Biomedical subjects

N Akaike

Publications and source records attributed to N Akaike.

At least 325 records · Page 18Linked to original sources

PubMed1988

Effects of Ca2+ on the transient outward current of single isolated Helix central neurones.

1. Single Helix neurones were studied under voltage-clamp conditions with internal perfusion in order to examine the contribution of internal and external Ca2+ and the effects of 4-aminopyridine (4-AP) on the transient outward current (IA). 2. In Na+ -free snail Ringer, replacement of external Ca2+ [( Ca2+]o) with equimolar Co2+ reduced the maximum amplitude of IA and induced a shift of the steady-state part of the IA inactivation curve (I-V curve), in a positive direction along the voltage axis when the neurone was internally perfused with K-aspartate. 3. In Ca2+ -free solutions, precipitation or chelation of internal Ca2+ [( Ca2+]i) by internal perfusion with KF or EGTA shifted the curve in a more negative direction without affecting the maximum amplitude of IA. Thus, the kinetics of IA are dependent not only upon [Ca2+]o, as previously suggested, but also upon [Ca2+]i. 4. In the presence of 4-AP the I-V curve for IA shifted in a hyperpolarizing direction and the maximal amplitude was reduced when the neurone was internally perfused with K-aspartate. However, 4-AP had little or no effect on the I-V relationship of IA when the neurone was internally perfused with the Ca2+ precipitating or chelating agent, KF or EGTA. These results suggest that the actions of 4-AP on IA are at least partly dependent upon [Ca2+]i.

4-Aminopyridine↗

PubMed1988

The effect of temperature on the GABA-induced chloride current in isolated sensory neurones of the frog.

1. The effect of temperature on the kinetics of the activation and inactivation phases of gamma-aminobutyric acid (GABA)-induced Cl- current (ICl) was examined in frog isolated sensory neurones. 2. The peak ICl was reversibly reduced on changing the temperature and temperature-dependent coefficients were shown to exist, with the highest Q10 (1.58) occurring between 5-15 degrees C. 3. At both room temperature (20 degrees C) and 10 degrees C, the GABA dose-response curve was sigmoidal with a Hill coefficient of 2 and half-maximal responses to GABA, Kd, of 1.3 x 10(-5)M and 1.1 x 10(-5)M, respectively. Thus, indicating no change in the binding affinity of GABA when the temperature was decreased. 4. At GABA concentrations greater than 10(-5)M, both the activation and inactivation phases of the GABA-induced ICl consisted of double exponentials, fast and slow components respectively, in the temperature range of 10 to 30 degrees C. 5. The fast (tau af) and slow (tau as) activation time constants decreased with an increase in temperature and increased with a reduction in temperature. With an increased temperature, the reduction in peak ICl was due to a reduction in the slow time constant with no significant change in the fast time constant. 6. Both the fast (tau if) and slow (tau is) inactivation time constants were also increased by cooling to 10 degrees C; heating to 30 degrees C had little effect. 7. The concentration-dependence (10(-5) to 10(-3)M) of the slow activation (tau as) and inactivation (tau is) time constants was unaltered by the change in temperature. Similarly, the lack of concentration-dependence shown by the fast activation (tau af) and inactivation (tau if) time constants was unaltered by the temperature change. 8. From recordings made with 'inside-out' patches, the probability of opening of the GABA-induced Cl- channels showed a marked increase with cooling to 10 degrees C compared to room temperature (20 degrees C), with no change in channel conductance. 9. The change in the GABA-induced ICl at different temperatures is, therefore, not due to changes in binding but to subsequent channel activation. Possible mechanisms whereby this occurs are discussed.

Animals↗

PubMed1988

Kinetic and pharmacological properties of the GABA-induced chloride current in Aplysia neurones: a 'concentration clamp' study.

1. gamma-Aminobutyric acid (GABA) was applied by the 'concentration clamp' technique to isolated neurones of Aplysia. GABA induced a chloride current (ICl) due to activation of a single class of chloride-channel. 2. The concentration-response curve for the peak ICl gave an apparent dissociation constant of 6.4 X 10(-5) M and a Hill coefficient of 0.88. The current-voltage relationship was linear in the voltage range examined (-40 to +10 mV). 3. The activation phase of the ICl could be fitted to a single exponential function and desensitization followed the sum of two exponential functions. The time constants of activation and desensitization decreased with increasing concentrations of GABA but were voltage-independent. The recovery process from desensitization also followed the sum of two exponential functions. 4. As for the rate-limiting step of the channel activation, the hyperbolic relationship between the activation rate and GABA concentration showed that the rapid binding assumption holds, suggesting that the isomerization step is rate-limiting. The apparent channel closing rate constant was estimated to be 10 s-1 from the ordinate intercept of the linear part of the above relationship at lower concentrations. 5. Muscimol and beta-alanine induced a ICl, which cross-desensitized with that evoked by GABA. The GABA-ICl was not enhanced by diazepam (10(-6) M) or alpha-chloralose (10(-3) M), in fact depressant effects were evident. 6. Pentobarbitone decreased the GABA-ICl non-competitively without altering activation or desensitization kinetics. The concentration-inhibition curve gave a KD value of 8.9 x 10(-5) M and a Hill coefficient of 1.0. 7. These results suggest that GABA activates a single class of Cl channel in Aplysia neurones, which have one binding site for the agonist. The GABA receptor-Cl channel complex in Aplysia is pharmacologically and perhaps structurally different from that in vertebrates.

Animals↗

PubMed1988

Ca-activated K channel in cultured smooth muscle cells of rat aortic media.

Electrical and pharmacological properties of the single Ca-activated K channel in cultured smooth muscle cells (SMC) of the rat aorta were studied with the patch-clamp technique. The Ca-activated K channel had a slope conductance (gamma K) of 135 +/- 2 pS (mean +/- SE; n = 5) in symmetrical 142 mM K solutions. The reversal potentials show a 56-mV change for a 10-fold change in the external K concentration. Probability of the channel opening increased when the intracellular Ca concentration ([Ca]i) was increased over 10(-7) M or the membrane was depolarized. The channel was blocked by either external tetraethylammonium (TEA, 10-30 mM) or by internal Ba (1-5 mM). Channel activities were characterized by burst-like openings. Open-time histogram was fitted with a single exponential (tau = 1.3 ms at +10 mV and 10(-7) M [Ca]i), whereas the closed-time histogram was fitted with two exponentials (tau 1 = 0.7 ms and tau 2 = 111 ms). The permeability ratio for monovalent cations calculated with the Goldman-Hodgkin-Katz equation was K:Rb:Na = 1:0.7: less than 0.01. We conclude from these observations that the Ca-activated K channel in cultured SMC of the rat aorta is characterized by a middle size gamma K, activation by [Ca]i increase and depolarization, relatively low sensitivity to TEA, and high selectivity for K ions.

Animals↗

PubMed1988

Cyclic AMP modulates Ca-activated K channel in cultured smooth muscle cells of rat aortas.

Effects of adenosine 3',5'-cyclic monophosphate (cAMP) on single Ca-activated K current (IK(Ca)) in cultured smooth muscle cells of the rat aorta were investigated with the patch-clamp technique. In cell-attached patch configurations, extracellular application of isoproterenol (10(-5) M) increased the Ca-activated K currents. The increase in the currents was due to an increase in the probability of channel openings (Po). Neither unit conductance nor the maximum number of the channel in the patch was affected by the drug. The effects were inhibited by adding propranolol (10(-6) M). The extracellular application of forskolin (10(-5) M) or dibutyryl cAMP (10(-4) M) mimicked the effects of isoproterenol. In inside-out patch configurations, activated cAMP-dependent protein kinase (A kinase) in the bathing solution increased the sensitivity of the Ca-activated K channels to intracellular free calcium concentration ([Ca]i) and enhanced Po. Kinetic analyses of the IK(Ca) showed that cAMP-dependent phosphorylation of the Ca-activated K channels significantly reduced the mean closed time between bursting openings. We conclude from these observations that the Ca-activated K channels in aortic cells may increase Po through cAMP-dependent phosphorylation.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1987

Inhibition of Na+-dependent GABA uptake in isolated frog sensory nerve cell bodies by extra- and intracellular Li+.

The effect of Li+ on the Na+-dependent gamma-aminobutyric acid (GABA) uptake was investigated by examining the effects of external and internal Li+ ([Li]o and [Li]i) on GABA-gated Cl- current (ICl) in the frog dorsal root ganglion cells using the suction pipette technique which allows the internal perfusion under voltage-clamp condition. The suppression of GABA responses in the presence of external Na+ ([Na]o) was larger at lower than at higher GABA concentration. Replacement of [Na]o with Li+ completely removed the Na+ suppression, and GABA dose-response curve in Li+ external solution agreed well with that in Na+-free (Tris+) external solution. Either increasing [Li]i) or internal Na+ ([Na]i) at a constant [Na]o equally reduced the Na+-dependent suppression of GABA-gated ICl. The results indicate that both the [Li]o and [Li]i remove in different manner the [Na]o-dependent suppression of GABA-induced ICl:i.e. the [Li]o acts as a blocker of Na+-GABA co-transport mechanism while the increase of [Li]i decreases the Na+ electrochemical potential gradient across the soma membrane as well as [Na]i does.

Animals↗

PubMed1987

Synthetic omega-conotoxin: a potent calcium channel blocking neurotoxin.

The Ca2+ channel blocking action of synthetic omega-conotoxin (omega CTX) was studied on isolated frog dorsal root ganglion neurons using a 'concentration clamp' technique which enabled internal perfusion and rapid external solution change. At 100 nM, omega CTX showed a time-dependent depression of Ca2+ current (ICa). At higher concentrations, omega CTX exhibited a dose-dependent depression of ICa amplitude without changing the current-voltage relationship. Increases in external Ca2+ concentration partly overcame the inhibitory action of omega CTX on the ICa amplitude. At 10 microM omega CTX totally blocked ICa without effect on the Na+ current. It was likely that omega CTX had high selectivity for the Ca2+ channel.

Animals↗

PubMed1987

Spider toxin blocks excitatory amino acid responses in isolated hippocampal pyramidal neurons.

Using the 'concentration-clamp' technique we have investigated the action of Joro spider toxin (JSTX), as a specific blocker of glutamate receptor, on freshly isolated rat hippocampal pyramidal neurons. The neurons showed prominent responses to L-glutamate (L-Glu), quisqualate (QA) and kainate (KA) and JSTX blocked completely the both responses. The blocking action of toxin was dose-dependent at the concentrations of toxin between 4.8 X 10(-12) and 4.8 X 10(-8) M, and was remarkably similar in cell to cell trials. The kinetics of blockade was revealed by applying the toxin to the non-desensitizing KA response. We suggest that the JSTX spider toxin may be a valuable tool in ligand binding studies of QA/KA receptors in the central nervous system.

Animals↗

PubMed1987

D-600 blocks open Ca2+ channels more profoundly than closed ones.

The open channel blocking action of the Ca2+ antagonists, D-600 and Cd2+, was investigated in single neurons isolated by enzymatic treatment from dorsal root ganglia of frog. Using a 'concentration clamp' (jump) technique Ca2+ antagonists were applied to the preparation just before (less than 100 ms) or during a depolarizing step which induced the maximal peak amplitude of Ca2+ current (ICa). The inhibitory action of D-600 was more pronounced when applied during a depolarizing step than when applied just before a depolarizing step. In contrast, Cd2+ was equally inhibitory, whether applied before or during the depolarization. Thus, D-600 affects open channels more quickly or more profoundly than closed ones, while Cd2+ acts equally on both open and closed Ca2+ channels.

Animals↗

PubMed1987

Active sodium-potassium transports in skeletal muscles of deoxycorticosterone hypertensive rats.

CNS-induced suppressions of active Na+, K+ transport was investigated in both 'tonic' muscles, soleus (SOL), and 'twitch' muscle, extensor digitorum longus (EDL) of deoxycorticosterone acetate (DOCA) hypertensive rats. There was a marked K+ loss and Na+ accumulation in the skeletal and smooth muscles of DOCA hypertensive rats. The cellular K+ loss was in the order of SOL greater than EDL greater than diaphragm greater than intestine greater than aorta. However, liver, kidney and CNS organs such as cerebrum, cerebellum and medulla oblongata were spared from this K+ fall. Sciatic nerve sectioning or cervical transection activated the active Na+, K+ transport in SOL during DOCA hypertension but inhibited further the pump activity in EDL. The application of tetrodotoxin on the sciatic nerve also activated the Na+, K+ transport in SOL but inhibited the transport in EDL. The facilitatory effect of denervation on the pump activity in SOL was abolished by pretreatment with ouabain. Injection of curare had no effect on Na+ and K+ contents in both SOL and EDL. These results indicate that the CNS is involved differently on the neural regulations of the active Na+, K+ transport systems in SOL and EDL of DOCA hypertensive rats.

Animals↗

PubMed1987

4-aminopyridine activates a cholinergic chloride conductance in isolated Helix neurons.

Isolated Helix neurons were internally perfused under voltage-clamp, with drugs applied by the 'concentration clamp' which allowed an extremely rapid switching of the external solution. 4-Aminopyridine (4-AP) at more than 3 X 10(-4) M induced a chloride current (ICl) which could be blocked by D-tubocurarine. Acetylcholine (ACh) also induced ICl which was similarly blocked. Simultaneous application of 4-AP and ACh resulted in potentiation of ICl over the sum of respective currents. When preceded by either one, the other ICl was reduced in size, showing cross-desensitization. These data indicate that 4-AP activates ICl by binding to the cholinergic receptors, and reduces ACh-induced ICl by desensitizing a common receptor-channel complex.

4-Aminopyridine↗

PubMed1987

Different actions of intracellular free calcium on resting and GABA-gated chloride conductances.

The effects of voltage-dependent Ca2+ current (ICa) on resting and gamma-aminobutyric acid (GABA)-gated Cl- conductances were studied in isolated frog sensory neurons. The amplitude of GABA-gated Cl- current (ICl) was greatly suppressed by a preceding ICa. The GABA dose-response curve was shifted to the right without changing the maximum response by increasing [Ca2+]i. An ICa-activated ICl was observed as an inward tail current on the 'off' phase of ICa. This tail current was easily saturated by increasing the amount of Ca2+ influx.

Animals↗

PubMed1987

Multiple mechanisms of antagonism of gamma-aminobutyric acid (GABA) responses.

Gamma-aminobutyric acid (GABA) is one of the most important neurotransmitters in the brain. In an effort to understand the operation of the GABA receptor-ionophore complex, the antagonism of GABA responses by four substances was studied in bullfrog dorsal root ganglion cells by concentration-clamp and internal-perfusion techniques. Two antagonists (bicuculline and Zn2+) were competitive; two (picrotoxin and penicillin) were noncompetitive. However, significant changes in the kinetics of activation and inactivation were produced by the antagonists, including those that were competitive. The causes of these changes may be important clues to the structure and operation of the GABA receptor-ionophore complex.

Animals↗

PubMed1987

Piperidine: a microelectrophoretic study in the mammalian brain.

Using unit recording and electrophoretic techniques, the action of piperidine on unit activity of the brain of the rat was studied. Piperidine excited 31%, and inhibited 4% of cortical cells tested. In the hippocampus and caudate nucleus, piperidine excited larger proportions of the cells tested. The actions of piperidine were blocked by tetraethylammonium but not by scopolamine.

Acetylcholine↗

PubMed1987

Selective effects of enzyme treatment used for dissociating single cells on their GABA-receptor activities.

Frog dorsal root ganglion (DRG) cells treated with collagenase and trypsin were dissociated mechanically. The characteristics of dose-response curves of GABA-induced Cl- currents (ICl) were mainly studied using the "concentration-clamp" technique, by which the external solution around isolated single neurons was rapidly changed within a few milliseconds. The saturating maximum currents induced by various concentrations of GABA normalized to the peak ICl induced by 10(-5) M GABA on the same neuron varied from 1.5 to 5.5 among the neurons, though the absolute maximum currents varied little. The Hill coefficients (n = 1.92 +/- 0.09, mean +/- S.D.) also varied little on the neurons. The time constants of the desensitization phase of fully activated GABA responses were not so markedly different among the neurons. The present results suggested a possibility that the enzyme treatment produced a selective inactivation of alpha 1 unit, a high affinity subtype of the GABA-receptor.

Animals↗

PubMed1987

Comparison of red and white muscle wet weight changed by age, denervation and morbid states.

[Na]i, [K]i and wet weight of the extensor digitrum longus (EDL) and soleus (SOL) muscles of 9- and 52-week-old rats were measured for 7 days after sectioning of the sciatic nerve. The changes in wet weight of the EDL and SOL muscles of rats over 52 weeks and those of morbid state rats were also measured. There was no significant difference in wet weights between the EDL and SOL muscles in infant rats, but the EDL muscle became much heavier than the SOL muscle with aging. The decrease in rate of growth of wet weight of the EDL and SOL muscles caused by denervation, was greater in young rats than in mature rats. In addition, the rate of decrease was greater in the SOL muscles than in the EDL muscles in both young and mature rats. The [Na]i increased while [K]i was decreased by denervation, and the net Na+ increase and the net K+ loss were greater in young rats than in mature rats. The changing rate was more remarkable in the EDL muscles than in the SOL muscles throughout the aging process. During DOCA treatment over 4 weeks, the decrease of muscle wet weight was greater in the EDL muscles. The mechanisms which serve to maintain normal muscle wet weight in the SOL muscle after denervation or treatment with DOCA, were discussed.

Aging↗

PubMed1987

Antagonists of GABA responses, studied using internally perfused frog dorsal root ganglion neurons.

Responses of frog dorsal root ganglion neurons to GABA were studied under conditions of internal perfusion. Conductances to Na, Ca and K were pharmacologically blocked, C1 concentrations were maintained equal on both sides of the membrane and a small holding potential was used. Under these conditions GABA-induced C1 currents could be studied in isolation without shifts in EC1 occurring after GABA application. GABA currents were blocked by a variety of agents. The blockade by bicuculline and Zn was competitive, while that to penicillin was competitive at low concentrations (6 x 10(-5) M) and non-competitive at high concentrations (3 x 10(-4) M). Picrotoxin was non-competitive at all concentrations studied. The time course of the GABA-induced currents was changed in the presence of antagonists, including those that were competitive. These actions appear to be due to a change in the rates of receptor desensitization rather than shifts in EC1. Pretreatment with antagonists increased the degree of inhibition only for picrotoxin as compared to simultaneous application of GABA plus antagonist. The voltage dependence of the GABA response was altered by penicillin but not by other antagonists. GABA responses on frog dorsal root ganglion cell were also depressed by a variety of other metal ions (Cd, Ni, Cu, Co, Mn) and other drugs (strychnine, curare, 4-acetamide, 4'-isothiocyano-stilbene-2,2'-dilsulfonic acid disodium salt, 4,4'-diisothiocyano-stilbene-2,2'-dilsulfonic acid disodium salt trihydrate, bemegride and folic acid). In this preparation bicuculline and the heavy metal ions appear to block at or very near to the agonist binding site, while penicillin probably blocks the ion channel. The non-competitive action of picrotoxin appears not to be channel blockade, but to be due to a slowly equilibrating action at a site different from either the agonist binding site or the channel.

Animals↗

PubMed1987

SUN 1165: a new antiarrhythmic Na current blocker in ventricular myocytes of guinea-pig.

1. We compared the effect of a new antiarrhythmic compound, SUN 1165, on Na and Ca channels in papillary muscles and enzymatically dispersed single ventricular cells of guinea-pig. Action potential and contractile force in papillary muscle were measured by the conventional microelectrode technique and a strain gauge. The membrane currents were measured in internally perfused and voltage clamped cells by a single suction pipette technique. 2. In papillary muscles, SUN 1165 depressed the maximum rate of rise of action potential (Vmax) in a concentration dependent manner (IC30 = 1.7 X 10(-5) M) more markedly (about six times) than the contractile force. 3. In single ventricular cells, the Na current (INa) was reduced by the drug in a concentration dependent manner (IC30 = 9.1 X 10(-6) M). 4. It showed frequency-dependent block and the steady-state inactivation curve was shifted to more negative potentials. 5. The recovery of INa from inactivation was prolonged by SUN 1165. 6. The Ca current (ICa) was also blocked by the drug in a concentration dependent manner but much less than INa (IC30 = 5.5 X 10(-5) M). 7. These results suggested that SUN 1165 causes a selective inhibition of Na channels in guinea-pig ventricular cells at the antiarrhythmic concentrations.

Action Potentials↗