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)     

Angiotensin evokes in polyploid rat glioma cells hyperpolarization-depolarization responses and cross-desensitization with bradykinin

Angiotensins I, II and III induced a hyperpolarizing response of up to 1 min duration followed by a depolarizing response of up to 4 min when applied by pressure pulses or iontophoresis to polyploid rat glioma cells C6-4-2. The hyperpolarization (depolarization) was associated with a 50% decrease (no measurable change) in membrane resistance. The reversal potential (ca. −90 mV) of the hyperpolarization most likely points to an increased K⁺ conductance. Cells desensitized to angiotensins on application of high doses of either angiotensins or bradykinin.     

Local stimulation induced GABAergic response in rat striatal slice preparations: Intracellular recordings on QX-314 injected neurons

Gamma-aminobutyric acid (GABA)ergic responses evoked by electrical stimulation in the neostriatal slice preparation were studied in neurons injected intracellularly with Na-conductance blocker QX-314. Local stimulation elicited depolarizing postsynaptic potentials (DPSPs) in the QX-314-injected neurons when the membrane potential was morenegative than −60 mV. When DPSPs were minimized by depolirizing current injection in the QX-314-injected neuron, hyperpolarization was clearly observed following local stimulation. The maximum duration of the hyperpolarizing response to strong local stimulation was about 130 ms. The hyperpolarizing response was blocked by the addition of bicuculine or picrotoxin to the Ringer solution. Intracellular Cl^- injections produced changes in the pattern of the local stimulations-induced responses; the initial depolarizing response was followed by a relatively large amplitude long duration depolarization. The polarity of the long duration of depolarizing response could not be reversed by depolarizing currents which were normally sufficient to reverse the polarity of DPSPs in the neurons without Cl^- injection. The application of pentobarbital enhanced the amplitude and the duration of the hyperpolarizing responses. The revealed potential of the pentobarbital-enhanced response was estimated to be −60 mV. On the basis of their reversal potential, sensitivity to injected Cl^-, sensitivity to GABA blockers picrotoxin and bicuculine, and the effect of pentobarbital, these hyperpolarizing responses are shown to be GABAergic Cl^- mediated inhibitory postsynaptic potentials (IPSPs).     

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.     

Slow inhibitory postsynaptic potentials and hyperpolarization evoked by noradrenaline in the neurones of mammalian sympathetic ganglion

Slow IPSPs evoked in the neurones of rabbit isolated superior cervical ganglion by repetitive orthodromic stimulation, and a response evoked in the neurones of this ganglion by perfusion of noradrenaline, were studied using intracellular microelectrodes. Slow IPSPs were observed in 36% of neurones studied, and when investigated after treatment with D-tubocurarine and neostigmine, had a mean amplitude of 4.4 +/- 0.2 mV (mean +/- S.E.) and duration of 5 sec to 1.5 min. Two types of slow IPSPs occurring in different neurones were found. The slow IPSP of the first type was followed by a decrease in cell input resistance, was increased by depolarization and decreased by hyperpolarization of the membrane, with the reversal potential, if estimated by extrapolation method, equal to -77.8 +/- 3.3 mV. The slow IPSP of the second type was not followed by any change in cell input resistance, was increased by hyperpolarization and decreased by depolarization. The slow IPSP of the second type was reversibly blocked by phentolamine (1.4 X 10(-4) M). Noradrenaline (1 X 10(-4) M) evoked hyperpolarization or hyperpolarization followed by depolarization in 55% of the neurones studied. Hyperpolarization evoked by noradrenaline had a mean amplitude to 5.0 +/- 0.2 mV, was not followed by any change in cell input resistance, was reversibly blocked by phentolamine (1.4 X 10(-4) M), and was decreased by both depolarization and hyperpolarization of the cell membrane. It has been concluded that there are two groups of neurones in superior cervical ganglia, different with respect to the ionic mechanisms underlying the slow IPSP. In the first group of neurones the slow IPSP is probably due to an increase in potassium permeability of the membrane. The ionic mechanisms underlying the slow IPSP in the second group of neurones of noradrenaline-induced hyperpolarization remain unclear.     

On the mechanism of action of GABA in pelvic vesical ganglia: Biphasic responses evoked by two opposing actions on membrane conductance

Intracellular recording techniques were used to study the response of cat vesical pelvic ganglion neurones loaded with permeable anions to the application of GABA in vitro. In 106/127 neurones GABA evoked a biphasic response, the initial phase of which was depolarizing and associated with a conductance increase; the latter phase was hyperpolarizing and associated with a conductance decrease. The GABA evoked hyperpolarization and conductance decrease were related and behaved as though generated by closure of ion channels open in the resting membrane. The hyperpolarization had a strong inhibitory action on both spontaneous activity, and excitation evoked by depolarizing current injection and pre-synaptic nerve stimulation. Ion substitution experiments suggest that the conductance decrease is primarily to chloride ions, although other ionic species may contribute. Short iontophoretic applications of GABA-evoked monophasic depolarizing excitatory responses, even during the hyperpolarizing response evoked by perfusion of GABA, suggesting no cross-desensitization between the mechanisms generating the initial and late phases of the biphasic response.     

Calcitonin gene-related peptide evokes distinct types of excitatory response in guinea pig coeliac ganglion cells

Pressure application of calcitonin gene-related peptide (CGRP) evoked in a population of guinea pig coeliac neurons 3 types of response: a fast, a slow and a biphasic depolarization. The responses were not appreciably affected in low Ca/high Mg or tetrodotoxin-containing Krebs solution. The fast depolarization was associated with a fall in membrane resistance; it was made larger on hyperpolarization and the estimated reversal potential was -24 mV. The fast response was reversibly blocked in a Na-free medium as well as by relatively high concentrations of d-tubocurarine (50-100 microM) but not by hexamethonium. The slow, CGRP-induced depolarization resistant to nicotinic and muscarinic antagonists, was associated with either a small increase or decrease of input resistance. Membrane hyperpolarization increased the slow response in the majority of coeliac neurons, with an estimated reversal potential of -44 mV. The biphasic depolarization displayed electrophysiological and pharmacological characteristics resembling the fast and slow responses. These results raise the possibility that CGRP acting via two distinct types of receptor elicits, respectively, a fast, Na-dependent excitatory response and a slow response, the mechanism of which remains to be established.     

Ionic mechanism of a hyperpolarizing glutamate effect on two identified neurons in the buccal ganglion of Aplysia

The ionic mechanism of a membrane effect of L-glutamate on two identified neurons in the buccal ganglion of Aplysia kurodai was investigated with conventional microelectrode techniques and glutamate iontophoresis. Bath-applied and iontophoresed glutamate hyperpolarized the membrane and increased the membrane conductance. The hyperpolarizing glutamate response decreased in amplitude and finally reversed its polarity by conditioning hyperpolarization. The reversal potential of the hyperpolarizing glutamate response was close to the ECl (-60 mV). The reversal potential changed by 22.4 mV when the external chloride concentration was altered by a factor of 5. The relationship between the iontophoretically applied current and the membrane conductance changes was suggestive of two glutamate molecules reacting with a single receptor site. The hyperpolarizing glutamate response was essentially unaffected by 2-amino-4-phosphonobutyric acid (2-APB), L-proline, and quinuclidinyl benzilate (QNB). It was concluded that the hyperpolarizing glutamate response was generated by an activation of Cl- conductance.     

Intra-axonal recordings in rat dorsal column axons: membrane hyperpolarization and decreased excitability precede the primary afferent depolarization

Intra-axonal recordings were obtained in the dorsal columns of the rat lumbosacral spinal cord. Dorsal root or dorsal column stimulation at levels subthreshold for the impaled axon elicited a prolonged depolarization corresponding to the primary afferent depolarization (PAD). The depolarization was preceded by a brief hyperpolarizing potential during which excitability was decreased. The hyperpolarization corresponds temporally to the extracellularly recorded DRP IV component of the dorsal root potential described by Lloyd and McIntyre, and may represent the intracellular correlate of this potential. Possible mechanisms for this hyperpolarization include electrical interactions between neuronal elements, a biphasic GABA response, or attenuation of background afferent axonal depolarization.     

Recruitment of inhibition by enhanced activation of synaptic NMDA responses in the rat cerebral cortex

Intracellular recordings of layer V neurons from rat neocortical slices were obtained to examine the effects of reducing extracellular magnesium on inhibition. Magnesium-free solutions induced interictal and ictal-like events in cortical neurons. Changes in synaptic events underlying epileptogenesis were studied when extracellular calcium was raised (from 2 to 3–7 mM) since this delayed seizure activity. With increasing time of exposure of cells to magnesium-free solutions, there was a significant increase in the size and duration of both the depolarizing and slow synaptic hyperpolarizing responses, but the fast synaptic hyperpolarizationsignificantly declined in amplitude. When cells were recorded with cesium acetate-filled microelectrodes slow hyperpolarizing responses were blocked, but depolarization of cells to 0 mV allowed an isolated fast hyperpolarizing response to be recorded following synaptic stimulation. The amplitude of this response was unchanged after exposure to magnesium-free solutions. Synaptic responses of cells initially bathed in an (NMDA) antagonist (CPP) were unchanged by subsequent exposure to magnesium-free solutions. CPP exposure by itself caused in depolarization duration, increase in fast hyperpolarizing amplitude, and decrease in slow hyperpolarization amplitude and duration. When the fast hyperpolarization was viewed in isolation (cesium recording electrodes) at 0 mV, the amplitude of this event was unchanged by exposure to CPP. Given these results stimulus-response characteristics of neocortical neurons were reassessed under control conditions. With higher intensity stimuli larger depolarizing and slow hyperpolarizing responses were evoked, but the fast hyperpolarization showed a decremental response. These effects were reversed when CPP was added. When NMDA activity was enhanced by exposure to magnesium-free solutions or electrical stimulation, the amplitude of excitatory events and slow hyperpolarizations increased, but fast inhibitory responses showed limited capacity for incremental recruitment. This suggests fast inhibition is saturated (maximal) at submaximal levels of excitation, and can be overcome by increasing levels of excitation. Such a process is active under physiological conditions, altering the efficacy of inhibition.     

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.     

In vitro electrophysiology of rat subicular bursting neurons

Intracellular recordings were used to study the electrophysiological properties of rat subicular neurons in a brain slice preparation in vitro. Cells were classified as bursting neurons (n = 102) based on the firing pattern induced by depolarizing current pulses. The bursting response recorded at resting membrane potential (−66.1 ± 6.2 mV, mean ± SD n = 94) was made up of a cluster of fast action potentials riding on a slow depolarization and was followed by an afterhyperpolarization. Tonic firing occurred at a membrane potential of approximately −55 mV. A burst also occurred upon termination of a hyperpolarizing current pulse. Tetrodotoxin (TTX, 1 μM) blocked the burst and decreased or abolished the underlying slow depolarization. These effects were not induced by the concomitant application of the Ca²⁺ channel blockers Co²⁺ (2 mM) and Cd²⁺ (1 mM). Subicular bursting neurons displayed voltage- and time-dependent inward rectifications of the membrane during depolarizing and hyperpolarizing current pulses. The inward rectification in the depolarizing direction was abolished by TTX, while that in the hyperpolarizing direction was blocked by extracellular Cs⁺ (3 mM), but not modified by Ba²⁺ (0.5–1 mM), TTX, or Co²⁺ and Cd²⁺. Tetraethylammonium (10 mM)-sensitive, outward rectification became apparent in the presence of TTX. These results suggest that neurons in the rat subiculum can display voltage-dependent bursts of action potentials as well as membrane rectification in the depolarizing and hyperpolarizing directions. These results also indicate that activation of a voltage-gated Na⁺ conductance may be instrumental in the initiation of bursting activity. Hippocampus 7:48–57, 1997. © 1997 Wiley-Liss, Inc.     

Excitatory and inhibitory effects of epinephrine on neonatal rat sympathetic preganglionic neurons in vitro

Current and voltage recordings were made from antidromically identified sympathetic preganglionic neurons (SPNs) in transverse thoracolumbar spinal cord slices removed from neonatal rats. When applied by either pressure ejection or superfusion, epinephrine (Epi) caused a slow depolarization or an inward current in 62 SPNs (42%) and a slow hyperpolarization or an outward current in 21 SPNs (14%). The responses persisted in low calcium- or tetrodotoxin-containing media. The Epi-induced depolarization or inward current was associated with increased membrane resistance; it was reduced by membrane hyperpolarization and nullified at a membrane potential of about -100 mV; a clear reversal however was not observed at more negative potential levels. In a number of SPNs the Epi-induced depolarization was accompanied by small inhibitory postsynaptic potentials. The latter were eliminated by a low calcium solution and by the glycine antagonist strychnine, suggesting that they were caused by glycine or a glycine-like substance released from interneurons subsequent to activation by Epi. The Epi-induced hyperpolarization or outward current was associated with decreased membrane resistance, and nullified around -100 mV. The alpha-adrenergic antagonist, dihydroergotamine, and alpha 1-antagonist, prazosin, reversibly blocked the excitatory, whereas the alpha 2-antagonist, yohimbine, abolished the inhibitory response, respectively. It is concluded that Epi acting on alpha 1- and alpha 2-adrenergic receptors depolarizes and hyperpolarizes the rat SPNs by decreasing or increasing membrane conductances to potassium ions.     

Analysis of K+ accumulation reveals privileged extracellular region in the vicinity of glial cells in situ

Astrocytes and oligodendrocytes in rat and mouse spinal cord slices, characterized by passive membrane currents during de- and hyperpolarizing stimulation pulses, express a high resting K+ conductance. In contrast to the case for astrocytes, a depolarizing prepulse in oligodendrocytes produces a significant shift of reversal potential (Vrev) to positive values, arising from the larger accumulation of K+ in the vicinity of the oligodendrocyte membrane. As a result, oligodendrocytes express large tail currents (Itail) after a depolarizing prepulse due to the shift of K+ into the cell. In the present study, we used a mathematical model to calculate the volume of the extracellular space (ECS) in the vicinity of astrocytes and oligodendrocytes (ESVv), defined as the volume available for K+ accumulation during membrane depolarization. A mathematical analysis of membrane currents revealed no differences between glial cells from mouse (n = 59) or rat (n = 60) spinal cord slices. We found that the Vrev of a cell after a depolarizing pulse increases with increasing Itail, expressed as the ratio of the integral inward current (Qin) after the depolarizing pulse to the total integral outward current (Qout) during the pulse. In astrocytes with small Itail and Vrev ranging from -50 to -70 mV, the Qin was only 3-19% of Qout, whereas, in oligodendrocytes with large Itail and Vrev between -20 and 0 mV, Qin/Qout was 30-75%. On the other hand, ESVv decreased with increasing values of Vrev. In astrocytes, ESVv ranged from 2 to 50 microm3, and, in oligodendrocytes, it ranged from 0.1 to 2.0 microm3. Cell swelling evoked by the application of hypotonic solution shifted Vrev to more positive values by 17.2 +/- 1.8 mV and was accompanied by a decrease in ESVv of 3.6 +/- 1.3 microm3. Our mathematical analysis reveals a 10-100 times smaller region of the extracellular space available for K+ accumulation during cell depolarization in the vicinity of oligodendrocytes than in the vicinity of astrocytes. The presence of such privileged regions around cells in the CNS may affect the accumulation and diffusion of other neuroactive substances and alter communication between cells in the CNS.     

Cholinergic synaptic potentials in the supragranular layers of auditory cortex

Receptive-field plasticity within the auditory neocortex is associated with learning, memory, and acetylcholine (ACh). However, the interplay of elements involved in changing receptive-fields remains unclear. Herein, we describe a depolarizing and a hyperpolarizing potential elicited by repetitive stimulation (20-100 Hz, 0.5-2 sec) and dependent on ACh, which may be involved in modifying receptive-fields. These potentials were recorded, using whole cell techniques, in layer II/III pyramidal cells in the rat auditory cortex in vitro. Stimulation at low stimulus intensities can give rise to a hyperpolarizing response and stimulation at higher stimulus intensities can elicit a depolarizing response. The depolarizing response had a reversal potential of -35 mV, and was reduced by the combination of AMPA/kainate and NMDA glutamate receptor antagonists (AMPA/kainate: CNQX, DNQX, and GYKI 52466; NMDA: APV, MK-801) and by the muscarinic ACh receptor antagonist atropine. The hyperpolarizing response had a reversal potential of -73 mV and could be reduced by atropine, GABA(A) receptor antagonists (bicuculline and a Cl(-) channel blocker picrotoxin), and to a small extent a GABA(B) receptor antagonist (saclofen). This suggests that the hyperpolarizing response is likely to be mediated by ACh acting on GABAergic interneurons. Extracellular recordings, also made from layer II/III of cortical slices, yielded a negative-going potential which was reduced by ionotropic glutamate receptor antagonists (same as above) and by the ACh receptor antagonists atropine and scopolamine, suggesting that this potential was the extracellular representation of the depolarizing response.     

Synaptic response of bulbar respiratory neurons to hypercapnic stimulation in peripherally chemodenervated cats.

Effects of hypercapnia on the membrane potential and synaptic activity of bulbar respiratory neurons were studied in decerebrate, vagotomized, glomectomized and artificially ventilated cats. Coaxial multibarrelled electrodes were used for intracellular recording and extracellular iontophoresis of drugs. Hypoventilation with oxygen-enriched air (hyperoxic hypercapnia) produced an increase of depolarization together with an increase of spiking during the active phase and an increase of hyperpolarization during the inactive phase of each respiratory cycle in the inspiratory, postinspiratory and expiratory neurons of the ventral respiratory group. Both depolarizing and hyperpolarizing effects were associated with a decrease in input resistance. Intracellular injection of Cl- reversed the polarity of the hyperpolarizing synaptic wave to depolarization during the inactive phase, and hypercapnia increased the depolarization at that phase. Iontophoresis of tetrodotoxin eliminated the CO2-induced changes in membrane potential and input resistance. In 20 out of 58 neurons examined, iontophoretically applied atropine partly or totally suppressed the depolarizing response to hypercapnia. For these neurons, iontophoresed acetylcholine produced a sustained depolarization that was antagonized by atropine, but not by hexamethonium. The present study shows that both depolarizing and hyperpolarizing responses of medullary respiratory neurons to hyperoxic hypercapnia are synaptically mediated. A muscarinic mechanism is involved in part of the respiratory neuronal excitation evoked by hypercapnia.     

GABAA receptor stimulation blocks NMDA-induced bursting of dopaminergic neurons in vitro by decreasing input resistance.

The effects of the GABAA agonist, isoguvacine, on NMDA-induced burst firing of substantia nigra dopaminergic neurons were studied with intracellular and whole cell recordings in vitro. NMDA application caused the neurons to fire in rhythmic bursts. Although the NMDA-induced bursty firing pattern was insensitive to hyperpolarization by current injection, it was reversibly abolished by the selective GABAA agonist, isoguvacine. The block of the rhythmic burst pattern by isoguvacine application occurred regardless of whether the chloride reversal potential was hyperpolarizing (ECl-=-70 mV) or depolarizing (ECl-=-40 mV). In either case, the input resistance of the dopaminergic neurons was dramatically decreased by application of isoguvacine. It is concluded that GABAA receptor activation by isoguvacine disrupts NMDA receptor-mediated burst firing by increasing the input conductance and thereby shunting the effects of NMDA acting at a distally located generator of rhythmic burst firing.     

The dendritic response to GABA in CA1 of the hippocampal slice

Application of GABA in the dendritic region of pyramidal cells elicits a depolarization which, in fact, is the sum of a hyperpolarizing and a depolarizing process. At the reversal potential of the depolarizing response (-42 mV) the GABA-induced current fluctuations do not have a minimum. Consequently, a conductance change to more than one ion is involved. Cl- is in part responsible, Ca2+ is not because Mn2+ and Mg2+ do not change the response. Whether Na+ is involved is uncertain. Substitution with choline had no effect but choline may permeate through the membrane during the depolarizing response. Nipecotic acid inhibits a Na+-GABA uptake mechanism but does not change the dendritic response.     

Cholinergic synapses and the organization of contrast detection in the crayfish optic lobe

The actions of acetylcholine (ACh) were examined on 4 classes of multicolumnar interneurons whose dendrites lie in close proximity to the putative cholinergic transmedullary neurons described in the companion report. ACh-elicited responses in each cell type resemble visually elicited synaptic events and persist following synaptic blockade with 20 mM CoCl2. Tangential cells exhibit a hyperpolarizing response to ACh that resembles the visual response in reversal potential and dependence on extracellular chloride. The visual response is potentiated by the anticholinesterase, neostigmine (0.1 mM). Visual and carbachol-elicited responses are blocked by nicotinic ganglionic antagonists (e.g., 10(-6) M pempidine) that are 10-100 times more potent than D-tubocurarine. Medullary amacrine cells exhibit depolarizing responses to ACh (10(-6) M) and light with similar reversal potentials. The visual response is potentiated by neostigmine. Dimming fibers respond to light and ACh with a hyperpolarization that inhibits the maintained discharge. The sustaining fiber response to ACh reflects both direct responses and indirectly elicited synaptic actions. The direct action is a hyperpolarization possibly related to the visual "off-response." It is associated with an increased conductance and a reversal potential negative to the dark potential. The off-response is abolished by curare and pempidine and potentiated by neostigmine. ACh appears to orchestrate several aspects of the dual-channel contrast detection system of the optic lobe. The actions of ACh on tangential cells, amacrine cells, and dimming fibers are all consistent with the effects of a spatially localized increment in light intensity and a corresponding local release of ACh in the retinotopic columnar array.     

Changes of the membrane potential in striatal synaptoneurosome, synaptosome and membrane sac preparations induced by glutamate, kainate and aspartate as measured with a cyanine dye DiS-C2-(5)

The effects of glutamate, kainate and aspartate on the membrane potential of striatal synaptoneurosome, synaptosome and membrane sac preparations were studied by using a potential sensitive cyanine dye DiS-C2-(5). Excitatory amino acids glutamate and aspartate had a depolarizing effect on synaptoneurosomes. 7.9 microM glutamate and 2.8 microM aspartate produced a half-maximal response. Depolarizations induced by glutamate and aspartate were dependent on the concentration of extracellular sodium ions, a maximal response occurred at around 40 mM of external Na+. Kainate induced a dual effect on synaptoneurosomes. In a standard Na+-based medium a hyperpolarization, likely due to inhibition of a presynaptic sodium-dependent glutamate uptake, predominated over a postsynaptic kainate receptor-mediated depolarization that was observed when electrogenic glutamate uptake was inhibited. This interpretation was supported by results obtained with synaptosome and membrane sac preparations. In a standard Na+-based medium kainate had a hyperpolarizing effect on synaptosomes while in the membrane sac preparation kainate induced a depolarization.