Hyperpolarizing Postsynaptic Potentials Research Articles

Основные нервные процессы – возбуждение и торможение как адаптивные реакции (обзор). Сообщение 1. Критический анализ эволюции теорий об основных нервных процессах и доказательство зависимости тормозного ответа нейронов от их текущего адаптационного состояния

In the work is criticized the paradigm about excitation and inhibition as the main nervous processes caused by the activity of specific excitatory and inhibitory systems. Analysis of the literature shows that the currently in neurophysiology dominant approach, justifying the existence of such systems, relies mainly on facts obtained in vitro, which are far from normal for the existence of nerve cells. Literature data are presented that indicate the dependence of the nature of the neuron response on its current adaptive (biological) state and the cytotoxicity of the neurotransmitter. In particular, in the review (part 1) it is shown that in response to the action of GABA (less toxic than neurotransmitter glutamate), brain neurons in both good (in vivo) and relatively bad (which is often in vitro) vital state will respond by hyperpolarizing postsynaptic potential (HPSP), either without excitation – the phenomenon of hyperpolarizational inhibition, or with the generation of nerve impulses, by posthyperpolarization recoil mechanism. In very poor living conditions (in vivo or in vitro), the neuronal response to GABA will always be a depolarizing postsynaptic potential (DPSP) with or without generation of action potentials (AP).

Perirhinal cortex hyperexcitability in pilocarpine‐treated epileptic rats

The perirhinal cortex (PC), which is heavily connected with several epileptogenic regions of the limbic system such as the entorhinal cortex and amygdala, is involved in the generation and spread of seizures. However, the functional alterations occurring within an epileptic PC network are unknown. Here, we analyzed this issue by using in vitro electrophysiology and immunohistochemistry in brain tissue obtained from pilocarpine-treated epileptic rats and age-matched, nonepileptic controls (NECs). Neurons recorded intracellularly from the PC deep layers in the two experimental groups had similar intrinsic and firing properties and generated spontaneous depolarizing and hyperpolarizing postsynaptic potentials with comparable duration and amplitude. However, spontaneous and stimulus-induced epileptiform discharges were seen with field potential recordings in over one-fifth of pilocarpine-treated slices but never in NEC tissue. These network events were reduced in duration by antagonizing NMDA receptors and abolished by NMDA + non-NMDA glutamatergic receptor antagonists. Pharmacologically isolated isolated inhibitory postsynaptic potentials had reversal potentials for the early GABA(A) receptor-mediated component that were significantly more depolarized in pilocarpine-treated cells. Experiments with a potassium-chloride cotransporter 2 antibody identified, in pilocarpine-treated PC, a significant immunostaining decrease that could not be explained by neuronal loss. However, interneurons expressing parvalbumin and neuropeptide Y were found to be decreased throughout the PC, whereas cholecystokinin-positive cells were diminished in superficial layers. These findings demonstrate synaptic hyperexcitability that is contributed by attenuated inhibition in the PC of pilocarpine-treated epileptic rats and underscore the role of PC networks in temporal lobe epilepsy.

The epileptic human hippocampal cornu ammonis 2 region generates spontaneous interictal-like activity in vitro

The dentate gyrus, the cornu ammonis 2 region and the subiculum of the human hippocampal formation are resistant to the cell loss associated with temporal lobe epilepsy. The subiculum, but not the dentate gyrus, generates interictal-like activity in tissue slices from epileptic patients. In this study, we asked whether a similar population activity is generated in the cornu ammonis 2 region and examined the electrophysiological and neuroanatomical characteristics of human epileptic cornu ammonis 2 neurons that may be involved. Hippocampal slices were prepared from postoperative temporal lobe tissue derived from epileptic patients. Field potentials and multi-unit activity were recorded in vitro using multiple extracellular microelectrodes. Pyramidal cells were characterized in intra-cellular records and were filled with biocytin for subsequent anatomy. Fluorescent immunostaining was made on fixed tissue against the chloride-cation cotransporters sodium-potassium-chloride cotransporter-1 and potassium-chloride cotransporter-2. Light and electron microscopy were used to examine the parvalbumin-positive perisomatic inhibitory network. In 15 of 20 slices, the hippocampal cornu ammonis 2 region generated a spontaneous interictal-like activity, independently of population events in the subiculum. Most cornu ammonis 2 pyramidal cells fired spontaneously. All cells fired single action potentials and burst firing was evoked in three cells. Spontaneous excitatory postsynaptic potentials were recorded in all cells, but hyperpolarizing inhibitory postsynaptic potentials were detected in only 27% of the cells. Two-thirds of cornu ammonis 2 neurons showed depolarizing responses during interictal-like events, while the others were inhibited, according to the current sink in the cell body layer. Two biocytin-filled cells both showed a pyramidal-like morphology with axons projecting to the cornu ammonis 2 and cornu ammonis 3 regions. Expression of sodium-potassium-chloride cotransporter-1 and potassium-chloride cotransporter-2 was reduced in some cells of the epileptic cornu ammonis 2 region, but not to an extent corresponding to the proportion of cells in which hyperpolarizing postsynaptic potentials were absent. Numbers of parvalbumin-positive inhibitory cells and axons were shown to be decreased in the epileptic tissue. Electron microscopy showed the preservation of somatic inhibitory input of cornu ammonis 2 cells, and confirmed the loss of parvalbumin from the interneurons rather than their death. An extra excitatory input (partly coming from sprouted mossy fibres) was demonstrated to innervate cornu ammonis 2 cell bodies. Our results show that the cornu ammonis 2 region of the sclerotic human hippocampus can generate an independent epileptiform activity. Inhibitory and excitatory signalling were functional but modified in epileptic cornu ammonis 2 pyramidal cells. Overexcitation and the altered functional properties of perisomatic inhibitory network, rather than a modified chloride homeostasis, may account for the perturbed gamma-aminobutyric acid-ergic signalling and the generation of interictal-like activity in the human epileptic cornu ammonis 2 region.

Two Distinct Types of Inhibition Mediated by Cartwheel Cells in the Dorsal Cochlear Nucleus

Individual neurons have been shown to exhibit target cell-specific synaptic function in several brain areas. The time course of the postsynaptic conductances (PSCs) strongly influences the dynamics of local neural networks. Cartwheel cells (CWCs) are the most numerous inhibitory interneurons in the dorsal cochlear nucleus (DCN). They are excited by parallel fiber synapses, which carry polysensory information, and in turn inhibit other CWCs and the main projection neurons of the DCN, pyramidal cells (PCs). CWCs have been implicated in "context-dependent" inhibition, producing either depolarizing (other CWCs) or hyperpolarizing (PCs) post synaptic potentials. In the present study, we used paired whole cell recordings to examine target-dependent inhibition from CWCs in neonatal rat DCN slices. We found that CWC inhibitory postsynaptic potentials (IPSPs) onto PCs are large (1.3 mV) and brief (half-width = 11.8 ms), whereas CWC IPSPs onto other CWCs are small (0.2 mV) and slow (half-width = 36.8 ms). Evoked IPSPs between CWCs exhibit paired-pulse facilitation, while CWC IPSPs onto PCs exhibit paired-pulse depression. Perforated-patch recordings showed that spontaneous IPSPs in CWCs are hyperpolarizing at rest with a mean estimated reversal potential of -67 mV. Spontaneous IPSCs were smaller and lasted longer in CWCs than in PCs, suggesting that the kinetics of the receptors are different in the two cell types. These results reveal that CWCs play a dual role in the DCN. The CWC-CWC network interactions are slow and sensitive to the average rate of CWC firing, whereas the CWC-PC network is fast and sensitive to transient changes in CWC firing.

Network hyperexcitability within the deep layers of the pilocarpine‐treated rat entorhinal cortex

In this study we report that in the presence of normal buffer, epileptiform discharges occur spontaneously (duration = 2.60 +/- 0.49 s) or can be induced by electrical stimuli (duration = 2.50 +/- 0.62 s) in the entorhinal cortex (EC) of brain slices obtained from pilocarpine-treated rats but not in those from age-matched, nonepileptic control (NEC) animals. These network-driven epileptiform events consist of field oscillatory sequences at frequencies greater than 200 Hz that most often initiate in the lateral EC and propagate to the medial EC with 4-63 ms delays. The NMDA receptor antagonist CPP depresses the rate of occurrence (P < 0.01) of these spontaneous epileptiform discharges but fails in blocking them. Paradoxically, stimulus-induced epileptiform responses are enhanced in duration during CPP application. However, concomitant application of NMDA and non-NMDA glutamatergic antagonists abolishes spontaneous and stimulus-induced epileptiform events. Intracellular recordings from lateral EC layer V cells indicate a lower frequency of spontaneous hyperpolarizing postsynaptic potentials in pilocarpine-treated tissue than in NEC (P < 0.002) both under control conditions and with glutamatergic receptor blockade; the reversal potential of pharmacologically isolated GABA(A) receptor-mediated inhibitory postsynaptic potentials has similar values in the two types of tissue. Finally, immunohistochemical analysis shows that parvalbumin-positive interneurons are selectively reduced in number in EC deep layers. Collectively, these results indicate that reduced inhibition within the pilocarpine-treated EC layer V may promote network epileptic hyperexcitability.

Ionic mechanism of long-lasting discharges of action potentials triggered by membrane hyperpolarization in the medial lateral habenula.

The activation of inhibitory synapses typically suppresses the generation of action potentials by hyperpolarizing the membrane of postsynaptic cells. In contrast to such conventional action of inhibitory synapses, we report here the ionic mechanism through which hyperpolarizing synapses trigger long-lasting discharges of action potentials that persist up to several tens of seconds. By using extracellular and intracellular recordings in slice preparations, we demonstrate that the activation of synaptic input from the limbic forebrain generates transient hyperpolarizing postsynaptic potentials in neurons of the medial part of the lateral habenular nucleus of the epithalamus. The synaptic hyperpolarization then sets off the coordinated activation of a distinct set of membrane ion channels and intracellular Ca2+ mobilization by internal stores. The activation of these cellular events in distinct temporal order drives a persistent depolarization of habenular cells and promotes long-lasting discharges of tonic action potentials. The cells in the medial division of the lateral habenula project to dopamine and serotonin cells in the midbrain. We suggest that these habenular cells, by generating persistent action potentials in response to a transient increase in the activity of the limbic forebrain, may contribute to the regulation of the serotonergic and dopaminergic activity in the brain.

Two new actions of topiramate: inhibition of depolarizing GABA A-mediated responses and activation of a potassium conductance

Topiramate (TPM) is an antiepileptic with several proposed mechanisms of action including the inhibition of carbonic anhydrase (CA). Since the activity of this enzyme is essential for the generation of GABA A-mediated depolarizing responses, which appears to participate in epileptogenesis, we investigated whether TPM could inhibit such a response in rat hippocampal slices using intracellular recordings. Bath perfusion of TPM (20 and 100 μM) reversibly reduced the GABA A-mediated depolarizing responses evoked by either synaptic stimulation (GDPSPs) or by pressure application of GABA, but did not modify the GABA A-mediated hyperpolarizing postsynaptic potentials. TPM (20 μM) shifted the reversal potential for the GDPSP by −10 mV. Unexpectedly, TPM also induced a steady membrane hyperpolarization associated with a reduction in the input resistance of the cell. This effect was insensitive to tetrodotoxin, and to GABA A and GABA B receptor antagonists, but was blocked by barium (1 mM). Notably, when the extracellular concentration of K + was varied the reversal potential shifted as predicted by the Nernst potential for K +. Acetazolamide (20 μM), another CA inhibitor, elicited similar effects to those reported here for TPM and occluded the hyperpolarization evoked by TPM. The results of this study support the concept that inhibition of carbonic anhydrase in neurons contributes to the anticonvulsant activity of TPM.

Dopamine-Dependent Synaptic Plasticity in the Striatal Cholinergic Interneurons

The striatum, the input stage of the basal ganglia, is a critical brain structure for the learning of stimulus-response habits as well as motor, perceptual, and cognitive skills. Roles of dopamine (DA) and acetylcholine (ACh) in this form of implicit memory have long been considered essential, but the underlying cellular mechanism is still unclear. By means of patch-clamp recordings from corticostriatal slices of the mouse, we studied whether the identified striatal cholinergic interneurons undergo long-term synaptic changes after tetanic stimulation of cortico- and thalamostriatal fibers. Electrical stimulation of the fibers revealed a depolarizing and hyperpolarizing postsynaptic potential in the striatal cholinergic interneurons. The early depolarizing phase was considered to be a cortico/thalamostriatal glutamatergic EPSP, and the hyperpolarizing component was considered to be an intrastriatally evoked GABAergic IPSP. Tetanic stimulation of cortico/thalamostriatal fibers was found to induce simultaneously occurring long-term potentiation (LTP) of the EPSPs as well as the disynaptically mediated IPSPs. The induction of LTP of EPSP required a rise in intracellular Ca(2+) concentration and dopamine D(5), but not D(2) receptor activation. Ca(2+)-permeable AMPA receptors might also play a part in the LTP induction. Blockade of NMDA receptors, metabotropic glutamate receptors, or serotonin receptors had no significant effects. The long-term enhancement of the disynaptic IPSPs was caused by a long-term increase in the occurrence rate but not the amplitude of disynaptically mediated IPSP in the striatal cholinergic interneurons. This dual mechanism of synaptic plasticity may be responsible for the long-term modulation of the cortico/thalamostriatal synaptic transmission.

Potentiation of GABAergic synaptic transmission in the rostral nucleus of the solitary tract

Whole-cell recordings were made from neurons in the rostral nucleus of the solitary tract in horizontal brainstem slices. Monosynaptic GABA A receptor-mediated inhibitory postsynaptic potentials were evoked by single stimulus shocks or by high-frequency tetanic stimulation in the presence of glutamate receptor blockers. While single stimulus-evoked inhibitory postsynaptic potentials had variable amplitudes, tetanic stimulation-induced, hyperpolarizing postsynaptic potentials were of a more constant amplitude. Furthermore, tetanic stimulation resulted in potentiation of the amplitude of single stimulus shock-evoked inhibitory postsynaptic potentials. Of 55 neurons that were tested, potentiation lasted over 30 min for 11, 10–30 min for 13, less than 10 min for 23 and no potentiation occurred in eight. Tetanic stimulation did not result in potentiation of the tetanic stimulus-evoked hyperpolarizing postsynaptic potentials. Both the single stimulus shock- and tetanic stimulus-evoked potentials had similar inhibition concentration–response curves to the GABA A antagonist, bicuculline methiodide (EC 50=0.75 and 0.83, respectively), indicating that they were mediated by the same postsynaptic receptors. By comparing the effect of bicuculline methiodide on the amplitude of the single stimulus shock-evoked inhibitory postsynaptic potentials and the tetanic stimulus-evoked hyperpolarizing potentials, we concluded that a single stimulus shock does not activate all postsynaptic GABA A receptors. However, tetanic stimulation results in activation of all postsynaptic GABA A receptors and induces long-lasting changes in the presynaptic GABAergic neuron. These long-lasting changes of the presynaptic neuron facilitate the release of GABA during single stimulus shock and, as a consequence, more postsynaptic receptors are activated during single stimulus shock-evoked synaptic transmission. This conclusion is supported by the results of experiments in which the extracellular Ca 2+ concentration was manipulated to change the amount of neurotransmitter released from the presynaptic GABAergic terminals. The single stimulus shock-evoked inhibitory postsynaptic potentials were sensitive to the extracellular Ca 2+ concentration, whereas tetanic stimulus-evoked inhibitory postsynaptic potentials were essentially insensitive to extracellular Ca 2+ concentration. The relationship between the single stimulus shock-evoked inhibitory postsynaptic potential amplitude and extracellular Ca 2+ concentration indicates that, in control physiological saline containing 2.5 mM Ca 2+, a single stimulus shock activates less than half the postsynaptic GABA receptors. The phenomenon of long-lasting potentiation of inhibitory transmission within the rostral nucleus of the solitary tract may be important in the processing of gustatory information and play a role in taste-guided behaviors.

RGS proteins reconstitute the rapid gating kinetics of gbetagamma-activated inwardly rectifying K+ channels.

G protein-gated inward rectifier K+ (GIRK) channels mediate hyperpolarizing postsynaptic potentials in the nervous system and in the heart during activation of Galpha(i/o)-coupled receptors. In neurons and cardiac atrial cells the time course for receptor-mediated GIRK current deactivation is 20-40 times faster than that observed in heterologous systems expressing cloned receptors and GIRK channels, suggesting that an additional component(s) is required to confer the rapid kinetic properties of the native transduction pathway. We report here that heterologous expression of "regulators of G protein signaling" (RGS proteins), along with cloned G protein-coupled receptors and GIRK channels, reconstitutes the temporal properties of the native receptor --> GIRK signal transduction pathway. GIRK current waveforms evoked by agonist activation of muscarinic m2 receptors or serotonin 1A receptors were dramatically accelerated by coexpression of either RGS1, RGS3, or RGS4, but not RGS2. For the brain-expressed RGS4 isoform, neither the current amplitude nor the steady-state agonist dose-response relationship was significantly affected by RGS expression, although the agonist-independent "basal" GIRK current was suppressed by approximately 40%. Because GIRK activation and deactivation kinetics are the limiting rates for the onset and termination of "slow" postsynaptic inhibitory currents in neurons and atrial cells, RGS proteins may play crucial roles in the timing of information transfer within the brain and to peripheral tissues.

Synaptic interactions due to convergent input from gustatory afferent fibers in the rostral nucleus of the solitary tract.

1. Whole cell recordings from neurons in the rostral, gustatory nucleus of the solitary tract (rNST) were made using the "blind" patch-clamp technique in horizontal brain stem slices of rats. 2. Postsynaptic potentials (PSP) were elicited in 71 rNST neurons by electrical stimulation of the solitary tract (ST). To investigate PSPs evoked by convergent input from the chorda tympani and glossopharyngeal nerves, the ST was stimulated at levels where these two nerves terminate. These are referred to as rostral (rST) and intermediate (iST) ST, respectively. 3. When the rST was stimulated 72% of the PSPs were depolarizing, and 28% were hyperpolarizing (n = 64). Stimulation of the intermediate ST resulted in 75% depolarizing and 25% hyperpolarizing PSPs (n = 56). 4. Application of gamma-aminobuturic acid-A (GABAA) and glutamate receptor blockers revealed that all PSPs recorded in the present study were a composite of summed excitatory and inhibitory PSPs. Application of the GABAA receptor blocker bicuculline, by eliminating the hyperpolarizing component of a PSP, revealed the excitatory postsynaptic potential (EPSP) component of the potential. Bicuculline also increased the amplitude and prolonged the decay time of the depolarizing potentials once the hyperpolarizing potential component had been eliminated. These pure EPSP revealed by GABAA receptor blockade reversed at approximately 0 mV. 5. Application of glutamate ionotropic receptor blockers effectively eliminated the initiation of the synaptic responses evoked by ST stimulation. If the stimulus strength was increased, an inhibitory postsynaptic potential (IPSP) was elicited, presumably by direct activation of interneurons close to the stimulating electrode. These IPSPs had a mean reversal potential of -88 mV. 6. When synaptic responses were initiated by stimulation of the projection areas of both the chorda tympani and glossopharyngeal nerves, all neurons tested (n = 49) responded to stimulation of both sites on the ST. The resulting synaptic potential was a sum of the two individual synaptic potentials. 7. If stimulation of the rostral and intermediate sites both elicited depolarizing potentials, the potential resulting from stimulation of both sites was the arithmetical sum of the two individual PSPs. The EPSPs summed even if the time between stimulation of the rostral and intermediate sites was separated by < or = 100 ms. 8. Inhibitory PSPs evoked by simultaneous stimulation of the rostral and intermediate ST also summed. The summation was not linear and saturated at a mean level of -66 mV. 9. When the PSP at one stimulation site was excitatory but inhibitory at the other site, the PSP wave form resulting from dual stimulation was a complex mixture of the two individual potentials. The inhibitory potential was capable of blocking action potentials resulting from the excitatory PSP. 10. These results indicate that synaptic responses in rNST are complex mixtures of excitatory and inhibitory potentials. The synaptic potentials result from excitatory afferent input mediated by glutamate and inhibitory input derived from interneurons. Inhibitory transmission is mediated primarily at GABAA synapses. Stimulation of the rNST afferent input derived from the chorda tympani and glossopharyngeal nerves reveals complex convergent input. The complexity of these synaptic interactions indicates that considerable processing of gustatory information occurs at the first central synapse in the taste pathway.

Extrinsic Inhibitory Innervation to Rubral Neurons in Rat Brain-Stem Slices

Synaptic connections between the neurons in the red nucleus (RN) and its extrinsic neurons were studied using rat brain-stem slices. Intracellular records were obtained from the RN neurons. Ipsilateral stimuli to areas in the dorsolateral mesencephalic reticular formation (DLMRF) or substantia nigra (SN) elicited monosynaptic hyperpolarizing postsynaptic potentials (PSPs) in about 95% of RN neurons recorded. The hyperpolarizing PSPs could be reversibly blocked by bicuculline, indicating that they were GABAAreceptor-mediated–Cl−-inhibitory PSPs. The sites of most inhibitory synapses arising from DLMRF and SN are possibly located on the proximal half of the soma–dendritic membrane of RN neurons, according to the analysis of the IPSPs with Rall’s model. In addition, tracing dyes were employed to examine the morphological pathways. After rhodamine B, a retrograde tracer, was applied to the RN in brain slices, the cell bodies of a number of neurons in DLMRF and SN were labeled. These labeled neurons were also immunopositive for glutamic acid decarboxylase (GAD) as revealed from double labeling with an anti-GAD antiserum. The anterograde tracer, tetramethylrhodamine dextran, was applied to the DLMRF or SN and taken up by many neurons in the areas. A portion of these cells extended their processes toward and terminated within the RN. Moreover, electron microscopic examination confirmed that the tetramethylrhodamine dextran-decorated synaptic terminals were present in the RN. The results indicate that the rubral neurons receive direct GABAAreceptor-mediated inhibitory inputs from neurons in the DLMRF and SN, which may participate in modulation of rubral outputs.

Local inhibitory synaptic inputs to neurones of the paraventricular nucleus in slices of rat hypothalamus.

1. Intracellular recordings were obtained from neurones in the region of the paraventricular nucleus in slices of rat hypothalamus. Glutamate microdrops were applied to the surface of the slices at sites dorsal, lateral and ventral to the paraventricular nucleus to selectively activate local presynaptic neurones. The gamma-aminobutyric acidA (GABAA)-receptor antagonists picrotoxin or bicuculline were bath-applied to block synaptic inhibition. 2. Glutamate microapplication caused a tonic depolarization and often repetitive action potentials in twenty of forty-seven recorded cells. This was probably caused by the direct exposure of the dendrites of the recorded cells to the glutamate microdrops. 3. Glutamate microstimulation elicited inhibitory synaptic responses in nine of forty-seven neurones tested. Glutamate microdrops caused discrete, hyperpolarizing postsynaptic potentials (PSPs) in four cells recorded with microelectrodes containing potassium acetate and evoked depolarizing PSPs in four cells recorded with KCl-filled microelectrodes. Glutamate microapplication inhibited spontaneous spike firing in another cell recorded with a potassium acetate microelectrode. 4. Bath application of GABAA-receptor antagonists completely blocked the hyperpolarizing PSPs elicited by glutamate microstimulation in three of three cells recorded with potassium acetate electrodes and the depolarizing PSPs in two of two cells recorded with KCl electrodes, indicating they were inhibitory PSPs caused by the release of GABA. Suppression of GABAA-mediated synaptic inhibition did not reveal any glutamate-evoked excitatory PSPs. 5. Recorded cells were identified as magnocellular, parvocellular or non-paraventricular bursting neurones on the basis of their electrophysiological properties. Direct depolarization and local inhibitory synaptic responses were observed in all three cell types. 6. Several conclusions can be drawn from these data: (1) functional glutamate receptors are distributed throughout neuronal populations in the paraventricular region of the hypothalamus, confirming and extending previous observations; (2) local synaptic inputs to neurones in the paraventricular nucleus are primarily inhibitory, supplied by perinuclear GABAergic neurones; (3) both magnocellular and parvocellular subpopulations receive local inhibitory synaptic inputs. The possibility that these local GABAergic circuits mediate inhibitory inputs to paraventricular neurones from limbic structures is discussed.

Synaptic responses of guinea pig cingulate cortical neurons in vitro

1. Intracellular recordings were made from layer V/VI neurons of the guinea pig anterior cingulate cortex to investigate postsynaptic potentials (PSPs) evoked by electrical stimulation of the subcortical white matter (forceps minor). 2. Four distinct types of PSPs were recorded (at the resting potential) under normal physiological conditions; 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX)-sensitive excitatory postsynaptic potentials (EPSPs) were followed by bicuculline- or picrotoxin-sensitive depolarizing or hyperpolarizing inhibitory postsynaptic potentials (IPSPs), which were further followed by phaclofen-sensitive, long-lasting hyperpolarizing postsynaptic potentials (LPSPs). The average times-to-peak for the EPSP, depolarizing and hyperpolarizing IPSPs, and LPSP were 10, 22, 28, and 146 ms, respectively. 3. In the presence of CNQX and bicuculline, high-intensity electrical stimulation elicited a longer lasting EPSP with a time-to-peak of 21 ms. The amplitude and duration of the EPSP decreased with membrane hyperpolarization and increased with membrane depolarization. The EPSP was reversibly abolished by D,L-2-amino-5-phosphonovaleric acid (D,L-APV). 4. The bicuculline- or picrotoxin-sensitive depolarizing and hyperpolarizing IPSPs and the phaclofen-sensitive LPSP were markedly suppressed by CNQX, suggesting that glutamate (Glu) and/or aspartate nerve terminals project to GABAergic interneurons, and that the GABAergic interneurons are activated mainly by non-N-methyl-D-aspartate (non-NMDA) receptors. 5. In the presence of picrotoxin, the average reversal potential for the compound EPSP was 0 mV, which was similar to that (-6 mV) for the Glu-induced depolarization. In a solution containing D,L-APV at low concentrations, the average reversal potentials for the depolarizing and hyperpolarizing IPSPs and for the early and late components of the gamma-aminobutyric acid (GABA)-induced responses were -62, -72, -70, and -61 mV, respectively. Thus the value for the depolarizing IPSP was similar to that for the late response to GABA, whereas the value for the hyperpolarizing IPSP was almost the same as that for the early response to GABA. The average reversal potential of -90 mV for the LPSP was similar to -93 mV for the baclofen-induced hyperpolarization and to -94 mV for the spike afterhyperpolarization. 6. Application of phaclofen decreased the interspike interval of the spontaneous firing and reversed the increase in the interspike interval after subcortical stimulation. This result indicates that, even in a slice preparation, the anterior cingulate neurons are under tonic inhibitory control exerted by spontaneously active GABAergic interneurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenosine inhibits a GABAB receptor-mediated hyperpolarizing postsynaptic potential in neurons of rat septal nuclei.

Intracellular recordings were made from neurons of rat dorsolateral septal nucleus (DLSN), in vitro. Adenosine and 2-chloroadenosine (1-500 microM) hyperpolarized DLSN neurons and blocked the excitatory postsynaptic potential (EPSP) and the late hyperpolarizing potential (LHP) in the presence of bicuculline. Adenosine did not depress the glutamate-induced potential. Bath-application of adenosine depressed the baclofen-induced potential in 60% of the neurons. Adenosine also inhibited the LHP in the remaining 40% of neurons, while it did not depress the baclofen-induced potential in these neurons. These results indicate that adenosine inhibits the EPSP pre-synaptically whereas it inhibits the LHP both pre- and postsynaptically in rat septal nuclei.

Inhibitory postsynaptic potentials in neonatal rat sympathetic preganglionic neurones in vitro.

1. Intracellular recordings were made from antidromically identified sympathetic preganglionic neurones (SPNs) in transverse sections of thoraco-lumbar spinal cord from neonatal (12-22 day) rats. 2. Two types of hyperpolarizing (inhibitory) postsynaptic potentials (IPSPs) were recorded in the SPNs. The first type, which we have termed unitary IPSPs, were small, discrete IPSPs that occurred spontaneously and also following chemical or electrical stimulation applied to the spinal cord slices. The second type IPSP was a hyperpolarizing response evoked by either dorsal or ventral root stimulation. 3. Spontaneously occurring unitary IPSPs had an amplitude of 1 to 5 mV, and reversal potential of -60 to -75 mV; they were reversibly abolished by low Ca2+, tetrodotoxin (TTX) or strychnine but not by bicuculline and picrotoxin. 4. Pressure application of N-methyl-D-aspartate (NMDA), an excitatory amino SPNs; these were abolished by either strychnine or by the NMDA receptor antagonist D-2-amino-5-phosphonovalerate. Furthermore, electrical stimulation of dorsal rootlets elicited in several SPNs the discharge of strychnine-sensitive unitary IPSPs. 5. Electrical stimulation applied to dorsal or ventral rootlets elicited in nineteen and eight SPNs, respectively, an IPSP of larger amplitude (5 to 15 mV). The IPSP exhibited a reversal potential of -60 to 75 mV; it was changed to a depolarizing response in a low [Cl-]o solution, but was not significantly affected in a low [K+]o. Strychnine but not bicuculline or picrotoxin reversibly blocked the IPSPs in nearly all the SPNs. Additionally, hexamethonium and d-tubocurarine antagonized the IPSPs evoked by ventral but not by dorsal root stimulations. 6. Our results suggest that unitary and evoked IPSPs recorded in SPNs are due primarily to an increase of Cl- conductance by glycine or a glycine-like substance, released from interneurones, that can be activated by NMDA. Furthermore, IPSPs evoked by ventral root stimulation appear to represent a disynaptic event whereby nicotinic activation of a glycine-releasing interneurone results in a release of the inhibitory transmitter; this is then analogous to the Renshaw cell circuitry of the spinal motoneurones.

Neuromodulatory action of dopamine in the nucleus accumbens: An in vivo intracellular study

Intracellular recordings were made from neurons in the nucleus accumbens in situ to determine how dopamine produces the selective neuromodulatory action in the accumbens observed in previous studies. Electrical stimulation of the basolateral nucleus of the amygdala was found to produce monosynaptically evoked depolarizing and hyperpolarizing postsynaptic potential sequences in a large proportion of the accumbens neurons sampled. Dopamine applied iontophoretically or released endogenously by stimulation of the ventral tegmental area produced consistent membrane depolarization and an increase in membrane conductance but not an increase in spontaneous activity of the accumbens neurons. Stimulation of the ventral tegmental area with trains of 10 pulses at 10 Hz prior to stimulation of the amygdala produced 8–58% reduction in the amplitude of the depolarizing postsynaptic potential but no change in the late hyperpolarizing postsynaptic potential. Although attenuation of the depolarizing postsynaptic potential amplitude from ventral tegmental area stimulation was often accompanied by membrane depolarization, it appeared that the two responses were not causally related. The effect of ventral tegmental area stimulation on the evoked depolarizing postsynaptic potential and the membrane potential were blocked by haloperidol indicating the involvement of dopamine. Iontophoretically applied dopamine produced responses similar to ventral tegmental area stimulation with two exceptions: (i) iontophoretically applied dopamine produced consistently stronger maximal attenuation of the depolarizing postsynaptic potential than did ventral tegmental area stimulation; and (ii) iontophoretically applied dopamine always attenuated both the depolarizing postsynaptic potential and hyperpolarizing postsynaptic potential whereas ventral tegmental area stimulation produced selective attenuation of the depolarizing postsynaptic potential only. These electrophysiological results are complementary to those from pharmacological experiments and suggest that one of several physiological functions of dopamine in the nucleus accumbens is a neuromodulatory one involving presynaptic action on non-dopaminergic terminals.

Repetitive synaptic potentials responsible for inhibition of spinal cord motoneurons during active sleep

High-gain intracellular recordings of spinal cord motoneurons were obtained in the chronically prepared cat during wakefulness, quiet sleep, and active sleep. Discrete hyperpolarizing postsynaptic potentials were found to occur repetitively throughout active sleep in conjunction with the presence of somatomotor atonia. These potentials were most prominent in the transition period from quiet to active sleep as well as during episodes of rapid eye movements during active sleep. Their polarity was reversed after the intracellular injection of chloride ions. We propose that these are inhibitory potentials which are responsible for or contribute significantly to the postsynaptic inhibition of spinal cord motoneurons during active sleep.

Postsynaptic potentials in pacemaker cells: a correlation of behavior in command cells of an electric fish.

Intracellular recordings were made from pacemaker-command cells of the electric organ discharge (EOD) of the weakly electric fish Eigenmannia virescens. The fish was immobilized with gallamine triethiodide (Flaxedil) which silenced the EOD. A simulated EOD of this fish (ca. 300 Hz) and a sine wave simulating a neighbor, a few Hz higher (+deltaF) or lower (-deltaF) were introduced into the bath to elicit the "jamming avoidance response" (JAR), monitored through the pacemaker potential. We observed that accompanying the JAR there is a minute hyperpolarizing postsynaptic potential (hpsp) superimposed on the pacemaker potential. A shift in the phase of the hpsp occurs with a change in the sign of deltaF, and therefore of the JAR. Assuming that the behaviorally correlated hpsp is inhibitory, it suggests that mutual inhibition may play a role in regulating the synchronous firing frequency of command neurons, which are electrically coupled with one-another. Scheich and Bullock (1974) proposed a neuronal scheme for the JAR in which they suggest that two systems (P and T) operate together in the nervous system. The T system affects the pacemaker cells at a precise, variable phase of the pacemaker cycle. Although the present results indeed reveal a shift in the hpsp with a change in the sign of deltaF, the actual significance of this shift remains to be evaluated. The unexpected direction of the shift suggests either that the hpsp is excitatory at the phases when it occurs, or that effectiveness of inhibition decreases at later phases in this case instead of increasing as in other cases, or that the hpsp opposes the JAR. The parallel P system is probably more important in explaining the JAR, acting by a DC level control rather than a phase control.

Inhibition of hypoglossal motoneurons by stimulation of the jaw-opening muscle afferents.

The synaptic linkage of jaw-opening muscle afferents with the retractive and the protrusive motoneurons (R-Mns and P-Mns) in the hypoglossal motor nucleus was explored in anesthetized cats. It was found that stimulation of low-threshold afferents arising in the jaw-opening muscles (m. mylohyoideus and m. digastricus) evoked IPSPs in both R-Mns and P-Mns with about 10 to 15 msec latencies. By injecting Cl ions into R-Mns or P-Mns, the hyperpolarizing postsynaptic potentials evoked by jaw-opening muscle afferent impulses were reversed to depolarizing potentials. The lingual-IPSPs of hypoglossal motoneurons were concluded to be dependent on increase of Cl conductance. Stimulation of low threshold lingual afferents evoked a reflex discharge in the retractive nerve fibers with a 6.0 msec latency and in the protrusive nerve fibers with a 5.5 msec latency. Inhibition of the linguohypoglossal reflex by stimulation of the jaw-opening muscle afferents was completely removed by an intravenous injection of strychnine (0.1 mg/kg) or picrotoxin (6 mg/kg). It is concluded that the low threshold afferents arising in the jaw-opening muscles are connected to both R-Mns and P-Mns and P-Mns through inhibitory synapses via a polysynaptic pathway.