GABA(B) receptors inhibit backpropagating dendritic spikes in hippocampal CA1 pyramidal cells in vivo

Spike backpropagation has been proposed to enhance dendritic depolarization and synaptic plasticity. However, relatively little is known about the inhibitory control of spike backpropagation in vivo. In this study, the backpropagation of the antidromic spike into the dendrites of CA1 pyramidal cells was studied by extracellular recording in urethane-anesthetized rats. The population antidromic spike (pAS) in CA1 following stimulation of the alveus was recorded simultaneously with a 16-channel silicon probe and analyzed as current source density (CSD). The pAS current sink was shown to sequentially invade the soma and then the apical and basal dendrites. When the pAS was preceded <400 ms by a conditioning orthodromic CA3 stimulus, the apical and basal dendritic spike sinks were reduced and delayed. Dendritic spike suppression was large after a high-intensity CA3 conditioning stimulus that evoked a population spike, small after a low-intensity CA3 conditioning stimulus, and weak after conditioning by another pAS. The late (150-400 ms latency) inhibition of the backpropagating pAS at the apical and basal dendrites was partially relieved by a GABA(B) receptor antagonist, CGP35348 or CGP56999A, given intracerebroventricularly (icv). CGP35348 icv also decreased the latency of the antidromic spike sinks at all depths. A compartment cable model of a CA1 pyramidal cell with excitable dendrites, combined with a model of extracellular potential generation, confirms that GABA(B) receptor activation delays a backpropagating spike and blocks distal dendritic spikes. GABA(B) receptor-mediated conductance increase and hyperpolarization, amplified by removing dendritic I(A) inactivation, contribute to conditioned dendritic spike suppression. In addition, the model shows that slow Na(+) channel inactivation also participates in conditioned spike suppression, which may partly explain the small dendritic spike suppression after conditioning with a weak orthodromic stimulus or another antidromic spike. Thus, both theory and experiment confirm an important role of the GABA(B) receptors in controlling dendritic spike backpropagation.     

Mechanisms of long-term potentiation: a current-source density analysis

A current-source density (CSD) analysis was carried out in the CA 1 region of the hippocampal slice (1) to determine the pattern of current flow in pyramidal cells upon orthodromic stimulation and (2) to test the hypothesis that EPSP-to-spike potentiation is produced by an alteration in this distribution of current sinks and/or sources. The results indicated that 2 sinks occur near the cell body layer (in addition to the sink associated with the EPSP) in response to orthodromic stimulation of the apical dendrites. An early (i.e., short-latency) sink was present along the radiatum/pyramidale border and was evident throughout the time course of the evoked field potential. This sink peaked in magnitude just prior to the peak of the population spike and was associated with orthodromic stimulation; it was not seen with antidromic stimulation. A second, later, sink occurred in the proximal portion of the basal dendrites and had a characteristic time course similar to the population spike; this second sink was also present during antidromic stimulation. There was some suggestion that the earlier dendritic sink shifted apically with development of long-term potentiation (LTP). The existence and movement of such an active zone in these cells may help to explain the dissociation of EPSP and spike potentiation in LTP.     

Argiotoxin-636 blocks excitatory synaptic transmission in rat hippocampal CA1 pyramidal neurons

Argiotoxin 636, (AR₆₃₆), a synaptic antagonist from orb weaver spider venom, is shown produce reversible blockade of excitatory transmission in CA1 pyramidal neurons of the in vitro rat hippocampus. Microtopical applicationof AR₆₃₆ (5–50 nM) resulted in a concentration-dependent suppression of the amplitude of the dendritic field EPSP recorded from stratum radiatum, and the amplitude of the population spike recorded from stratum pyramidale in response to stimulation of the Schaffer collaterals. The maximum effect of AR₆₃₆ occured at about 15–25 min. These effects were reversible after washing with toxin-free physiological solution with the rate of recovery having an inverse relationships to the concnetration of AR₆₃₆. In contrast to the effects observed with orthodromic stimulation, the amplitude of the antidromic spike was not affected by exposure to AR₆₃₆. The temporal pattern GABAergic paired-pulse inhibition was unaffected by exposure to AR₆₃₆. Neuronal discharge elicited by pressure injection of -glutamate was abolished by AR₆₃₆, whereas, responses to -aspartate were not significantlu affected. These data suggest that AR₆₃₆ functionsas a selective antagonist of glutamate-mediated synaptic transmission in rat hippocampus.     

Electrophysiologic study of non-pyramidal neurons of the stratum radiatum in slices of guinea pig hippocampus

The activity of 67 nonpyramidal neurons of str. radiatum-moleculare (NSRM) and of 8 presumed interneurons of str. oriens-pyramiidale (NSOP) was recorded extracellularly in guinea pig hippocampal slices. In comparison with high frequency grouped discharges characteristic of NSOP, NSRM had low frequency background activity consisting of single (77%) and grouped (23%) spikes. The level of the background activity of NSRM decreased with an increase of the distance of their location from str. pyramidale. Electrical stimulation of dentate fascia usually evoked 1-2 spike discharges in NSRM, while bursts of spikes were evoked in NSOP. The thresholds of responses in NSRM were not different or higher than those of pyramidal neurons, while in NSOP they were significantly lower. The period of suppression of the spontaneous activity usually followed the evoked spike discharge in NSRM. During evoked synchronous epileptiform discharges of pyramidal neurons the simultaneous excitation of NSRM was observed. Excitatory influence of pyramidal neurons on NSRM and participation of the latter in dendritic inhibition of pyramidal neurons is suggested.     

The sources of extracellular potassium accumulation in the CA1 region of hippocampal slices

To estimate the relative contributions of pre- and postsynaptic elements, and of synaptic and action potential-related currents, to the elevation of interstitial potassium ([K+]o) that occurs during neural activation, we measured [K+]o and focal electrical potential (Vec) during ortho- and antidromic stimulation, before and after blocking synaptic transmission, in the CA1 region of hippocampal tissue slices in vitro. Single stimulus pulses could cause delta [K+]o as large as 0.25 mM in stratum (st.) pyramidale and 0.27 mM in st. radiatum; stimulus trains could cause delta [K+]o as large as 10.5 mM in st. pyramidale and 6.25 mM in st. radiatum. Stimulus trains also caused negative Vec shifts; these shifts were related in a linear fashion to delta [K+]o. For a given increase in [K+]o, the change in Vec was greater in st. radiatum than in st. pyramidale. In st. pyramidale, the delta [K+]o evoked by antidromic stimulation was 65% of the delta [K+]o evoked by orthodromic stimulation (with equal population spike amplitudes). Blockade of synaptic transmission by removal of Ca2+ reduced orthodromically evoked delta [K+]o in st. radiatum by 52%; delta [K+]o in st. pyramidale was abolished. Removal of Ca2+ caused an 11% decrease in the delta [K+]o evoked in st. pyramidale by antidromic stimulation. We conclude that in the layer of dendritic trees (st. radiatum), approximately half of the K+ ions released into the interstitial space during orthodromic stimulation come from presynaptic terminals, with the remainder probably resulting from the ion currents of postsynaptic potentials. Among the pyramidal cell bodies (st. pyramidale), almost all of the excess K+ is released by action potential currents.     

Developmental changes in neuronal sensitivity to excitatory amino acids in area CA1 of the rat hippocampus

The laminar distribution of decreases in extracellular free calcium and concomitant field potentials induced by repetitive orthodromic stimulation, ionophoretic application of N-methyl-D-aspartate and quisqualate, was studied in the CA1 field of rat hippocampal slices, at two different stages during postnatal development. While stimulation-elicited and quisqualate-induced signals remain maximal in stratum pyramidale during the first postnatal month, the laminar profiles of responses to N-methyl-D-aspartate (NMDA) depend on age: the responses to this agonist are maximal in stratum pyramidale in 5-9-day-old rats and in stratum radiatum in 12-30-day-old rats. Our findings suggest that, during the second postnatal week, the apical dendrites of pyramidal neurons in area CAl become more sensitive to NMDA, which is expressed by big influxes of calcium at this level.     

A hypoxic injury potential in the hippocampal slice

In rat hippocampal slices, neurons in the stratum pyramidale of the CA1 were stimulated orthodromically and antidromically while the resultant extracellular population spikes were monitored. Hypoxic conditions were then induced. After disappearance of the orthodromic population spike, a second orthodromic population spike appeared. We have titled this the hypoxic injury potential since it reflects the onset of permanent injury to neurons in area CA1 of the hippocampus.     

The effects of antidromic discharges on orthodromic firing of primary afferents in the cat

This study investigated the effects of antidromically conducted nerve impulses on the transmission of orthodromic volleys in primary afferents of the hindlimb in decerebrated paralyzed cats. Two protocols were used: (A) Single skin and muscle afferents (N=20) isolated from the distal part of cut dorsal rootlets (L7-S1) were recorded while stimulation was applied more caudally. The results showed that during the trains of three to 20 stimuli, the orthodromic firing frequency decreased or ceased, depending on the frequency of stimulation. Remarkably, subsequent to these trains, the occurrence of orthodromic spikes could be delayed for hundreds of ms (15/20 afferents) and sometimes stopped for several seconds (10/20 afferents). Longer stimulation trains, simulating antidromic bursts reported during locomotion, caused a progressive decrease, and a slow recovery of, orthodromic firing frequency (7/20 afferents), indicating a cumulative long-lasting depressing effect from successive bursts. (B) Identified stretch-sensitive muscle afferents were recorded intra-axonally and antidromic spikes were evoked by the injection of square pulses of current through the micropipette. In this case, one to three antidromic spikes were sufficient to delay the occurrence of the next orthodromic spike by more than one control inter-spike interval. If the control inter-spike interval was decreased by stretching the muscle, the delay evoked by antidromic spikes decreased proportionally. Overall, these findings suggest that antidromic activity could alter the mechanisms underlying spike generation in peripheral sensory receptors and modify the orthodromic discharges of afferents during locomotion.     

The establishment of GABAergic and glutamatergic synapses on CA1 pyramidal neurons is sequential and correlates with the development of the apical dendrite.

We have performed a morphofunctional analysis of CA1 pyramidal neurons at birth to examine the sequence of formation of GABAergic and glutamatergic postsynaptic currents (PSCs) and to determine their relation to the dendritic arborization of pyramidal neurons. We report that at birth pyramidal neurons are heterogeneous. Three stages of development can be identified: (1) the majority of the neurons (80%) have small somata, an anlage of apical dendrite, and neither spontaneous nor evoked PSCs; (2) 10% of the neurons have a small apical dendrite restricted to the stratum radiatum and PSCs mediated only by GABA(A) receptors; and (3) 10% of the neurons have an apical dendrite that reaches the stratum lacunosum moleculare and PSCs mediated both by GABA(A) and glutamate receptors. These three groups of pyramidal neurons can be differentiated by their capacitance (C(m) = 17.9 +/- 0.8; 30.2 +/- 1.6; 43.2 +/- 3.0 pF, respectively). At birth, the synaptic markers synapsin-1 and synaptophysin labeling are present in dendritic layers but not in the stratum pyramidale, suggesting that GABAergic peridendritic synapses are established before perisomatic ones. The present observations demonstrate that GABAergic and glutamatergic synapses are established sequentially with GABAergic synapses being established first most likely on the apical dendrites of the principal neurons. We propose that different sets of conditions are required for the establishment of functional GABA and glutamate synapses, the latter necessitating more developed neurons that have apical dendrites that reach the lacunosum moleculare region.     

Alterations of inhibitory synaptic responses in the dentate gyrus of temporal lobe epileptic patients

The number of orthodromically evoked population spikes was used to classify brain slice tissue from the dentate gyrus of temporal lobe epileptic patients as “more excitable” (multiple population spikes) or “less excitable” (a single population spike). During orthodromic stimulation, “more excitable” tissue exhibited less paired-pulse depression in comparison to “less excitable” tissue. During antidromic stimulaltion, both multiple population spikes and paired-pulse depression were observed in “more excitable” tissue. “Less excitable” tissue exhibited a single antidromic spike and often on antidromically evoked paired-pulse depression. The strength of antidromic paired-pulse depression was correlated positively with the number of antidromic spikes and was correlated negatively with orthodromic paired-pulse depression. Although orthodromic and antidromic paired-pulse depression were correlated to the number of orthodromically evoked populaltion spikes, this correlation was not as strong as that between orthodromic paired-pulse depression, antidromic paired-pulse depression, and number of antidromically evoked population spikes. The antidromic paired-pulse depression observed in tissue exhibiting antidromically evoked multiple population spikes was enhanced rather than blocked by bicuculline. In addition, the blockade of the antidromic paired-pulse depression by CNQX indicated that this inhibition is mediated by an AMPA-type glutamatergic synapse. We suggest that alterations in circuitry occur in the dentate gyrus of some temporal lobe epileptic patients and were manifested by both a loss of inhibitory input as well as an increase of inhibition, which was dependent on the pathway of stimulaltion. The results of pairing antidromic and orthodromic stimuli were consistent with these conclusions. © 1994 Wiley-Liss, Inc.     

Extracellular potassium ion activity and electrophysiology in the hippocampal slice: paradoxical recovery of synaptic transmission during anoxia

The relationship between extracellular potassium ion activity and neuronal excitability during anoxia was investigated in hippocampal slices in vitro. Extracellular field potentials and K+ activity were measured with double-barreled ion-selective microelectrodes placed either in the stratum pyramidale or stratum radiatum of field CA1. Orthodromic spike activity of CA1 pyramidal cells and field excitatory postsynaptic potentials (f-EPSPs) failed rapidly after anoxia with little change in potassium ion activity and without failure of the Schaffer collateral prevolley or antidromic responses of pyramidal cells. As [K+]o approached 8-10 mM, f-EPSPs and orthodromic spike activity recovered spontaneously. Continued anoxia resulted in massive release of K+ into the extracellular space and complete electrical silence. Presynaptic activity and antidromically elicited spike activity recovered promptly upon reoxygenation after anoxia, but synaptic transmission remained blocked for many minutes. Spontaneous recovery of f-EPSPs and spike activity suggests that a simple mechanism involving depolarization or hyperpolarization of neuronal elements cannot account for failure of synaptic transmission observed during anoxia. However, continued elevation of [K+]o and the associated loss of pre- and postsynaptic excitability with more prolonged anoxia indicated that depolarization was responsible for the eventual electrical silence as anoxia progressed.     

Synaptic control of excitability in isolated dendrites of hippocampal neurons

The apical dendrites of CA1 pyramidal cells were isolated from their cell bodies by making cuts through proximal stratum radiatum of transverse hippocampal slices from the guinea pig. This lesion separated the distal apical dendritic elements from the somata, basal dendrites, and 50 to 100 microns of the proximal apical dendritic tree. Orthodromic stimuli in stratum radiatum evoked excitatory synaptic responses in isolated dendrites, but no phasic inhibitory components could be detected. In spite of this surgically produced disinhibition, orthodromic stimuli did not elicit burst activity at the resting membrane potential. However, isolated dendrites and intact dendrites could generate multiple slow spike activity when directly stimulated with depolarizing current pulses. When isolated dendrites were depolarized by DC current, excitatory postsynaptic potentials could evoke subthreshold intrinsic slow depolarizations, or repetitive slow spikes, similar to responses elicited by depolarizing current pulses alone. After exposure to bicuculline (5 microns), both intact and isolated dendrites generated bursts of activity following synaptic activation. A possible mechanism for this action of bicuculline is blockade of a residual GABA-mediated inhibition which was not expressed as a postsynaptic hyperpolarization in isolated dendrites. This bicuculline-sensitive event was capable of depressing dendritic excitability in the absence of the recurrent inhibitory synaptic input and was very effective in controlling burst activity. Our results indicate that the dendritic electrical behavior is dependent on a complex interaction between synaptic and voltage-sensitive events.     

Spontaneous EEG spikes in the normal hippocampus. I. Behavioral correlates, laminar profiles and bilateral synchrony

Spontaneous EEG and unit activities were recorded from the CA1 region of the dorsal hippocampus by means of a movable microelectrode in normal behaving rats. Large amplitude (less than 4 mV) negative EEG spikes (SPKs) of 40-100 msec duration with frequencies in the range of 0.2-5/sec were consistently recorded from the middle apical dendritic layer (stratum radiatum) during awake immobility, grooming and slow-wave sleep. SPKs were replaced by rhythmical slow activity (RSA) during walking and paradoxical sleep. Laminar analysis indicated that SPKs were positive in stratum oriens, negative in stratum radiatum and polarity reversal just below stratum pyramidale. Peak positivity (about 1 mV on average) and peak negativity (2 mV) occurred some 80 micron above and 200 micron below the reversal point, respectively. The SPKs were invariably accompanied by synchronous burst discharges in stratum pyramidale. Bilateral recordings demonstrated the SPKs occurred synchronously in large areas of the CA1 field of the two hippocampi. These results suggest that the SPK represents a massive synaptic excitation of middle apical dendrites triggering synchronous burst discharges in a population of pyramidal cell bodies. A possibility was discussed that these non-pathological SPKs and interictal spikes share some common underlying mechanisms.     

Stratum lacunosum-moleculare interneurons of hippocampal CA1 region. II. Intrasomatic and intradendritic recordings of local circuit synaptic interactions

Simultaneous intracellular recordings were obtained from stratum lacunosum-moleculare (L-M) interneurons and CA1 cells, and their local circuit synaptic interactions were examined. Synaptic interactions with pyramidal cells were evaluated in both intrasomatic and intradendritic pyramidal cell recordings. Stimulation of L-M interneurons evoked small-amplitude IPSPs in 21% of intrasomatic (9/42 cell pairs) and in 26% of intradendritic (11/43) pyramidal cell recordings. The IPSP mean peak amplitude was 0.91 mV for intrasomatic and 0.67 mV for intradendritic recordings. IPSPs had slow onset and decay (approximately 80-90 msec), decreased in amplitude with membrane hyperpolarization, and were not associated with any apparent change in input resistance. No physiologic evidence of synaptic connections was found from pyramidal cells to L-M interneurons. Inhibitory synaptic interactions were also seen between L-M interneurons and stratum pyramidale interneurons (2 of 4 cell pairs). The IPSPs recorded in pyramidale interneurons were similar to the IPSPs recorded in pyramidal cells. During simultaneous recordings, L-M interneurons were activated at a shorter latency, i.e., in a feedforward manner with respect to pyramidal cells. Thus, L-M interneurons may mediate feedforward inhibition of CA1 pyramidal cells. The L-M interneuron-evoked IPSPs in pyramidal cells share some characteristics of the late IPSP recorded in CA1 pyramidal cells and may therefore contribute to this component of the IPSP.     

A specific effect of morphine on evoked activity in the rat hippocampal slice

The effect of morphine (0.5-50 microM) was examined on CA1 field potentials in the tranverse hippocampal slice. Morphine consistently produced an augmentation of evoked activity manifest as (i) a decrease in the threshold for generation of a population spike and (ii) generation of an additional population spike(s) whose amplitude was proportional to the position of the sampled response on its input/output curve. Both of these opiate effects were stereospecific and naloxone-reversible. Additional population spikes occurred in opiate medium with either orthodromic or antidromic activation of the pyramidal cells, and the antidromic effect was abolished when synaptic transmission was blocked, suggesting that morphine did not act directly upon the pyramidal cells. Recordings of population EPSPs in the dendrites of the pyramidal cells showed no changes due to opiate exposure near threshold. Opiate effects were mimicked by the gamma-aminobutyric acid (GABA) antagonist picrotoxin, and were partially to fully reversed by GABA itself, suggesting that disinhibition of pyramidal cells might be involved as a mechanism in this opiate effect. The data are evidence for a specific primary effect of morphine within the hippocampus in spite of the low numbers of opiate receptors in this brain region.     

A new aspect of the collision test principle: cell body distance from recording site

A theoretical method is described for estimating the distance between a spike recording-site, possibly axonal, and the corresponding cell body of unknown location. The method requires that an orthodromic spike be recorded following an antidromic spike, with estimation of a collision interval analogous to that used for establishing antidromicity. To calculate the distance between recording-site and cell body, values are needed for the collision interval between antidromic and succeeding orthodromic spikes, the refractory period of the spike, and the antidromic conduction speed. Problems may arise in determining the last value. The method is illustrated with antidromic spikes recorded in the medial thalamus of the cat upon stimulating the caudate nucleus.     

Increased proximal bifurcation of CA1 pyramidal apical dendrites in sema3A mutant mice

Semaphorin‐3A (Sema3A) is an attractive guidance molecule for cortical apical dendrites. To elucidate the role of Sema3A in hippocampal dendritic formation, we examined the Sema3A expression pattern in the perinatal hippocampal formation and analyzed hippocampal dendrites of the brains from young adult sema3A mutant mice. Sema3A protein was predominantly expressed in the hippocampal plate and the inner marginal zone at the initial period of apical dendritic growth. Neuropilin‐1 and plexin‐A, the receptor components for Sema3A, were also localized in the same regions. The Golgi impregnation method revealed that in wildtype mice more than 90% of hippocampal CA1 pyramidal neurons extended a single trunk or apical trunks bifurcated in stratum radiatum. Seven percent of the pyramidal neurons showed proximal bifurcation of apical trunks in stratum pyramidale or at the border of the stratum pyramidale and stratum radiatum. In sema3A mutant mice, proximally bifurcated apical dendrites were increased to 32%, while the single apical dendritic pyramidal neurons were decreased. We designate this phenotype in sema3A mutant mice as “proximal bifurcation.” In the dissociated culture system, approximately half of the hippocampal neurons from wildtype mice resembled pyramidal neurons, which possess a long, thick, and tapered dendrite. In contrast, only 30% of the neurons from sema3A mutants exhibited pyramidal‐like morphology. Proximal bifurcation of CA1 pyramidal neurons was also increased in the mutant mice of p35, an activator of cyclin‐dependent kinase 5 (Cdk5). Thus, Sema3A may facilitate the initial growth of CA1 apical dendrites via the activation of p35/Cdk5, which may in turn signal hippocampal development. J. Comp. Neurol. 516:360–375, 2009. © 2009 Wiley‐Liss, Inc.     

Burst generating and regular spiking layer 5 pyramidal neurons of rat neocortex have different morphological features

Intracellular recordings were obtained from pyramidal neurons in layer 5 of rat somatosensory and visual cortical slices maintained in vitro. When directly depolarized, one subclass of pyramidal neurons had the capacity to generate intrinsic burst discharges and another generated regular trains of single spikes. Burst responses were triggered in an all-or-none manner from depolarizing afterpotentials in most bursting neurons. Regular spiking cells responded to electrical stimulation of ascending afferents with a typical EPSP-IPSP sequence, whereas IPSPs were hard to detect in bursting cells. Orthodromic activation of the latter evoked a prominent voltage-dependent depolarization that could trigger a burst response. Intracellularly labelled bursting and regular spiking cells were located in layer 5b, but had distinctly different morphologies. Bursting neurons had a large pyramidal soma, a gradually emerging apical dendrite, and an extensive apical and basal dendritic tree. Their axonal collateral arborization was predominantly limited to layers 5/6. In contrast, regular spiking cells had a more rounded soma with abruptly emerging apical dendrite, a smaller dendritic arborization, and 2 to 8 ascending axonal collaterals that arborized widely in the supragranular layers. Both bursting and regular spiking cells had main axons that entered the subcortical white matter. These data show that some subgroups of pyramidal neurons within the deeper parts of layer 5 of rat cortex are morphologically and physiologically distinct and have different intracortical connections. Bursting cells presumably function to amplify and synchronize cortical outputs, whereas regular spiking output neurons provide excitatory feedback to neurons at all cortical levels and receive a more effective orthodromic inhibitory input. These data support the hypothesis that differences in gross neuronal structure, perhaps even the subtle differences that distinguish subclasses of neurons in a given lamina, are predictive of underlying differences in the type and distribution of ion channels in the nerve cell membrane and connections of cells within the cortical circuit.     

Post-ischemic potentiation of Schaffer collateral/CA1 pyramidal cell responses of the rat hippocampus in vivo: involvement of receptors

The present study examined the functional changes in the hippocampal CA1 pyramidal cell system in vivo occurring after 12-min forebrain ischemia in the rat. A population excitatory postsynaptic potential and orthodromic population spike of CA1 pyramidal cells to stimulation of the Schaffer collaterals were potentiated at 6–8 h post-ischemia. These changes were not associated with an increase in excitability of the CA1 pyramidal cells as evaluated from the antidromic population spike induced by alveus stimulation, suggesting the presence of an increased synaptic efficacy. The post-ischemic potentiation was prevented by pretreatment with the (NMDA) receptor antagonist, MK801, in a dose-dependent manner. These findings suggest that 12-min forebrain ischemia in the rat activates NMDA receptors, which results in an increase in synaptic efficacy to the CA1 pyramidal cells at 6–8 h post-ischemia.     

Chronic ethanol treatment reduces inhibition in CA1 of the rat hippocampus

The effect of chronic ethanol exposure on inhibition in the rat hippocampal slice was investigated using paired-pulse stimulation techniques with stimulation in stratum radiatum or stratum oriens of CA1. Experimental animals were fed ethanol in a liquid diet for 20 weeks and were withdrawn for at least 8 weeks prior to electrophysiological recording. Prior ethanol treatment had no effect on basic input-output relationships for the extracellular population spike. Ethanol treatment significantly reduced the recurrent inhibition produced by antidromic stimulation in a manner dependent upon stimulus intensity. In addition, with orthodromic paired-pulse stimulation of either stratum radiatum or oriens, a trend toward an augmentation of the facilitation of population spike amplitude was observed, suggesting that feedforward inhibition may also be reduced. These results are similar to those found with treatments that reduce inhibition. Therefore, we conclude that chronic ethanol exposure produces an enduring disruption of inhibitory neuronal function in the rat hippocampus.