Fast Inhibitory Postsynaptic Potentials Research Articles

P 188. Pharmacological characterization of TMS-evoked EEG responses by positive modulators at the GABA-A receptor

Introduction The combined use of Transcranial Magnetic Stimulation (TMS) and Electroencephalography (EEG) enables a direct investigation of cortical-evoked responses in humans. The physiology of the peaks characterizing the TMS-evoked EEG responses has not been clearly elucidated yet. EEG is able to detect fast-inhibitory post synaptic potentials (fIPSPs, Objectives In the present study we characterized the pharmaco-physiology of TEPs in healthy volunteers by using two GABA-A receptors positive modulators: alprazolam and zolpidem. The main aim of the study was to test to which extent early TEPs are affected by fast inhibitory neurotransmission through the GABA-A receptor. Materials and methods Twenty healthy subjects participated in a pseudorandomized, placebo-controlled, double-blind crossover design, using a single oral dose of alprazolam (1 mg), a classical benzodiazepine preferentially binding on alpha1, alpha2, alpha3 and alpha5 subunit bearing subtypes of the GABA-A receptor, and zolpidem (10 mg), which mainly binds at the alpha1 subtype. TEPs were recorded by TMS-EEG before and 90 min after drug administration and the effects of the drugs on the TEP amplitudes were statistically evaluated. Results A non parametric cluster-based permutation analysis of amplitudes at channel-level was run for placebo, alprazolam and zolpidem administration. Both alprazolam and zolpidem, increased the amplitude of the negative potential at 45 ms after stimulation (N45). Moreover, a strong positive correlation was found between their modulation on the N45 magnitude. In addition, alprazolam but not zolpidem decreased the amplitudes of the later TEPs, in particular, the negative at 100 ms (N100) and the positive at 180 ms (P180). Conclusion We provide first-time evidence that the N45 potential may reflect a biological marker of alpha1 containing GABA-A receptor activity. The broader modulation of several TEP components by alprazolam could be explained by its wider pharmacological profile, as it is responsible for the activation of several subtypes of GABA-A receptors.

Dynamic causal models and physiological inference: a validation study using isoflurane anaesthesia in rodents.

Generative models of neuroimaging and electrophysiological data present new opportunities for accessing hidden or latent brain states. Dynamic causal modeling (DCM) uses Bayesian model inversion and selection to infer the synaptic mechanisms underlying empirically observed brain responses. DCM for electrophysiological data, in particular, aims to estimate the relative strength of synaptic transmission at different cell types and via specific neurotransmitters. Here, we report a DCM validation study concerning inference on excitatory and inhibitory synaptic transmission, using different doses of a volatile anaesthetic agent (isoflurane) to parametrically modify excitatory and inhibitory synaptic processing while recording local field potentials (LFPs) from primary auditory cortex (A1) and the posterior auditory field (PAF) in the auditory belt region in rodents. We test whether DCM can infer, from the LFP measurements, the expected drug-induced changes in synaptic transmission mediated via fast ionotropic receptors; i.e., excitatory (glutamatergic) AMPA and inhibitory GABAA receptors. Cross- and auto-spectra from the two regions were used to optimise three DCMs based on biologically plausible neural mass models and specific network architectures. Consistent with known extrinsic connectivity patterns in sensory hierarchies, we found that a model comprising forward connections from A1 to PAF and backward connections from PAF to A1 outperformed a model with forward connections from PAF to A1 and backward connections from A1 to PAF and a model with reciprocal lateral connections. The parameter estimates from the most plausible model indicated that the amplitude of fast glutamatergic excitatory postsynaptic potentials (EPSPs) and inhibitory postsynaptic potentials (IPSPs) behaved as predicted by previous neurophysiological studies. Specifically, with increasing levels of anaesthesia, glutamatergic EPSPs decreased linearly, whereas fast GABAergic IPSPs displayed a nonlinear (saturating) increase. The consistency of our model-based in vivo results with experimental in vitro results lends further validity to the capacity of DCM to infer on synaptic processes using macroscopic neurophysiological data.

Spike-timing dependent plasticity in inhibitory circuits

Inhibitory circuits in the brain rely on GABA-releasing interneurons. For long, inhibitory circuits were considered weakly plastic in the face of patterns of neuronal activity that trigger long-term changes in the synapses between excitatory principal cells. Recent studies however have shown that GABAergic circuits undergo various forms of long-term plasticity. For the purpose of this review, we identify three major long-term plasticity expression sites. The first locus is the glutamatergic synapses that excite GABAergic inhibitory cells and drive their activity. Such synapses, on many but not all inhibitory interneurons, exhibit long-term potentiation (LTP) and depression (LTD). Second, GABAergic synapses themselves can undergo changes in GABA release probability or postsynaptic GABA receptors. The third site of plasticity is in the postsynaptic anion gradient of GABAergic synapses; coincident firing of GABAergic axons and postsynaptic neurons can cause a long-lasting change in the reversal potential of GABAA receptors mediating fast inhibitory postsynaptic potentials. We review the recent literature on these forms of plasticity by asking how they may be triggered by specific patterns of pre- and postsynaptic action potentials, although very few studies have directly examined spike-timing dependent plasticity (STDP) protocols in inhibitory circuits. Plasticity of interneuron recruitment and of GABAergic signaling provides for a rich flexibility in inhibition that may be central to many aspects of brain function. We do not consider plasticity at glutamatergic synapses on Purkinje cells and other GABAergic principal cells.

Learning-induced enhancement of feedback inhibitory synaptic transmission

Olfactory-discrimination learning results with a series of intrinsic and excitatory synaptic modifications in piriform cortex pyramidal neurons. Here we show that such learning results with long-lasting enhancement of inhibitory synaptic transmission onto proximal dendrites of these pyramidal neurons. Such enhancement is mediated by a strong hyperpolarizing shift in the reversal potential of fast inhibitory postsynaptic potentials (fIPSPs). Moreover, paired-pulse depression of these IPSPs, indicating enhanced GABA release, is also apparent after learning. We suggest that learning is accompanied by long-lasting enhancement of synaptic inhibition onto excitatory neurons, thus compensating for the increase of excitation in these neurons.

Synaptic 5 Subunit-Containing GABAA Receptors Mediate IPSPs Elicited by Dendrite-Preferring Cells in Rat Neocortex

Previous studies indicated that one class of dendrite-preferring hippocampal interneurones inhibits pyramidal cells via alpha 5 gamma-aminobutyric acid (GABA(A)) receptors whereas parvalbumin- and CCK-containing basket cells act via alpha1 and alpha2/3 GABA(A) receptors, respectively. This study asked whether there is selective insertion of different alpha subunit-containing GABA(A) receptors at neocortical inhibitory synapses innervated by specific classes of interneurones. The benzodiazepine site pharmacology of inhibitory postsynaptic potentials (IPSPs) elicited in neocortical pyramidal cells by 3 classes of interneurones was explored with dual whole-cell recordings in neocortical slices from juvenile rats (P18-23). Fast IPSPs activated by multipolar interneurones with narrow spikes and nonadapting firing patterns were powerfully enhanced by the alpha1-preferring agonist zolpidem, suggesting mediation via larger proportion of alpha1 GABA(A) receptors than those activated by multipolar, adapting interneurones, which were less strongly enhanced by zolpidem, but equally insensitive to the alpha 5-selective inverse agonist IA alpha 5 (MSD, Essex, UK) suggesting mediation predominantly via alpha2/3 GABA(A) receptors. In contrast, the IPSPs elicited by bitufted, dendrite-preferring interneurones were reduced by IA alpha 5 and by zinc and insensitive to zolpidem despite enhancement by the broad-spectrum agonist, diazepam. Thus insertion of GABA(A) receptors at synapses on neocortical pyramids is input-specific, with proximal inhibition employing alpha1 and alpha2/3 GABA(A) receptors and dendrite-preferring bitufted interneurones activating alpha 5 GABA(A) receptors.

Dopamine Presynaptically Depresses Fast Inhibitory Synaptic Transmission via D4 Receptor-Protein Kinase A Pathway in the Rat Dorsolateral Septal Nucleus

The lateral septal nucleus receives a diffuse dopaminergic input originating from the ventral tegmental area of the brain stem. We examined whether dopamine (DA) modulates synaptic transmission in the slice preparation of the rat dorsolateral septal nucleus (DLSN). Bath application (10-15 min) of DA (30 muM) markedly depressed the amplitude of fast and slow inhibitory postsynaptic potentials (IPSPs) in DLSN neurons, while it produced only a minor depression of the amplitude of excitatory postsynaptic potentials (EPSPs) obtained in the presence of bicuculline. DA (30 muM) depressed the monosynaptic fast IPSP to approximately 50% of control, but did not depress the inward current (I(GABA)) induced by exogenous gamma-aminobutyric acid (GABA). DA decreased the frequency of miniature fast IPSPs (m-fIPSPs) without significantly changing their amplitude. PD 168077, a selective D4 receptor agonist, depressed the fast and slow IPSPs but not the EPSP and decreased the frequency of m-fIPSPs. Both DA and PD 168077 increased the paired-pulse ratio of the monosynaptic fast IPSP. The inhibitory effect of DA on the fast IPSP was significantly attenuated by L-741,742, an antagonist at D4 receptors, but not by SCH 23390 and sulpiride, a D1-like and a D2-like receptor antagonist, respectively. N-ethylmaleimide, a blocker of pertussis toxin (PTX)-sensitive G protein (G(i/o)), attenuated the DA-induced depression of the fast IPSP. N-[2-((p-bromocinnamyl) amino)ethyl]-5-isoquinoline sulfonamide, a protein kinase A (PKA) inhibitor, attenuated the DA-induced depression of the fast IPSP. These results suggest that DA inhibits spontaneous and evoked release of GABA via the D4 receptor-G(i)-protein-PKA system in DLSN neurons.

Developmental changes in presynaptic muscarinic modulation of excitatory and inhibitory neurotransmission in rat piriform cortex in vitro: Relevance to epileptiform bursting susceptibility

Suppression of depolarizing postsynaptic potentials and isolated GABA-A receptor-mediated fast inhibitory postsynaptic potentials by the muscarinic acetylcholine receptor agonist, oxotremorine-M (10 μM), was investigated in adult and immature (P14-P30) rat piriform cortical (PC) slices using intracellular recording. Depolarizing postsynaptic potentials evoked by layers II–III stimulation underwent concentration-dependent inhibition in oxotremorine-M that was most likely presynaptic and M2 muscarinic acetylcholine receptor-mediated in immature, but M1-mediated in adult (P40-P80) slices; percentage inhibition was smaller in immature than in adult piriform cortex. In contrast, compared with adults, layer Ia-evoked depolarizing postsynaptic potentials in immature piriform cortex slices in oxotremorine-M, showed a prolonged multiphasic depolarization with superimposed fast transients and spikes, and an increased ‘all-or-nothing’ character. Isolated N-methyl- d-aspartate receptor-mediated layer Ia depolarizing postsynaptic potentials (although significantly larger in immature slices) were however, unaffected by oxotremorine-M, but blocked by dl-2-amino-5-phosphonovaleric acid. Fast inhibitory postsynaptic potentials evoked by layer Ib or layers II–III-fiber stimulation in immature slices were significantly smaller than in adults, despite similar estimated mean reversal potentials (∼−69 and −70 mV respectively). In oxotremorine-M, only layer Ib-fast inhibitory postsynaptic potentials were suppressed; suppression was again most likely presynaptic M2-mediated in immature slices, but M1-mediated in adults. The degree of fast inhibitory postsynaptic potential suppression was however, greater in immature than in adult piriform cortex. Our results demonstrate some important physiological and pharmacological differences between excitatory and inhibitory synaptic systems in adult and immature piriform cortex that could contribute toward the increased susceptibility of this region to muscarinic agonist-induced epileptiform activity in immature brain slices.

Altered Inhibition in Lateral Amygdala Networks in a Rat Model of Temporal Lobe Epilepsy

Clinical and experimental evidence indicates that the amygdala is involved in limbic seizures observed in patients with temporal lobe epilepsy. Here, we used simultaneous field and intracellular recordings from horizontal brain slices obtained from pilocarpine-treated rats and age-matched nonepileptic controls (NECs) to shed light on the electrophysiological changes that occur within the lateral nucleus (LA) of the amygdala. No significant differences in LA neuronal intrinsic properties were observed between pilocarpine-treated and NEC tissue. However, spontaneous field activity could be recorded in the LA of 21% of pilocarpine-treated slices but never from NECs. At the intracellular level, this network activity was characterized by robust neuronal firing and was abolished by glutamatergic antagonists. In addition, we could identify in all pilocarpine-treated LA neurons: 1) large amplitude depolarizing postsynaptic potentials (PSPs) and 2) a lower incidence of spontaneous hyperpolarizing PSPs as compared with NECs. Single-shock stimulation of LA networks in the presence of glutamatergic antagonists revealed a biphasic inhibitory PSP (IPSP) in both NECs and pilocarpine-treated tissue. The reversal potential of the early GABA(A) receptor-mediated component, but not of the late GABA(B) receptor-mediated component, was significantly more depolarized in pilocarpine-treated slices. Furthermore, the peak conductance of both fast and late IPSP components had significantly lower values in pilocarpine-treated LA cells. Finally, paired-pulse stimulation protocols in the presence of glutamatergic antagonists revealed a less pronounced depression of the second IPSP in pilocarpine-treated slices compared with NECs. Altogether, these findings suggest that alterations in both pre- and postsynaptic inhibitory mechanisms contribute to synaptic hyperexcitability of LA networks in epileptic rats.

Neural Mechanisms of the Ultrafast Activities

A brief review of previous studies is presented on ultra-fast activities > 300 Hz (high frequency oscillations, HFOs) overlying the cortical response in the somatosensory evoked potential (SEP) or magnetic field (SEF). The characteristics of somatosensory HFOs are described in terms of reproducibility and origin (area 3b and 1) of the HFOs, changes during a wake-sleep cycle, effects of higher stimulus rate or tactile interference, etc. Also, several hypotheses on the neural mechanisms of the HFOs are introduced; the early HFO burst is probably generated from action potentials of thalamocortical fibers at the time when they arrive at the area 3b (and 1), since this component is resistant to higher stimulus rate > 10Hz or general anesthesia: by contrast, the late HFO burst is sensitive to higher stimulus rate, reflecting activities of a postsynaptic neural network in the somatosensory cortices, area 3b and 1. As to possible mechanisms of the late HFO burst genesis, an interneuron hypothesis, a fast inhibitory postsynaptic potential (IPSP) hypothesis of the pyramidal cell and a chattering cell hypothesis will be discussed on the basis of physiological and pathological features of the somatosensory HFOs.

Synaptic hyperexcitability of deep layer neocortical cells in a genetic model of absence seizures

We used sharp-electrode, intracellular recordings in an in vitro brain slice preparation to study the excitability of neocortical neurons located in the deep layers (>900 microm from the pia) of epileptic (180-210-days old) Wistar Albino Glaxo/Rijswijk (WAG/Rij) and age-matched, non-epileptic control (NEC) rats. Wistar Albino Glaxo/Rijswijk rats represent a genetic model of absence seizures associated with generalized spike and wave (SW) discharges in vivo. When filled with neurobiotin, these neurons had a typical pyramidal shape with extensive apical and basal dendritic trees; moreover, WAG/Rij and NEC cells had similar fundamental electrophysiological and repetitive firing properties. Sequences of excitatory postsynaptic potentials (EPSPs) and hyperpolarizing inhibitory postsynaptic potentials (IPSPs) were induced in both the strains by electrical stimuli delivered to the underlying white matter or within the neocortex; however, in 24 of 55 regularly firing WAG/Rij cells but only in 2 of 25 NEC neurons, we identified a late EPSP that (1) led to action potential discharge and (2) was abolished by the N-methyl-D-aspartate (NMDA) receptor antagonist 3,3-(2-carboxypiperazine-4-yl)-propyl-1-phosphonate (20 microM; n = 8/8 WAG/Rij cells). Finally, we found that the fast and slow components of the stimulus-induced IPSPs recorded during the application of glutamatergic receptor antagonists had similar reversal potentials in the two strains, while the peak conductance of the fast IPSP was significantly reduced in WAG/Rij cells. These findings document an increase in synaptic excitability that is mediated by NMDA receptors, in epileptic WAG/Rij rat neurons located in neocortical deep layers. We propose that this mechanism may be instrumental for initiating and maintaining generalized SW discharges in vivo.

Weaker synaptic inhibition in CA1 region of ventral compared to dorsal rat hippocampal slices

Extracellular and intracellular recordings were made from slices taken from the dorsal (DH) and ventral (VH) part of rat hippocampus. Using paired-pulse stimulation of Schaffer collaterals, at different interpulse intervals (IPIs), and records of the population spike (PS) we found that the strength and duration of paired-pulse inhibition was much weaker in VH compared to DH slices: at the IPI of 10 ms the decrease of PS in VH (40%) was significantly smaller compared to that in DH slices (76%), while at 20 ms the decrease of PS in DH slices (60%) corresponded to facilitation in VH slices. Moreover, the amplitude and duration of intracellularly recorded fast inhibitory postsynaptic potentials (fast-IPSPs) were found significantly smaller in VH (5.2±0.6 mV, 54.8±5.8 ms) than in DH (11.2±1.1 mV, 105±10 ms) neurons. The smaller and shorter fast-IPSP recorded in VH neurons may at least in part explain the results in paired-pulse inhibition. The demonstrated weaker inhibition may underlie the higher propensity of the ventral hippocampus for epileptiform activity.

Does the Development of a GABAergic Phenotype by Hippocampal Dentate Gyrus Granule Cells Contribute to Epileptogenesis.

Does the Development of a GABAergic Phenotype by Hippocampal Dentate Gyrus Granule Cells Contribute to Epileptogenesis.

Activation of inhibitory pathways suppresses the induction of long-term potentiation in neurons of the rat lateral septal nucleus

Long-term potentiation of the hippocampal-septal pathway was examined by intracellular recording techniques. High frequency stimulation (two 100-Hz 1-s trains with a 20-s interval between them) of the hippocampal CA3 area resulted in a transient depolarization in rat lateral septal nucleus neurons. High frequency stimulation was followed by a facilitation of fast and slow inhibitory postsynaptic potentials, lasting for more than 2 h, but not by a long-lasting increase in the excitatory postsynaptic potential in the normal solution. Long-term potentiation (>2 h) of the excitatory postsynaptic potential did not appear in 74% of neurons tested, even when the fast inhibitory postsynaptic potential was blocked by bicuculline (30 μM), a GABA A receptor antagonist. High frequency stimulation produced long-term potentiation of the excitatory postsynaptic potential in the Mg 2+-free solution containing bicuculline. When the fast and slow inhibitory postsynaptic potentials were blocked by GABA A and GABA B receptor antagonists (bicuculline and CGP 55845A respectively), high frequency stimulation produced a large and sustained depolarization followed by long-term potentiation of the excitatory postsynaptic potential. However, the excitatory postsynaptic potential was not enhanced by administration of these drugs after termination of high frequency stimulation. Pretreatment with 2-amino-5-phosphonopentanoate, a NMDA receptor antagonist, resulted in loss of long-term potentiation in both sets of experiments. Paired-pulse stimulation of the hippocampal CA3 region with interstimulus intervals between 200 and 800 ms depressed the second excitatory postsynaptic potential in the presence of bicuculline. CGP 35348, a GABA B receptor antagonist, reversed the depression of excitatory postsynaptic potentials to facilitation. These data suggest that high frequency stimulation of hippocampal CA3 neurons enhances the efficacy of GABAergic inhibitory circuits which, in turn, depress the ability of lateral septal nucleus neurons to express long-term potentiation.

Kindling induces transient fast inhibition in the dentate gyrus--CA3 projection.

The granule cells of the dentate gyrus (DG) send a strong glutamatergic projection, the mossy fibre tract, toward the hippocampal CA3 field, where it excites pyramidal cells and neighbouring inhibitory interneurons. Despite their excitatory nature, granule cells contain small amounts of GAD (glutamate decarboxylase), the main synthetic enzyme for the inhibitory transmitter GABA. Chronic temporal lobe epilepsy results in transient upregulation of GAD and GABA in granule cells, giving rise to the speculation that following overexcitation, mossy fibres exert an inhibitory effect by release of GABA. We therefore stimulated the DG and recorded synaptic potentials from CA3 pyramidal cells in brain slices from kindled and control rats. In both preparations, DG stimulation caused excitatory postsynaptic potential (EPSP)/inhibitory postsynaptic potential (IPSP) sequences. These potentials could be completely blocked by glutamate receptor antagonists in control rats, while in the kindled rats, a bicuculline-sensitive fast IPSP remained, with an onset latency similar to that of the control EPSP. Interestingly, this IPSP disappeared 1 month after the last seizure. When synaptic responses were evoked by high-frequency stimulation, EPSPs in normal rats readily summate to evoke action potentials. In slices from kindled rats, a summation of IPSPs overrides that of the EPSPs and reduces the probability of evoking action potentials. Our data show for the first time that kindling induces functionally relevant activity-dependent expression of fast inhibition onto pyramidal cells, coming from the DG, that can limit CA3 excitation in a frequency-dependent manner.

Slow inhibitory potentials in the teleost Mauthner cell

In vivo recordings from Mauthner cells in adult zebrafish ( Danio rerio) and goldfish ( Carassius auratus) preparations with potassium chloride filled electrodes revealed a new class of long-lasting synaptic events in these cells. Their decay time constant ranged from 20 to 80 ms, which is about 20 times longer than that of previously identified fast glycinergic inhibitory postsynaptic potentials in this neuron. The average time to peak of these slow events ranged from 1 to 6 ms. We demonstrated that they are also inhibitory since (i) they were resistant to antagonists of the excitatory glutamatergic receptors; (ii) their amplitude was increased following chloride loading of the Mauthner cell; (iii) their reversal potential was the same as that of fast, glycinergic inhibitory postsynaptic potentials; and (iv) they produced an inhibitory shunt of the cell's membrane resistance. Furthermore, as with the fast inhibitory postsynaptic potentials, the decay time of the slow events is voltage dependent, increasing when the Mauthner cell is depolarized. However, these inhibitory postsynaptic potentials had a different pharmacological profile to the fast glycinergic ones. That is, they persisted in the presence of strychnine at doses that abolished the fast ones and they were more sensitive to bicuculline. These data are compatible with the notion that these inhibitory postsynaptic potentials are mediated by activation of a different inhibitory receptor type, and may be GABAergic. In addition, the decay time constant of the fast inhibitory postsynaptic current was shorter than the first of the two components that contribute to the bi-exponential decay reported previously for miniature inhibitory postsynaptic currents in Mauthner cells of larval zebrafish. This suggests developmental modifications and/or a switch in the assembly of glycine receptor subtypes. While amplitude distributions of the fast miniature inhibitory postsynaptic potentials recorded in the presence of tetrodotoxin generally could fit with a single Gaussian function, the amplitude histograms of slow miniature events were skewed, often with multiple nearly equally spaced peaks, consistent with the synchronous release of several quantal units. These previously undescribed slow unitary inhibitory postsynaptic potentials contribute to inhibitory synaptic noise recorded in the Mauthner cells. Specifically, autocorrelation analysis revealed gamma-like rhythms (30–80 Hz) in each of two phases, characterized as “noisy” and “quiet”, and dominated by the fast and slow inhibitory postsynaptic potentials, respectively. The major frequencies of these two states were significantly different (i.e. around 90 and 40 Hz, respectively), suggesting that the fast and slow inhibitory postsynaptic potentials are derived from different inhibitory networks. Chloride-filled Mauthner cells gradually hyperpolarized in the presence of tetrodotoxin, reflecting the effect of ongoing activity in the interneurons that produce the slow events. We conclude that this new class of inhibitory postsynaptic potentials contributes to the tonic inhibition which controls the Mauthner cell's excitability. In physiological conditions, this regulatory influence is expressed as a continuous shunt of this neuron's input resistance and responsiveness to sensory inputs.

Seizures induce simultaneous GABAergic and glutamatergic transmission in the dentate gyrus-CA3 system.

Monosynaptic and polysynaptic responses of CA3 pyramidal cells (PC) to stimulation of the dentate gyrus (DG) are normally blocked by glutamate receptor antagonists (GluRAs). However, after kindled seizures, GluRAs block the monosynaptic excitatory postsynaptic potential (EPSP) and isolate a monosynaptic inhibitory postsynaptic potential (IPSP), suggesting that mossy fibers release GABA. However, kindling epilepsy induces neuronal sprouting, which can underlie this fast inhibitory response. To explore this possibility, the synaptic responses of PC to DG stimulation were analyzed in kindled epileptic rats, with and without seizures, and in nonepileptic rats, immediately after a single pentylenetetrazol (PTZ)-induced seizure, in which sprouting is unlikely to have occurred. Excitatory and inhibitory synaptic responses of PC to DG stimulation were blocked by GluRAs in control cells and in cells from kindled nonseizing rats, confirming that inhibitory potentials are disynaptically mediated. However, a fast IPSP could be evoked in kindled epileptic rats and in nonepileptic rats after a single PTZ-induced seizure. The same response was induced after rekindling the epileptic nonseizing rats. This IPSP has an onset latency that parallels that of the control EPSP and is not altered under low Ca(2+) medium or halothane perfusion. In addition, it was reversibly depressed by L(+)-2-amino-4-phosphonobutyric acid (L-AP4), which is known to inhibit transmitter release from mossy fibers. These results demonstrate that seizures, and not the synaptic rearrangement due to an underlying epileptic state, induce the emergence of fast inhibition in the DG-CA3 system, and suggest that the mossy fibers underlie this plastic change.

Modulatory effects of adenosine on inhibitory postsynaptic potentials in the lateral amygdala of the rat.

1. Adenosine is a depressant in the central nervous system with pre- and postsynaptic effects. In the present study, intracellular recording techniques were applied to investigate the modulatory effects of adenosine on projection neurons in the lateral rat amygdala (LA), maintained as slices in vitro. 2. Adenosine reversibly reduced the amplitude of a fast inhibitory postsynaptic potential (IPSP) that was evoked by electrical stimulation of the external capsule and pharmacologically isolated by applying an N-methyl-D-aspartate and non-N-methyl-D-aspartate receptor antagonist, DL-(-)-2-amino-5-methyl-4-isoxazolepropionic acid and 6, 7-Dinitroquinoxaline-2,3-dione, respectively, and the gamma-aminobutyric acidB (GABAB) receptor antagonist CGP 35348. The postsynaptic potential that remained was abolished by locally applying bicuculline. 3. Adenosine reduced the amplitude of the fast IPSP on average by 40.3%. It had no significant effect on responses to exogenously applied GABA, on membrane potential or on input resistance, suggesting that the site of action was at presynaptic inhibitory interneurons in the LA. 4. The response to adenosine was mimicked by the selective adenosine A1 receptor agonist N6-cyclohexyladenosine and blocked by the selective adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine. 5. Neuronal responsiveness in the amygdala is largely controlled by inhibitory processes. Adenosine can presynaptically downregulate inhibitory postsynaptic responses and could exert dampening effects likely by depression of both excitatory and inhibitory neurotransmitter release.

Recent advances in the pharmacology of quaternary salts of bicuculline.

GABA is the major inhibitory transmitter in the mammalian adult CNS where it acts mainly on GABAA and GABAB receptors. GABAA receptors are heteromultimeric complexes comprising probably five subunits that form a Cl−-permeable channel. These subunits belong to α, β, γ, δ and ϵ families and co-assemble with a variable stoichiometry 1 Sieghart W. Pharmacol. Rev. 1995; 47: 181-234 PubMed Google Scholar , 2 McKernan R.H. Whiting P.J. Trends Neurosci. 1996; 19: 139-143 Abstract Full Text PDF PubMed Scopus (1073) Google Scholar . In addition to the site where GABA binds, most of these receptors also contain binding sites for benzodiazepines and neurosteroids, as well as alcohol and volatile anaesthetics 3 Macdonald R.L. Olsen R.W. Annu. Rev. Neurosci. 1994; 17: 569-602 Crossref PubMed Scopus (1781) Google Scholar , 4 Franks N.P. Lieb W.R. Nature. 1994; 367: 607-614 Crossref PubMed Scopus (1625) Google Scholar , 5 Mihic S.J. et al. Nature. 1997; 389: 385-389 Crossref PubMed Scopus (1096) Google Scholar . Activation of GABAA receptors leads to fast inhibitory postsynaptic potentials (IPSPs) in most brain areas. Because they are often located close to the site where action potentials are generated, these receptors constitute a very strong inhibitory system.

Dual intracellular recordings and computational models of slow inhibitory postsynaptic potentials in rat neocortical and hippocampal slices

Dual intracellular recordings in slices of adult rat neocortex and hippocampus investigated slow, putative GABA B receptor-mediated inhibitory postsynaptic potentials. In most pairs tested in which the interneuron elicited a fast inhibitory postsynaptic potential in the pyramid, this GABA A receptor mediated inhibitory postsynaptic potential was entirely blocked by bicuculline or picrotoxin (3:3 in neocortex, 6:8 in CA1, all CA1 basket cells), even when high-frequency presynaptic spike trains were elicited. However, in three of 85 neocortical paired recordings involving an interneuron, although no discernible response was elicited by single presynaptic interneuronal spikes, a long latency (≥20 ms) inhibitory postsynaptic potential was elicited by a train of ≥3 spikes at frequencies ≥50–100 Hz. This slow inhibitory postsynaptic potential was insensitive to bicuculline (one pair tested). In neocortex, slow inhibitory postsynaptic potential duration reached a maximum of 200 ms even with prolonged presynaptic spike trains. In contrast, summing fast, GABA A inhibitory postsynaptic potentials, elicited by spike trains, lasted as long as the train. Between four and 10 presynaptic spikes, mean peak slow inhibitory postsynaptic potential amplitude increased sharply to 0.38, 2.6 and 2.9 mV, respectively, in the three neocortical pairs (membrane potential −60 to −65 mV). Thereafter increases in spike number had little additional effect on amplitude. In two of eight pairs in CA1, one involving a presynaptic basket cell and the other a putative bistratified interneuron, the fast inhibitory postsynaptic potential was blocked by bicuculline revealing a slow inhibitory postsynaptic potential that was greatly reduced by 100 μM CGP 35348 (basket cell pair). The sensitivity of this slow inhibitory postsynaptic potential to spike number was similar to that of neocortical `pure' slow inhibitory postsynaptic potentials, but was of longer duration, its plateau phase outlasting 200 ms spike trains and its maximum duration exceeding 400 ms. Computational models of GABA release, diffusion and uptake suggested that extracellular accumulation of GABA cannot alone account for the non-linear relationship between spike number and inhibitory postsynaptic potential amplitude. However, cooperativity in the kinetics of GABA B transduction mechanisms provided non-linear relations similar to experimental data. Different kinetic models were considered for how G-proteins activate K + channels, including allosteric models. For all models, the best fit to experimental data was obtained with four G-protein binding sites on the K + channels, consistent with a tetrameric structure for the K + channels associated with GABA B receptors. Thus some inhibitory connections in neocortex and hippocampus appear mediated solely by fast GABA A receptors, while others appear mediated solely by slow, non-ionotropic, possibly GABA B receptors. In addition, some inhibitory postsynaptic potentials arising in proximal portions of CA1 pyramidal cells are mediated by both GABA A and GABA B receptors. Our data indicate that the GABA released by a single interneuron can saturate the GABA B receptor mechanism(s) accessible to it and that `spillover' to extrasynaptic sites need not necessarily be proposed to explain these slow inhibitory postsynaptic potential properties.

IPSPs elicited in CA1 pyramidal cells by putative basket cells in slices of adult rat hippocampus.

CA1 basket cells are identifiable by an axonal arbour largely confined to, and spanning, the entire depth of stratum pyramidale where they innervate pyramidal somata and proximal dendrites. Basket cells display a range of electrophysiological properties and the inhibitory postsynaptic potentials (IPSPs) they elicit in pyramidal cells vary widely in duration. To determine whether these parameters are correlated, we used paired intracellular recordings, with biocytin filling, in pyramidal cells of adult hippocampal slices, and studied gamma-aminobutyric acid (GABAA) IPSPs (n = 43) elicited by putative basket cells (n = 35) with axons largely confined to stratum pyramidale in simultaneously recorded pyramidal cells. Fast-spiking interneurons elicited relatively brief IPSPs, while IPSPs elicited by burst-firing cells were amongst the slowest. Regular spiking interneurons elicited fast and slow GABAA IPSPs, but any one interneuron elicited IPSPs with remarkably similar durations in two to four pyramidal targets. However, with different types of target for a single putative basket cell, IPSPs elicited in postsynaptic interneurons were briefer than in pyramidal cells. Vertical oriens cells with somata in stratum oriens and a narrow, sparse axonal arbour in stratum pyramidale in transverse hippocampal slices, elicited IPSPs whose rise times and half widths clustered around intermediate values. Durations of IPSPs in pyramidal cells thus correlate, to a degree, with the physiological properties of presynaptic basket cells. The seven-fold range of durations observed (10-70 ms half widths) may underlie contributions made by different basket cells to hippocampal rhythms of different frequencies.