution containing 2.5 mM Ca2+ at a stimulation frequency of 1 Hz. The NC-1100 delayed the reactivation of LVA Ca2+ channel and enhanced voltage-dependently the steady-state inactivation, suggesting that this drug bound not only the resting LVA Ca2+ channel but also the inactivated one.

Animals↗
PubMed1992

Perforated-patch method reveals extracellular ATP-induced K+ conductance in dissociated rat nucleus solitarii neurons.

Extracellular adenosine 5'-triphosphate (ATP)-induced responses were investigated in acutely dissociated rat nucleus tractus solitarii (NTS) neurons, using nystatin perforated-patch and conventional whole-cell patch-clamp recordings. Extracellular application of ATP could evoke the inward currents with both methods. An additional outward current was observed only with the perforated patch method. The outward current resulted in an increase of K+ conductance which was activated by Ca2+ influx through the ATP receptor-channel complex.

Adenosine Triphosphate↗
PubMed1992

Penicillin-induced triphasic modulation of GABAA receptor-operated chloride current in frog sensory neuron.

Effects of penicillin-G (PCN) on GABA-evoked Cl- current (IGABA) were investigated in freshly dissociated frog sensory neurons by the use of the concentration-clamp technique combined with the suction-pipette method. Under conditions where the internal and external solutions allowed only Cl- permeability, PCN elicited triphasic modulation on IGABA, consisting of two modes of blockade on IGABA and a following rebound (rebound-like transient IGABA). Simultaneously applied PCN and GABA depressed IGABA immediately (phasic blockade), with the depressed IGABA slightly recovering in amplitude to achieve a stable level of blockade (tonic blockade). When a solution containing a mixture or PCN and GABA was quickly replaced by one containing GABA alone, a rebound-like transient Cl- current (IR) was evoked. Each component of the PCN actions on IGABA was PCN- and GABA-concentration-dependent. The reversal potential for each component of the PCN actions on IGABA was close to the chloride equilibrium potential (ECl) calculated using the Nernst equation. The current-voltage (I-V) relations for both the phasic and tonic blockade revealed inward rectification, while I-V curves for the control IGABA and the IR were outwardly rectified. The degree of IGABA-desensitization and the amplitude of the IR correlated well. The data suggest that partial removal of the GABAA receptor-desensitization may result in generation of the IR.

Animals↗
PubMed1992

Effects of Ca2+ antagonists and aminoglycoside antibiotics on Ca2+ current in isolated outer hair cells of guinea pig cochlea.

The effects of various Ca2+ antagonists and aminoglycoside antibiotics on the Ca2+ channel in isolated outer hair cells of the guinea pig were investigated using a whole-cell patch-clamp technique. The inhibitory action was in the order of La3+ much greater than Cd2+ much greater than Ni2+ greater than Co2+ for inorganic Ca2+ antagonists, and flunarizine = nicardipine greater than omega-conotoxin greater than methoxyverapamil = diltiazem much greater than amiloride for organic ones. Aminoglycoside antibiotics also had antagonistic effects on the Ca2+ channel.

Aminoglycosides↗
PubMed1992

Glycine response in isolated dorsal cochlear nucleus of C57BL/6J mouse.

Pharmacological properties of glycine (Gly)-induced Cl- current (ICl) in the dorsal cochlear nucleus (DCN) neurons acutely dissociated from C57BL/6J mouse were investigated in the whole-cell configuration of the patch-clamp technique. Gly-induced ICl increased in a sigmoidal manner with higher Gly concentrations. Strychnine blocked the Gly response competitively at low and non-competitively at high concentrations. Both glutamate (Glu) and N-methyl-D-aspartate (NMDA) responses were augmented by adding 10(-6) M Gly, at which concentration Gly did not induce any ICl. This facilitation was not affected by strychnine. Our results clearly show the existence of strychnine-sensitive and -insensitive glycine receptors in the DCN neurons.

Animals↗
PubMed1992

GABAergic synaptic current in dissociated nucleus basalis of Meynert neurons of the rat.

gamma-Aminobutyric acid (GABA)-mediated spontaneous inhibitory postsynaptic currents (IPSCs) were recorded from dissociated rat nucleus basalis of Meynert neurons which still had their synaptic boutons attached. The membrane currents were recorded by the whole-cell patch-clamp technique. Elevated extracellular K+ concentration and the addition of the calcium ionophore, A23187, enhanced the amplitude and frequency of spontaneous IPSCs. Ryanodine and Ca(2+)-free external solution containing EGTA or BAPTA markedly decreased the spontaneous IPSC activities. Spontaneous IPSC activities were reversibly reduced by baclofen and increased by phaclofen, indicating that the GABAB receptor regulates the release of GABA from nerve terminals and acts as a negative autoreceptor.

Animals↗
PubMed1992

Novel Ca2+ currents in mammalian CNS neurons.

Voltage-dependent Ca2+ currents (ICa) in neurons can be classified into T-, N- and L-types. In the CA1 pyramidal neurons freshly dissociated from rat hippocampus we found an additional tetrodotoxin (TTX)-sensitive Ca2+ current (termed 'TTX-ICa'). The TTX-ICa showed a heterogeneous distribution, preferentially in the dorsal site of CA1 region. Activation and inactivation processes of the TTX-ICa were highly potential-dependent, and the latter was fitted by a double exponential function. The TTX-ICa was activated at a threshold potential of about -55 mV and reached full activation at -30 mV. The steady-state inactivation of TTX-ICa could be fitted by a Boltzmann equation with a slope factor of 6.0 mV and a half-inactivation voltage of -72.5 mV. When the peak amplitudes of TTX-ICa were plotted as a function of extracellular Ca2+ concentration ([Ca2+]o), the current amplitude increased linearly without showing any saturation. The ratio of peak amplitude in the individual I-V relationships of Ca2+, Sr2+ and Ba2+ currents passing through the TTX-sensitive Ca(2+)-conducting channel was 1:0.33:0.05, although the current kinetics were much the same. TTX inhibited the TTX-ICa in time- and concentration-dependent manner without affecting the current kinetics. Lignocaine inhibited the TTX-ICa in a second in a concentration-dependent manner, with accelerating the inactivation process. The concentrations of half-inhibition (IC50) were 3.5 x 10(-9) M for TTX and 3.6 x 10(-4) M for lignocaine. Scorpion toxin prolonged the inactivation phase of TTX-ICa in a time- and concentration-dependent manner. In the toxin-treated neurons, both the slow time constant of inactivation (tau is) and its functional contribution to the total current increased with increasing the toxin concentration.

Animals↗
PubMed1992

Modulation of NMDA receptors by ACh in the central nervous system.

The effect of acetylcholine (ACh) on N-methyl-D-aspartate (NMDA)-induced responses in neurons acutely dissociated from the rat nucleus basalis of Meynert (nBM) was investigated with a conventional patch-clamp technique. The whole-cell recording revealed that NMDA-induced inward current (INMDA) consisted of transient peak and successive steady-state components. With outside-out recording, these components were shown to have the identical single channel conductance. Pretreating the nBM neurons with the low concentration of ACh suppressed the peak component of INMDA, whereas the high concentration potentiated it. The modulatory action of ACh was observed in nBM neurons dissociated from mature rats. The action was deficient, however, in those from immature animals. These results suggest that ACh is either an inhibitory or an excitatory modulator of glutamatergic transmission in the central nervous system.

Acetylcholine↗
PubMed1992

Regional variation of excitatory and inhibitory amino acid-induced responses in rat dissociated CNS neurons.

Regional differences in glutamate (Glu), aspartate (Asp), gamma-aminobutyric acid (GABA) and glycine (Gly) responses in CNS neurons were investigated by means of the whole-cell mode of the patch-clamp technique. The neurons were freshly dissociated from rat cortex, limbic system (hippocampal CA1 region), diencephalon (ventromedial hypothalamus), medulla (nucleus paragigantocellularis lateralis) and spinal cord (spinal dorsal horn). The current amplitudes induced by Glu and GABA did not show any regional differences whereas those of Asp- and Gly-induced responses were significantly different among CNS regions. The enhancement of Asp response by Gly was observed in all regions, and the facilitatory ratio did not differ among these regions. Even though the NMDA response in cortical neurons was significantly greater than that in spinal neurons, the ratios of NMDA response facilitation by Gly were also the same in both regions. When the current amplitudes induced by individual amino acids were estimated for the unit surface area of respective neurons (current density), the Glu, Asp and Gly responses showed regional heterogeneity whereas the GABA response did not.

Amino Acids↗
PubMed1992

5-HT response of rat hippocampal pyramidal cell bodies.

Serotonin (5-HT) responses of pyramidal neurones freshly dissociated from rat ventral hippocampal CA1 region were investigated by using nystatin-perforated whole-cell recording. These dissociated neurones lack most of the dendrites and axons. Application of nanomolar concentrations of 5-HT induced outward current with an increase of membrane conductance at a holding potential (VH) of -40 mV. The current was mimicked by alpha-methyl-5-HT (5-HT2 receptor family agonist), but not by 8-OH-DPAT (5-HT1 receptor family agonist). Ketanserin (5-HT2 receptor family antagonist) and spiperone (5-HT1A and 5-HT2 receptor family antagonist) blocked the current in a concentration dependent manner. These results suggests that 5-HT-induced outward current is mediated by the activation of 5-HT2 receptor family in the cell bodies of hippocampal pyramidal neurones.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1992

Non-competitive inhibition of GABAA responses by a new class of quinolones and non-steroidal anti-inflammatories in dissociated frog sensory neurones.

1. The interaction of a new class of quinolone antimicrobials (new quinolones) and non-steroidal anti-inflammatory agents (NSAIDs) with the GABAA receptor-Cl- channel complex was investigated in frog sensory neurones by use of the internal perfusion and 'concentration clamp' techniques. 2. The new quinolones and the NSAIDs (both 10(-6)-10(-5) M) had little effect on the GABA-induced chloride current (ICI) when applied separately. At a concentration of 10(-4) M the new quinolones, and to a lesser degree the NSAIDs, produced some suppression of the GABA response. 3. The co-administration of new quinolones and some NSAIDs (10(-6)-10(-14) M) resulted in a marked suppression of the GABA response. The size of this inhibition was dependent on the concentration of either the new quinolone or the NSAID tested. The inhibitory potency of new quinolones in combination with 4-biphenylacetic acid (BPAA) was in rank order norfloxacin (NFLX) much greater than enoxacin (ENX) greater than ciprofloxancin (CPFX) much greater than ofloxacin (OFLX), and that of NSAIDs in combination with ENX was BPAA much greater than indomethacin = ketoprofen greater than naproxen greater than ibuprofen greater than pranoprofen. Diclofenac, piroxicam and acetaminophen did not affect GABA responses in the presence of ENX. 4. In the presence of ENX or BPAA, there was a small shift to the right of the concentration-response curve for GABA without any effect on the maximum response. However, the co-administration of these drugs suppressed the maximum of the GABA concentration-response curve, indicating a non-competitive inhibition, for which no voltage-dependency was observed.5. Simultaneous administration of ENX and BPAA also suppressed pentobarbitone (PB)-gated Icl. On the other hand, both PB and phenobarbitone reversed the inhibition of GABA-induced Ic, by coadministration of ENX and BPAA.6. The effect on GABAA responses of co-administration of new quinolones and NSAIDs was not via an interaction with benzodiazepine receptors coupled to the GABAA receptor, since this effect was not reversed by Rol5-1788 or diazepam.7. It is concluded that the co-administration of new quinolones and some of the NSAIDs inhibit GABAergic transmission, and could result in convulsions.

Animals↗
PubMed1992

Penicillin-induced potentiation of glycine receptor-operated chloride current in rat ventro-medial hypothalamic neurones.

1. Effects of penicillin G (PCN) on glycine (Gly)-evoked Cl- current (IGly) were investigated in acutely dissociated rat ventro-medial hypothalamic (VMH) neurones by the whole cell mode of patch clamp technique. 2. When PCN was applied simultaneously with Gly, PCN depressed IGly like a Cl- channel blocker. 3. The PCN-induced blocking action was clearly observed at a low PCN concentration (30 u), while the maximal blockade was achieved by 600 u (units per 10 ml) PCN. 4. When tested solution containing both PCN and Gly was quickly substituted with one containing Gly only, a new rebound-like transient current (I(T)) which also passed through Cl- channel, was elicited. 5. The peak amplitude of I(T) induced by PCN at concentrations higher than 100 u was greater than that induced by glycine alone. We termed this phenomenon PCN-induced potentiation of IGly. In all cells tested, PCN potentiated IGly. 6. At a lower PCN concentration below 30 u, I(T) generation was not clear in the presence of 10(-5) M gamma-aminobutyric acid. With PCN a higher concentration than 300 u, I(T) amplitude was greater than that of the original peak IGly. This was observed in 18 neurones out of 21. The maximal amplitude of the I(T) was achieved with 600 u PCN.

Animals↗
PubMed1992

Strychnine-induced potassium current in CA1 pyramidal neurones of the rat hippocampus.

1. Direct actions of strychnine (Str) and brucine (Bru) on the dissociated hippocampal CA1 neurones of the rat have been investigated with the whole-cell mode of the patch-clamp technique. 2. At a holding potential (VH) of -20 mV, both Str and Bru elicited outward current at concentrations over 10(-5) M. The reversal potential of Str-induced current (EStr) was -77.8 mV, which was close to the K+ equilibrium potential (EK = -80.3 mV). The change in EStr for a ten fold change in extracellular K+ concentration was 58 mV, indicating that the membrane behaves like a K+ electrode in the presence of Str. 3. The concentration-response curves for Str and Bru were bell-shaped, and nearly maximum response occurred at 10(-4) M for Str and 3 x 10(-4) M for Bru. The maximum current amplitude induced by Bru was about 80% of that induced by Str. A transient 'hump' current appeared immediately after the wash-out of external solutions containing Str and Bru at concentrations higher than 10(-4) and 3 x 10(-4) M, respectively. 4. The Str-induced current (IStr) was antagonized by K+ channel blockers such as Ba2+, tetraethylammonium (TEA)-chloride, and 4-aminopyridine (4-AP) in a concentration-dependent manner. IStr was insensitive to glibenclamide, a blocker of ATP-sensitive K+ channels. 5. Internal perfusion with 10 mM BAPTA did not affect the Str-induced IK. Depletion of the intracellular Ca2+ store by caffeine had no effect, indicating that intracellular Ca2+ does not mediate the Str-induced activation of K+ conductance.6. Both guanosine-5'-0-3-thiotriphosphate (GTPyS) and guanosine-5'-O-thiodiphosphate (GDPPS) suppressed the Str-induced IK, the former action appearing more rapidly than the latter. The results suggest that the GTP binding proteins are involved in this Str response.7. When neurones were loaded with cholera toxin (CTX) or pertussis toxin (PTX) through a patch pipette, PTX suppressed the Str response whereas CTX did not, suggesting that G, and/or Go might be involved in the Str-induced IK.

Animals↗
PubMed1992

Blockade of 5-HT3 receptor-mediated currents in dissociated frog sensory neurones by benzoxazine derivative, Y-25130.

1. The effect of Y-25130, ((+-)-N-(1-azabicyclo[2.2.2]oct-3-yl)-6-chloro-4-methyl-3-oxo-3,4-dih ydr o- 2H-1,4-benzoxazine-8-carboxamide hydrochloride), a high affinity 5-hydroxytryptamine3 (5-HT3) receptor ligand, was examined on the 5-HT-induced response in dissociated frog dorsal root ganglion (DRG) neurones by use of the extremely rapid concentration-jump ('concentration-clamp') and the conventional whole-cell patch-clamp techniques. 2. 5-HT induced a rapid transient inward current associated with an increase in membrane conductance at a holding potential of -70 mV. The current amplitude increased sigmoidally as 5-HT concentration increased. The half-maximum value (Ka) and the Hill coefficient estimated from the concentration-response curve were 1.7 x 10(-5) M and 1.7, respectively. 3. The current-voltage (I-V) relationship of 5-HT-induced current (I5-HT) showed inward rectification at potentials more positive than -40 mV. The reversal potential (E5-HT) was -11 mV. The E5-HT value was unaffected by total replacement of intracellular K+ by Cs+, indicating that the 5-HT-gated channels might be large cation channels. 4. Both the activation and inactivation phases of I5-HT were single exponentials. The time constants of activation and inactivation (tau a and tau i) decreased with increasing 5-HT concentration. 5. The 5-HT response was mimicked by a selective 5-HT3 receptor agonist, 2-methyl-5-HT, but the maximum response induced was approximately 25% that of 5-HT. The 5-HT response was reversibly antagonized by the 5-HT3 receptor antagonists, ICS 205-930, metoclopramide and Y-25130, but not by a 5-HTIA receptor antagonist, spiperone, and a 5-HT2 receptor antagonist, ketanserin. The half-inhibition concentrations (IC50) were 4.9 x 10-10 M for Y-25130, 4.8 x 10-10 M for ICS 205-930 and 8.6 x 10-9 M for metoclopramide.6. Y-25130 (5 x 10-10 M) caused a rightward shift of the concentration-response curve for 5-HT while decreasing the maximum response.7. The results suggest that Y-25130 is a potent antagonist of the 5-HT3 receptor-channel complex.

Animals↗
PubMed1992

Proton-induced sodium current in freshly dissociated hypothalamic neurones of the rat.

1. The proton-gated current was investigated in freshly dissociated ventromedial hypothalamic (VMH) neurones from 4-week-old Wistar rats, under whole-cell configuration by the use of the 'concentration-clamp' technique which combines intracellular perfusion with the rapid exchange of external solution within 1-2 ms under a single-electrode voltage-clamp condition. 2. The proton-gated current increased in a sigmoidal fashion as extracellular pH (pHo) decreased. In external solution containing 2 mM-Ca2+, the threshold of current activation was at pHo 6.5, and the maximum response appeared at pHo 4.1-3.9. The dissociation constant (Kd) and Hill coefficient were 10(-4.9) M (pHo = 4.9) and 1.5 respectively. 3. Decreasing extracellular Na+ concentration reduced the proton-gated current. The current reversed direction at the Na+ equilibrium potential (ENa), indicating that it was carried by Na+. 4. The activation phase kinetics of proton-induced current was single exponential. The time constant of activation (tau a) did not have a potential dependence but decreased slightly by decreasing pHo. The inactivation phase kinetics was two-exponential. The time constant of inactivation (tau i) consisted of fast and slow components (tau if and tau is, respectively). Like tau a, both tau if and tau is did not have any potential dependence, but they slightly increased with decreasing pHo. 5. The steady-state inactivation curve, constructed by decreasing pHo from various conditioning pHos to 4.1, revealed that the proton-induced current had a half-maximum inactivation at pHo 6.2. 6. The proton-induced current was suppressed as the extracellular Ca2+ concentration ([Ca2+]o) increased from almost free (0.01 mM) to 80 mM. Increasing [Ca2+]o increased tau a, but slightly decreased both tau if and tau is. 7. Recovery of proton-induced current from complete inactivation of proton-induced current depended on the degree of pHo change. A bigger change in pHo induced faster recovery than a smaller change. 8. External divalent cations inhibited the proton-induced current, and the inhibitory potency was in the order of Mn2+ greater than Co2+ greater than Ca2+ greater than Sr2+ greater than Ba2+ greater than Mg2+. 9. Tetrodotoxin (TTX) at relatively low concentration (less than 10(-7) M) did not inhibit the peak amplitude of the proton-induced current, but at a higher concentration (10(-6) M) it slightly inhibited the peak amplitude of the current and accelerated the inactivation process. Scorpion toxin markedly increased the peak amplitude of the proton-induced current and prolonged the inactivation phase. The tau is was also increased by scorpion toxin in a concentration-dependent manner. Veratridine had no effect on the proton-induced current. 10. The membrane properties of the proton-operated channel were similar to those of the voltage-gated Na+ channel rather than the Ca2+ channel.

Animals↗
PubMed1992

Direct modulation of GABAA receptor by intracellular ATP in dissociated nucleus tractus solitarii neurones of rat.

1. Effect of intracellular ATP on Cl- current (ICl) mediated by the GABA (gamma-aminobutyric acid) receptor subtype, GABAA, was studied in dissociated nucleus tractus solitarii (NTS) neurones using the whole-cell mode of patch clamp. A concentration-jump technique termed 'Y tube' was used to rapidly apply agents externally. Dissociated neurones were obtained from 1- to 3-week-old rats. 2. When the patch-pipette solution contained 2 mM-ATP, the amplitude of ICl elicited by 10(-5) M-GABA did not show any time-dependent decrease (apparent run-down), for more than 60 min after the initial recording. In the presence of ATP, the half-maximum concentration (KD) and Hill coefficient calculated from the GABA concentration-response curve were 9.12 microM and 1.47, respectively. 3. In the absence of intracellular ATP, the amplitude of GABA-induced ICl decreased with time. The relative peak amplitudes after 20 and 60 min from the initial recording were 0.40 +/- 0.09 (n = 11) and 0.16 +/- 0.05 (n = 8) with respect to the initial response. 4. Removal of Mg2+ from the internal solution induced run-down of the GABA response even in the presence of 2 mM-intracellular ATP, suggesting that both intracellular ATP (2 mM or more) and Mg2+ are necessary to prevent run-down of the GABA response. 5. Activation of dephosphorylation processes by alkaline phosphatase (100-200 microM) did not affect the GABA response in neurones perfused with internal solution containing 2 mM-ATP and 3 mM-Mg2+. Blocking the dephosphorylation process by okadaic acid, a phosphatase inhibitor, did not prevent the run-down of the GABA response. 6. Calcium influxes passing through both the voltage-dependent L-type Ca2+ channel and the glutamate receptor-operated cation channel did not affect ICl induced by GABA. 7. GABA-induced ICl was also maintained by adding 2 mM-ADP or ATP gamma S (adenosine-5'-O-3-thiotriphosphate) to the internal solution containing Mg2+. Addition of 2 mM-adenosine, AMP, cyclic AMP, AMP-PNP (adenylimido-diphosphate) or ADP beta S (adenosine-5'-O-2-thiodiphosphate) to the internal solution did not prevent the run-down of the GABA response even in the presence of 3 mM-intracellular Mg2+. Based on the chemical specificity of these ATP analogues, it is suggested that there is an ATP-sensitive binding site (ATP receptor) in the cytoplasmic side of the cell membrane.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenosine Triphosphate↗