In vitro studies of the role of gamma-aminobutyric acid in inhibition in the lateral septum of the rat

Focal stimulation, stimulation of the fimbria, and stimulation of the medial septal area result in an inhibitory postsynaptic potential (IPSP) in lateral septal neurons. Increased stimulus intensity results in the appearance of a late hyperpolarizing potential (LHP). Treatment of the slice with bicuculline methiodide or picrotoxin results in blockade of the IPSP. When present, LHPs are enhanced in the presence of bicuculline or picrotoxin. Spontaneous and evoked IPSPs reverse near -70 mV, and LHPs reverse near -90 mV. Iontophoretic application of gamma-amino-butyric acid (GABA) results in hyperpolarizing, depolarizing, or biphasic potentials. Treatment with bicuculline or picrotoxin results in depression of biphasic GABA responses that appears selective for the depolarizing portion of the potential. At high concentrations of bicuculline, a portion of the hyperpolarizing GABA potential persists. The reversal potential of the depolarizing GABA potential is near -30 mV, and the reversal potential of monophasic hyperpolarizing GABA potential is near -70 mV. The bicuculline-resistant hyperpolarizing GABA response has a reversal potential near -90 mV. GABA activates three separate conductances on septal neurons, which are similar to those reported on hippocampal neurons. The resistance of the hyperpolarizing GABA potential to bicuculline appears to be due to the presence of a GABA-activated potassium conductance, which is similar to that activated by baclofen.     

Whole-cell patch study of GABAergic inhibition in CA1 neurons of immature rat hippocampal slices

Inhibitory processes mediated by gamma-aminobutyric acid (GABA) were studied in immature rat hippocampal slices using the whole-cell patch clamp technique. Orthodromically evoked hyperpolarizing inhibitory postsynaptic potentials (IPSPs) were observed in CA1 neurons of postnatal 2-5 (P2-5) and 7-13 (P7-13) day old rats under conditions of low internal [Cl-]. In the whole-cell voltage-clamp mode, applications of GABA evoked outwards currents which reversed at -55 mV and -62 mV in P2-5 and P7-13 CA1 neurons, respectively, with comparable reversal potential for the IPSPs for each age. An increase in internal [Cl-] caused a depolarizing shift of the GABA reversal potential which followed the Nernst equation. In both groups of neurons, the IPSPs and GABA currents were blocked with the bath applications of bicuculline (10 microM) and picrotoxin (100 microM). We conclude that the GABAA-mediated inhibitory synaptic process exists in P2-5 CA1 neurons and hypothesize that the absence of such IPSPs noted in previous studies of immature CA1 neurons was likely due to higher internal [Cl-] in the more immature neurons.     

Synaptic inhibition in the isolated respiratory network of neonatal rats

Gramicidin-perforated patch-clamp recording revealed phasic Cl^–-mediated hyperpolarizations in respiratory neurons of the brainstem–spinal cord preparation from newborn rats. The in vitro respiratory rhythm persisted after block of γ-aminobutyric acid (GABA), i.e. GABA_A, receptor-mediated inhibitory postsynaptic potentials (IPSPs) with bicuculline and/or glycinergic IPSPs with strychnine. In one class of expiratory neurons, bicuculline unmasked inspiration-related excitatory postsynaptic potentials (EPSPs), leading to spike discharge. Bicuculline also blocked hyperpolarizations and respiratory arrest due to bath-applied muscimol, whereas strychnine antagonized similar responses to glycine. The reversal potential of respiration-related IPSPs and responses to GABA, muscimol or glycine was not affected by CO₂/HCO₃^–-free solutions, but shifted from about ?65 mV to values more positive than ?20 mV upon dialysis of the cells with 144 instead of 4 mm Cl^–. Impairment of GABA uptake with nipecotic acid or glycine uptake with sarcosine evoked a bicuculline- or strychnine-sensitive decrease of respiratory frequency which could lead to respiratory arrest. Also, the GABA_B receptor agonist baclofen led to reversible suppression of respiratory rhythm. This in vitro apnoea was accompanied by a K⁺ channel-mediated hyperpolarization (reversal potential ?88 mV) of tonic cells, whereas membrane potential of neighbouring respiratory neurons remained almost unaffected. Both baclofen-induced hyperpolarization and respiratory depression were antagonised by 2-OH-saclofen, which did not affect respiration-related IPSPs per se. The results show that synaptic inhibition is not essential for rhythmogenesis in the isolated neonatal respiratory network, although (endogenous) GABA and glycine have a strong modulatory action. Hyperpolarizing IPSPs mediated by GABA_A and glycine receptors provide a characteristic pattern of membrane potential oscillations in respiratory neurons, whereas GABA_B receptors rather appear to be a feature of non-respiratory neurons, possibly providing excitatory drive to the network.     

Effects of glycine on neurons in the rat substantia nigra zona compacta: in vitro electrophysiological study

The effects of bath-applied glycine to substantia nigra zona compacta neurons of rat were investigated by intracellular recording techniques in vitro. Superfusion of glycine (1 mM) in the medium hyperpolarized 53% of the neurons recorded with KCl electrodes, whereas 32% of the cells were depolarized. The remaining 15% of neurons was hyperpolarized and then depolarized by the amino acid. In spite of these membrane changes, the action potential firing was depressed. Both hyperpolarization and depolarization were correlated to an outward and an inward current, respectively, when recording in single-electrode voltage-clamp mode. In response to bath application of glycine, the neurons showed a concentration-dependent conductance increase. Micromolar concentrations of glycine (100-300 microM) in the superfusion medium produced a membrane hyperpolarization (outward current) in most of the tested cells, whereas millimolar concentration of amino acid could cause depolarization (inward current) in the same neurons. When the recording electrodes contained K acetate, only hyperpolarizations (outward current) were produced. The potential and current changes and the increase in membrane conductance produced by glycine showed little desensitization when neurons were recorded with K acetate electrodes. The mean reversal potential for the membrane hyperpolarization was -80 mV with intracellular electrodes containing KCl and -84 mV with electrodes containing K acetate. The mean null potential for the depolarizing effect was -46 mV. The reversal potential for the glycinergic responses was shifted to less negative values upon lowering the extracellular concentration of chloride or increasing the extracellular concentration of potassium. Strychnine (1-10 microM) reversibly antagonized both the conductance increase and the membrane changes produced by glycine. Bath application of bicuculline (100 microM) and picrotoxin (200 microM) did not affect glycine responses, while depressing the actions of GABA and muscimol. It is concluded the glycine exerts an inhibition on substantia nigra zona compacta neurons by acting on strychnine-sensitive receptors. The membrane effects produced by glycine result from the activation of a chloride current. In addition, the simultaneous involvement of potassium ions may contribute to the overall effects of glycine.     

GABA mediated excitation in immature rat CA3 hippocampal neurons

Intracellular recordings from rat hippocampal neurons in vitro during the first postnatal week revealed the presence of spontaneous giant depolarizing potentials (GDPs). These were generated by the synchronous discharge of a population of neurons. GDPs reversed polarity at -27 and -51 mV when recorded with KCl or K-methylsulphate filled electrodes, respectively. GDPs were blocked by the GABAA receptor antagonist bicuculline (10 microM). Iontophoretic or bath applications of GABA (10-300 microM) in the presence of tetrodotoxin (1 microM), induced a membrane depolarization or in voltage clamp experiments an inward current which reversed polarity at the same potential as GDPs. The response to GABA was blocked in a non-competitive manner by bicuculline (10 microM) and did not desensitize. GABA mediated GDPs were presynaptically modulated by N-methyl-D-aspartate (NMDA) and non-NMDA receptors. Their frequency was reduced or blocked by NMDA receptor antagonists and by the rather specific non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). The frequency of GDPs was enhanced by glycine and D-serine (10-30 microM) in a strychnine insensitive manner. This effect was blocked by AP-5, suggesting that it was mediated by the allosteric modulatory site of the NMDA receptor. These observations suggest that most of the 'excitatory' drive in immature neurons is mediated by GABA acting on GABAA receptors; furthermore excitatory amino acids modulate the release of GABA by a presynaptic action on GABAergic interneurons.     

Early development of glycine- and GABA-mediated synapses in rat spinal cord.

Motoneuron responses to the inhibitory amino acids glycine and GABA, and the contribution of inhibitory synapses to developing sensorimotor synapses were studied in rat spinal cords during the last week in utero. In differentiating motoneurons, glycine and GABA induced Cl(-)-dependent membrane depolarizations and large decreases in membrane resistance. These responses gradually decreased during embryonic development, and at birth they were significantly smaller than in embryos. In motoneurons of embryos and neonates, dorsal root stimulation produced only depolarizing potentials, some of which reversed at -50 mV membrane potential. Reduction of extracellular Cl- concentrations increased the amplitude of these potentials, suggesting that they are generated by Cl- current. Contribution of Cl(-)-dependent potentials to compound dorsal root-evoked potentials was studied by determining the effects of glycine and GABA antagonists on them. In motoneurons of embryos at days 16-17 of gestation (D16-D17), strychnine or bicuculline blocked dorsal root-evoked potentials. This suppression was neither the result of a decrease in neuronal excitability nor the inhibition of glutamate receptors. Strychnine-evoked depression was not blocked by atropine, indicating that it was not due to disinhibition of muscarinic synapses. By D19, strychnine and bicuculline significantly increased dorsal root-evoked potentials rather than blocking them. This reversed function did not result from an increase in neuronal excitability or changes in the specificity of strychnine and bicuculline antagonism. The number of glycine- and GABA-immunoreactive cells increased 20% between D17 and D19. The number of immunoreactive cells and fibers significantly increased in the motor nuclei and dorsal horn laminae. These morphological changes may contribute to establishment of new synaptic contacts on motoneurons, thus changing the actions of strychnine and bicuculline on dorsal root-evoked potentials.     

gamma-Aminobutyric acid (GABA) causes consistent depolarization of neurons in the guinea pig supraoptic nucleus due to an absence of GABAB recognition sites

The action of gamma-aminobutyric acid (GABA) in the supraoptic nucleus was investigated using guinea pig brain slices. GABA produced a membrane depolarization accompanied by a decrease in the input resistance. The action of GABA was concentration-dependent throughout a wide range of concentrations (10(-7)-10(-3) M). In none of the cells examined, a membrane hyperpolarization was observed. The reversal potential for the depolarization induced by GABA was about 25 mV positive to the resting membrane potential. The amplitude of the GABA-induced depolarization was increased to 1.5 X the control by reducing the external Cl- from 134.2 mM to 10.2 mM. The action of GABA was readily antagonized by relatively low concentrations of bicuculline (10(-5) M). The action of GABA in the hippocampus or in the anterior hypothalamus was markedly different from that in the supraoptic nucleus, i.e. GABA produced both depolarizing and hyperpolarizing responses in the hippocampus and consistently a hyperpolarization in the anterior hypothalamus. The depolarizing but not the hyperpolarizing response in the hippocampus was selectively blocked by picrotoxin (2 X 10(-5) M) or by bicuculline (10(-5) M). The depolarizing component was dependent on the external Cl- concentration and had a reversal potential similar to that of the depolarization induced by GABA in the supraoptic nucleus. The hyperpolarizing component was resistant to bicuculline and had a reversal potential about 30 mV negative to the resting membrane potential.(ABSTRACT TRUNCATED AT 250 WORDS)     

Currents evoked by GABA and glycine in acutely dissociated neurons from the rat medial preoptic nucleus

The responses of acutely dissociated medial preoptic neurons to application of GABA and glycine were studied using the perforated-patch whole-cell recording technique under voltage-clamp conditions. GABA, at a concentration of 1 mM, evoked outward currents in all cells (n=33) when studied at potentials positive to −80 mV. The I–V relation was roughly linear. The currents evoked by GABA were partially blocked by 25–75 μM picrotoxin and were also partially or completely blocked by 100–200 μM bicuculline. Glycine, at a concentration of 1 mM, did also evoke outward currents in all cells (n=12) when studied at potentials positive to −75 mV. The I–V relation was roughly linear. The currents evoked by glycine were largely blocked by 1 μM strychnine. In conclusion, the present work demonstrates that neurons from the medial preoptic nucleus of rat directly respond to the inhibitory transmitters GABA and glycine with currents that can be attributed to GABA_A receptors and glycine receptors respectively.     

Beta-alanine acts on glycine receptors in the rat sacral dorsal commissural neurons

Whole-cell current responses to bath application of beta-alanine (beta-ALA) were investigated in neurons acutely dissociated from the rat sacra! dorsal commissural nucleus (SDCN) using the nystatin perforated patch recording configuration under voltage-clamp conditions. beta-ALA activated inward currents in a concentration-dependent manner over the range of 10-3000 microM with an EC50 of 80.8 microM. The reversal potential of beta-ALA-activated currents (I beta-ALA) was close to the Cl- equilibrium potential. Strychnine and the chloride channel blocker picrotoxin suppressed the I beta-ALA in a concentration-dependent manner with the IC50 values of 0.19 microM and 343.6 microM, respectively. At the concentration of 3 microM, strychnine was sufficient to completely block 100 microM beta-ALA response, whereas it did not show a suppression of GABA response. In contrast, bicuculline, a potent GABAA receptor antagonist, at concentrations up to 10 microM, a dose that could block the GABA response completely, had little or no effect on I beta-ALA. Furthermore, the I beta-ALA was not affected by a preceding GABA response, but rather cross-desensitized with that evoked by glycine. The results indicate that beta-ALA activates the strychnine-sensitive glycine receptors in the SDCN neurons, and suggest that beta-ALA may act as a functional neurotransmitter in the mammalian SDCN.     

Sensitivity of identified medial hypothalamic neurons to GABA, glycine and related amino acids; influence of bicuculline, picrotoxin and strychnine on synaptic inhibition

Medial hypothalamic neurons in pentobarbital anesthetized rats were identified by location and response to electrical stimulation of the amygdala, medial preoptic area, midbrain periaqueductal gray and median eminence. Cells were then examined for their sensitivity to microiontophoretic applications of GABA, glycine and 3 related amino acids, i.e. β-guanidinopropionic acid, δ-aminovaleric acid and β-alanine.Application of all agents decreased the spontaneous and glutamate or aspartate evoked activity of the majority of neurons in all identified categories.The majority of neurons were more sensitive to β-guanidinopropionate, δ-amminovalerate and GABA than to glycine and β-alanine.Bicuculline and picrotoxin produced a selective and reversible antagonism of depressions evoked by GABA and GABA-like amino acids whereas strychnine produced a selective and reversible antagonism of depression evoked by glycine and β-alanine.Bicuculline and picrotoxin, but not strychnine, were observed to diminish synaptic inhibition evoked by electrical stimulation of several sites when these agents were administered by microiontophoresis and by i.v. injections in convulsive doses.These observations suggest that many medial hypothalamic neurons have both GABA and glycine receptors but that GABA may have the more important role as a neurotransmitter common to afferent or recurrent inhibitory hypothalamic pathways.     

Taurine activates strychnine-sensitive glycine receptors in neurons of the rat inferior colliculus

Taurine (Tau) is one of the most abundant free amino acids in the mammalian central nervous system. Whether the neurotransmission of the central auditory system is regulated or modulated by Tau is not clear. In the present study, we investigated the electrophysiological and pharmacological properties of Tau-activated currents in acutely dissociated neurons of the rat inferior colliculus (IC) using whole cell patch clamp recordings. At a holding potential of -60 mV and under a condition of chloride equilibrium potential near 0 mV, Tau activated an inward current and its half-maximal activation concentration was equal to 0.37 mM. The measured reversal potential of Tau-activated currents was close to theoretical chloride equilibrium potential. The currents evoked by Tau at both low (1 mM) and high (10 mM) concentrations were almost completely inhibited by strychnine, a glycine receptor antagonist. The Tau-activated current, however, was not affected by bicuculline, a GABA(A) receptor antagonist. Tau at increased concentrations progressively reduced the current response to subsequent glycine application. At saturated concentrations, Tau-activated current and glycine-activated current were mutually cross-desensitized by each other. These findings indicate that Tau activates glycine receptors in neurons of the rat IC and thus may have a functional role in regulating or modulating the neurotransmission of the central auditory system in mammals.     

Fast inhibitory postsynaptic potentials and responses to inhibitory amino acids of sympathetic preganglionic neurons in the adult cat.

Intracellular recordings were obtained from sympathetic preganglionic neurons (SPNs) of the intermediolateral nucleus (IML) in slices of upper thoracic spinal cord of the anesthetized cat. A total of 44 neurons was studied. Single shock stimulation of an area of white matter dorsolateral to the IML, close to the recording electrode (< 0.5 mm), evoked fast IPSPs with rise time of 3.8 ms and 1/2 decay time of 14.7 ms (n = 12). In 17 other cells only fast EPSPs were recorded but, after suppression of the EPSPs by the excitatory amino acid receptor antagonists CNQX (20 microM) and APV (100-250 microM), fast IPSPs were unmasked. The IPSP reversed polarity at -63 mV (-67 mV in the presence of CNQX and APV). The reversal potential shifted to a less negative value when the extracellular chloride concentration was reduced. The IPSP was reversibly abolished by the GABAA receptor antagonist bicuculline in 32% of the cells, by the glycine receptor antagonist strychnine in 47% of the cells and by the combination of the two in 21% of the cells. The IPSP was abolished by TTX (0.5 microM), had constant latency and showed no failures during high frequency stimulation. The IPSP presumably resulted from the excitation of inhibitory axons and/or inhibitory neuron somata with monosynaptic connections to the SPN. Glycine and GABA (1-3 mM) produced hyperpolarization associated with decreased membrane resistance. Sixty-nine percent of cells responded to both agonists, 19% to glycine only and 12% to GABA only. The GABAB agonist baclofen (5 microM) had no effect.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibitory synaptic transmission in area postrema neurons of the rat showing robust presynaptic facilitation mediated by nicotinic ACh receptors

Inhibitory synaptic transmission and its modulation in neurons of the area postrema (AP), one of autonomic nuclei in the medulla, were studied using whole-cell patch-electrodes applied to slices from rats on postnatal days 10-24. When glycine (100 microM) or GABA (10 microM) was applied to AP neurons from a "Y tube", large outward currents that showed reversal potential of - 67 mV (approximate Cl- equilibrium potential estimated) were induced. At a holding potential of - 10 mV, application of high K+ to the AP neurons evoked massive inhibitory postsynaptic currents (IPSCs) in the neurons. Most of the evoked synaptic currents were blocked by bicuculline, while the remaining currents were sensitive to strychnine, indicating that the major inhibitory transmission in the area postrema was GABAergic. When nicotine (5-100 microM) was applied to AP neurons, robust IPSCs having GABAergic identity were evoked. Even in the presence of tetrodotoxin, nicotine could induce GABAergic IPSCs, most of which, however, disappeared in the presence of 5 mM Mg2+. Presynaptic facilitation was also induced by other nicotinic agonists, including cytisine, 1,1-dimethyl-4-phenyl-piperazinium iodide, ACh and choline. The nicotine-induced presynaptic facilitation was inhibited by mecamylamine and slightly inhibited by dihydro-beta-erythroidine or alpha-Bungarotoxin. These results indicate that nicotinic receptors are expressed at GABAergic presynaptic terminals in the area postrema and induce Ca2+ influx to trigger vesicular release. The major nicotinic receptors involved are thought to be heteromeric subtypes such as alpha3beta4 receptors, which may regulate inhibitory transmission potently responding to endogenous or exogenous nicotinic agents appeared in this area.     

Transmitters and reticulospinal neurons

Glycine and GABA strongly depress the firing of reticulospinal neurons, glycine being more effective than GABA. The effects of strychnine and bicuculline methochloride on synaptic inhibition and on the actions of these amino acids are consistent with the earlier evidence that both glycine and GABA are inhibitory transmitters in the medullary reticular formation. However, although strychnine is consistently a relatively specific antagonist of glycine, bicuculline methochloride is far less specific in distinguishing between glycine and GABA on reticulospinal neurons. Studies of synaptic inhibition in the brain stem using bicuculline methochloride alone are therefore to be interpreted with caution. Both convulsants should be tested on the actions of glycine and GABA before conclusions about transmitter identity are made. Large propertions of reticulospinal neurons appear to be unaffected by ACh, 5-HT, NA, and DA. Of the amine-sensitive reticulospinal neurons, ACh and DA usually depress firing whereas 5-HT and NA usually cause excitation. These effects, however, are generally weak (especially excitation), and it is difficult to draw firm conclusions about the possible functional significance of these substances on reticulospinal neurons.     

Amino acid receptors from insect muscle: electrophysiological characterization in Xenopus oocytes following expression by injection of mRNA

Poly(A)+ Messenger ribonucleic acid (mRNA) was extracted from leg muscles of the locust Schistocerca gregaria and injected into oocytes of Xenopus laevis. After 5-10 days incubation, receptors for L-glutamate, L-quisqualate, DL-ibotenate and gamma-aminobutyric acid (GABA) were expressed. Agonist-induced currents were dose-dependent, and, in the concentration range 1 microM to 1 mM, generally had peak values of 50 nA. The responses to all agonists, apart from GABA, exhibited desensitization which could not be reversed even by prolonged washing with Ringer. Application of 100 microM GABA to oocytes voltage clamped at -60 mV produced a smooth inward current with a reversal potential of -22 +/- 1 mV, which is consistent with the involvement of chloride ions. At 100 microM, picrotoxin reversibly abolished this current, while 100 microM bicuculline had no effect. L-Glutamate elicited a smooth current with a reversal potential of -52 +/- 3 mV. L-Quisqualate elicited an inward current at -60 mV with a reversal potential of -9 +/- 2 mV; this current occasionally had an oscillatory component. The response to ibotenate comprised a smooth inward current with a reversal potential of -21 +/- 3 mV which was probably mediated by chloride ions.     

Cerebellar macroneurons in microexplant cell culture. Postsynaptic amino acid pharmacology

Cerebellar neurons derived from 17- to 19-day-old fetal rats have been grown in a monolayer in microexplant cell culture, and intracellular recording coupled with iontophoresis of amino acid neurotransmitters has been employed to characterize their amino acid chemosensitivity. Although these cultures contain at least 3 different neuronal cell types, intracellular recordings were obtained from large neurons (diameter greater than 15 microns) with 1-5 dendritic shafts and fine dendritic arborizations and which could, on morphological grounds, be identified as Purkinje cells. All neurons with resting membrane potentials greater than 25 mV and with action potentials evoked by intracellular stimulation, responded to iontophoretically applied glutamate and GABA. There was essentially no chemosensitivity to glycine, beta-alanine or taurine. Aspartate application evoked only small responses at high iontophoretic currents. GABA reversibly increased membrane conductance and produced hyperpolarization at resting membrane potential with reversal potentials between -50 and -40 mV (5-10 mV more negative than resting membrane potential). Glutamate reversibly increased membrane conductance and produced depolarizing responses with extrapolated reversal potentials between 0 and -10 mV. Aspartate augmented glutamate responses at low iontophoretic currents which did not directly alter membrane potential or conductance. Thus Purkinje cells grown in the absence of parallel fiber and climbing fiber input develop autonomous neuropharmacologic specificity similar to that of Purkinje cells in vivo.     

Inhibitory postsynaptic actions of taurine, GABA and other amino acids on motoneurons of the isolated frog spinal cord.

The actions of glycine, GABA, alpha-alanine, beta-alanine and taurine were studied by intracellular recordings from lumbar motoneurons of the isolated spinal cord of the frog. All amino acids tested produced a reduction in the amplitude of postsynaptic potentials, a blockade of the antidromic action potential and an increase of membrane conductance. Furthermore, membrane polarizations occurred, which were always in the same direction as the IPSP. All these effects indicate a postsynaptic inhibitory action of these amino acids. When the relative strength of different amino acids was compared, taurine had the strongest inhibitory potency, followed by beta-alanine, alpha-alanine, GABA and glycine. Topically applied strychnine and picrotoxin induced different changes of post-synaptic potentials, indicating that distinct inhibitory systems might be influenced by these two convulsants. Interactions with amino acids showed that picrotoxin seletively diminished the postsymaptic actions of GABA, while strychnine reduced the effects of taurine, glycine, alpha- and beta-alanine. But differences in the susceptibility of these amino acid actions to strychnine could be detected: the action of taurine was more sensitively blocked by strychnine compared with glycine, alpha- and beta-alanine. With regard to these results the importance of taurine and GABA as transmitters of postsynaptic inhibition on motoneurons in the spinal cord of the frog is discussed.     

Effects of strychnine,bicuculline, and picrotoxin on inhibition of hypoglossal motoneurons

Postsynaptic potentials, evoked by lingual or hypoglossal nerve stimulation, were recorded from hypoglossal motoneurons of the cat with glass microelectrodes. Lingual nerve-evoked inhibitory postsynaptic potentials (LIPSPs) were recorded in 98% of hypoglossal motoneurons. Hypoglossal nerve stimulation caused a hyperpolarzing potential following the antidromic spike in all hypoglossal motoneurons tested. This potential was unaffected by depolarizing or hyperpolarizing currents, could not be evoked at a stimulus strength less than that which was threshold for the antidromic action potential, and did not change in shape or amplitude at stimulus strengths which were above threshold for antidromic invasion. This hyperpolarpolarizing potential was therefore considered to be an afterhyperpolarization. However, hypoglossal nerve-induced inhibitory postsynaptic potentials were recorded from hypoglossal units which had characteristics of interneurons, thus suggesting the presence of afferent fibers in the hypoglossal nerve. The hypoglossal nerve-induced afterhyperpolarization was not affected by strychnine, bicuculline, or picrotoxin. The LIPSP was antagonized by strychnine but unaffected by bicuculline or picrotoxin. The results suggest that inhibition of hypoglossal motoneurons via the lingual nerve is more likely to be mediated by glycine than gamma-aminobutyric acid (GABA) and is therefore similar to the strychnine-sensitive postsynaptic inhibition of spinal motoneurons.     

The effect of glycine and gamma-aminobutyric acid on the excitatory postsynaptic potentials of the spinal motor neurons in the lamprey in the presence of antagonists

Effect of bath application of the inhibitory amino acids (glycine and GABA) on motoneurons of EPSPs was studied in the normal physiological solutions and after preliminary administration of antagonists: strychnine (10(-6) mol/l), bicuculline (10(-4) mol/l) or picrotoxin (10(-4) mol/l). All these antagonists diminished the depression of monosynaptic EPSPs which were elicited by both amino acids (glycine and GABA). Data obtained in this study and previously reported ones permit concluding that motoneuron membranes in the spinal cord of lamprey possess the unit receptor channel complex sensitive to both amino acids.     

Ethanol selectively enhances the hyperpolarizing component of neocortical neuronal responses to locally applied GABA

Local application of GABA to rat cerebral cortical neurons in brain slices elicited biphasic responses mediated via GABA_A receptors. The fast component of the response, which was most apparent with somatic application of GABA, was hyperpolarizing at the normal resting membrane potential (GABA_h response). The slower component could be elicited by GABA application to nearly all regions of the cell, and was depolarizing at the resting membrane potential (GABA_d response). The reversal potential of evoked IPSCs recorded with whole-cell patch electrodes (−68 mV) was comparable to the reversal potential of the GABA_h response (−69 mV), and was significantly different from the reversal potential of the GABA_d response (−56 mV). The GABA_d response was more sensitive to enhancement by pentobarbital and more readily antagonized by both bicuculline and picrotoxin than the GABA_h response. Recording in bicarbonate-free buffer changed the reversal potential of the GABA_d response significantly, but had no effect on the GABA_h response. In contrast, superfusion with ethanol significantly enhanced the GABA_h response, while having no effect on the GABA_d component. Although a localized collapse of the Cl⁻ gradient, which has been proposed to underlie the GABA_d response, could explain the greater sensitivity of the GABA_d response to pentobarbital and the GABA_A antagonists, this could not account for the greater sensitivity of the GABA_h response to ethanol. Differences in GABA_A receptor subunit composition may result in the expression of dendritic and somatic GABA_A receptors that have different kinetics, reversal potentials, and sensitivity to pharmacological agents, including ethanol.