Presynaptic kainate receptors that enhance the release of GABA on CA1 hippocampal interneurons.

We report that kainate receptors are present on presynaptic GABAergic terminals contacting interneurons and that their activation increases GABA release. Application of kainate increased the frequency of miniature inhibitory postsynaptic currents recorded in CA1 interneurons. Local applications of glutamate but not of AMPA or NMDA also increased GABA quantal release. Application of kainate as well as synaptically released glutamate reduced the number of failures of GABAergic neurotransmission between interneurons. Thus, activation of presynaptic kainate receptors increases the probability of GABA release at interneuron-interneuron synapses. Glutamate may selectively control the communication between interneurons by increasing their mutual inhibition.

GluR5 kainate receptor activation in interneurons increases tonic inhibition of pyramidal cells.

We studied the modulation of GABAergic inhibition by glutamate and kainate acting on GluR5-containing kainate receptors in the CA1 hippocampal region. Glutamate, kainate or ATPA, a selective agonist of GluR5-containing receptors, generates an inward current in inhibitory interneurons and cause repetitive action potential firing. This results in a massive increase of tonic GABAergic inhibition in the somata and apical dendrites of pyramidal neurons. These effects are prevented by the GluR5 antagonist LY 293558. Electrical stimulation of excitatory afferents generates kainate receptor-mediated excitatory postsynaptic currents (EPSCs) and action potentials in identified interneurons that project to the dendrites and somata of pyramidal neurons. Therefore glutamate acting on kainate receptors containing the GluR5 subunit may provide a protective mechanism against hyperexcitability.

Dendritic but not somatic GABAergic inhibition is decreased in experimental epilepsy.

Impaired inhibition is thought to be important in temporal lobe epilepsy (TLE), the most common form of epilepsy in adult patients. We report that, in experimental TLE, spontaneous GABAergic inhibition was increased in the soma but reduced in the dendrites of pyramidal neurons. The former resulted from the hyperactivity of somatic projecting interneurons, whereas the latter was probably due to the degeneration of a subpopulation of dendritic projecting interneurons. A deficit in dendritic inhibition could reduce seizure threshold, whereas enhanced somatic inhibition would prevent the continuous occurrence of epileptiform activity.

Maturation of firing pattern in chick vestibular nucleus neurons.

The principal cells of the chick tangential nucleus are vestibular nucleus neurons participating in the vestibuloocular and vestibulocollic reflexes. In birds and mammals, spontaneous and stimulus-evoked firing of action potentials is essential for vestibular nucleus neurons to generate mature vestibular reflex activity. The emergence of spike-firing pattern and the underlying ion channels were studied in morphologically-identified principal cells using whole-cell patch-clamp recordings from brain slices of late-term embryos (embryonic day 16) and hatchling chickens (hatching day 1 and hatching day 5). Spontaneous spike activity emerged around the perinatal period, since at embryonic day 16 none of the principal cells generated spontaneous action potentials. However, at hatching day 1, 50% of the cells fired spontaneously (range, 3 to 32 spikes/s), which depended on synaptic transmission in most cells. By hatching day 5, 80% of the principal cells could fire action potentials spontaneously (range, 5 to 80 spikes/s), and this activity was independent of synaptic transmission and showed faster kinetics than at hatching day 1. Repetitive firing in response to depolarizing pulses appeared in the principal cells starting around embryonic day 16, when <20% of the neurons fired repetitively. However, almost 90% of the principal cells exhibited repetitive firing on depolarization at hatching day 1, and 100% by hatching day 5. From embryonic day 16 to hatching day 5, the gain for evoked spike firing increased almost 10-fold. At hatching day 5, a persistent sodium channel was essential for the generation of spontaneous spike activity, while a small conductance, calcium-dependent potassium current modulated both the spontaneous and evoked spike firing activity. Altogether, these in vitro studies showed that during the perinatal period, the principal cells switched from displaying no spontaneous spike activity at resting membrane potential and generating one spike on depolarization to the tonic firing of spontaneous and evoked action potentials.

Newly formed excitatory pathways provide a substrate for hyperexcitability in experimental temporal lobe epilepsy.

Temporal lobe epilepsy (TLE) in humans and animals is associated with axonal sprouting of glutamatergic neurons and neosynaptogenesis in the hippocampal formation. We examined whether this plasticity of excitatory pathways contributes to an increased level of glutamatergic excitation in the CA1 region of rats experiencing chronic spontaneous limbic seizures following kainic acid or pilocarpine treatment. In chronic cases, we report an extensive axonal sprouting of CA1 pyramidal neurons, with many axonal branches entering the pyramidal cell layer and stratum radiatum, regions that are not innervated by axonal collaterals of CA1 pyramidal neurons in control animals. Concurrently with this anatomical reorganization, a large increase of the spontaneous glutamatergic drive is observed in the dendrites and somata of CA1 pyramidal cells. Furthermore, electrical activation of the reorganized CA1 associational pathway evokes epileptiform bursts in CA1 pyramidal cells. These findings suggest that reactive plasticity could contribute to the hyperexcitability of CA1 pyramidal neurons and to the propagation of seizures in these two models of TLE.

A synaptically evoked late hyperpolarization in the rat dorsolateral geniculate neurons in vitro.

Intracellular potentials were recorded from presumed relay neurons in the rat dorsolateral geniculate nucleus maintained in vitro preparations. In this material, the neuronal circuit includes the excitatory optic tract which innervates monosynaptically both relay and intrinsic neurons, the latter providing a feed-forward GABAergic inhibition on the former. Electrical stimulation of the optic tract evokes in the dorsolateral geniculate neurons an early excitatory postsynaptic potential followed by an inhibitory postsynaptic potential which precedes a so far unreported long-lasting late hyperpolarization. The properties of the inhibitory postsynaptic potential are consistent with the notion that they are of disynaptic (feed-forward) origin and that they are the consequence of GABAA receptor activation. In contrast, the late hyperpolarization, which was found in almost every neuron, was enhanced by GABAA blockers, without accompanying changes in the resting membrane potential or the input resistance of the recorded cells. The late hyperpolarization had a lower threshold than the excitatory postsynaptic potential, a long latency (m = 38 +/- 4 ms, n = 10) and was of long duration (m = 308 +/- 57 ms, n = 10). The occurrence and threshold for producing these two potentials were uncorrelated, and paired stimulations of the optic tract showed a marked difference of their recovery time-courses. The late hyperpolarization could be elicited only by afferent stimulations; it never followed intracellularly induced depolarizations and/or anodal break calcium spikes. It was associated with a small conductance increase, sufficient, however, to inhibit high-frequency discharges induced by intracellular injection of depolarizing currents. The late hyperpolarization decreased in amplitude with membrane hyperpolarization and ultimately reversed polarity. The apparent reversal potential followed shifts in extracellular potassium concentration in an almost Nernstian relation (47 mV for a tenfold increase in [K]0). Involvement of GABAB receptors in the generation of this potential may be postulated since baclofen readily hyperpolarized the neurons and decreased their input resistance in the presence of GABAA blockers. We conclude that the late hyperpolarization is a postsynaptic potential mediated by an increased conductance to K ions. Our results further suggest that a minimal disynaptic feed-forward circuit impinging on the relay neurons of the dorsolateral geniculate nucleus is sufficient to subserve this late hyperpolarization.(ABSTRACT TRUNCATED AT 400 WORDS)

Oscillations of the spontaneous slow-wave sleep rhythm in lateral geniculate nucleus relay neurons of behaving cats.

Intracellular recordings of 31 lateral geniculate nucleus relay neurons were performed in darkness in behaving cats in order to analyse electrical postsynaptic events which appeared during slow-wave sleep. A specific pattern characterized slow-wave sleep: a rapid depolarizing potential arising from baseline initiated a slow depolarization lasting for 40-60 ms which in turn most often elicited delayed fast spikes. This pattern recurred at a frequency of 6-12/s. The slow depolarizations were voltage dependent, usually not separated by any obvious phasic hyperpolarization and showed refractoriness. Other rapid depolarizing potentials occurring during the time course or at the end of a slow depolarization could have generated spike(s) but were followed by a rapid decay. Slow depolarizations were not observed during arousal or paradoxical sleep when the neurons tonically depolarized and displayed either rapid depolarizing potentials with a fast decay or repetitive firing and long high frequency bursts. In five of the studied neurons, decreases in frequency of the spontaneous rapid depolarizing potentials occurred during slow-wave sleep for 3-30 s oscillatory periods without any change in the behavioural state. During these periods all of the few remaining rapid depolarizing potentials arose from a flat baseline, had a higher amplitude and initiated a slow depolarization which always elicited a spike or burst of spikes after a brief delay. The slow-wave sleep rhythm decreased to 1-5/s. Simultaneously the baseline membrane potential hyperpolarized by a few millivolts and reached a level for reversal of inhibitory postsynaptic potentials. Imposed hyperpolarization of the membrane during wakefulness did not reveal any slow depolarization. But strong synaptic excitatory inputs and direct excitation (a break of the current pulse) from a hyperpolarized membrane did evoke the slow depolarization and eventually the fast spike(s) in both control and oscillatory neurons. A rhythm similar to that of slow-wave sleep was elicited during wakefulness by optic tract stimulation and was enhanced by membrane hyperpolarization. But under these conditions the rhythm was initiated by a phasic hyperpolarization and was composed of an alternating hyperpolarization-depolarization. Spontaneously and synaptically evoked rapid depolarizing potentials arising from baseline had a similar rising slope. The spontaneous ones initiated a slow depolarization leading to fast spike(s) during slow-wave sleep and could directly generate fast spike(s) during wakefulness.(ABSTRACT TRUNCATED AT 400 WORDS)

Modulation of postsynaptic activities of thalamic lateral geniculate neurons by spontaneous changes in number of retinal inputs in chronic cats. 1. Input-output relations.

The experiments were designed to explore the role of retinal inputs compared with that of the behavioral state in the modulation of the output of thalamic lateral geniculate neurons during sleep and wakefulness in cats with intact visual pathways. We made the following assumptions: the retinal dark discharge, while showing spontaneous pauses in activity, does not vary with the behavioral state; the optic tract inputs postsynaptically elicit subthreshold activities called S-potentials which in turn generate spikes, the degree of transformation being dependent on the level of alertness. On the basis of these assumptions, it could be expected that changes in retinal input frequency would modify the rate of the S-potentials. Therefore the effect of spontaneous decreases in frequency of S-potentials on the spike rate and pattern was examined in juxta- and intracellular recordings from chronically implanted cats during natural sleep and wakefulness. During quiet wakefulness and light slow-wave sleep, lateral geniculate relay neurons normally displayed numerous S-potentials associated with a moderate firing rate. Many neurons occasionally showed transient reductions in frequency of the S-potentials and an oversimplification of the discharges which combined a decreased rate with a prevalent rhythmical burst pattern. Antidromic responsiveness remained unchanged. The oscillatory periods recurred two to six times without any alteration in the control state level. They were not observed throughout wakefulness and paradoxical sleep, during which neuronal activity combined a high spike rate with a low S-potential rate. The modifications were confirmed by computation of the mean rates and of the inter-event intervals. The transfer ratio (spikes/S-potentials + spikes) significantly increased both during the oscillatory periods poor in S-potentials of quiet wakefulness and during active wakefulness. But the correlation between the transfer ratio and the spike frequency, which was high throughout the control behavioral states, faded during the periods poor in S-potentials. Thus the transient falls in the frequency of S-potentials which occurred spontaneously during quiet wakefulness caused burst discharges in lateral geniculate relay neurons, which resembled a sleep deepening, but also paralleled the effect of experimental deafferentation. The data indicate that the iterative spikes grouped in well spaced bursts which persisted during decreases in subthreshold postsynaptic activities result in an enhanced signal-to-noise ratio.(ABSTRACT TRUNCATED AT 400 WORDS)

Spontaneous synaptic activity in chick vestibular nucleus neurons during the perinatal period.

The principal cells of the chick tangential nucleus are second-order vestibular neurons involved in the vestibuloocular and vestibulocollic reflexes. The spontaneous synaptic activity of morphologically identified principal cells was characterized in brain slices from 1-day-old hatchlings (H1) using whole-cell voltage-clamp recordings and Cs-gluconate pipet solution. The frequency was 1.45 Hz for spontaneous excitatory postsynaptic currents (sEPSCs) and 1.47 Hz for spontaneous inhibitory postsynaptic currents (sIPSCs). Using specific neurotransmitter receptor antagonists, all of the sEPSCs were identified as AMPA receptor-mediated events, whereas 56% of the sIPSCs were glycine and 44% were GABA(A) receptor-mediated events. On exposure to TTX, the frequency of EPSCs decreased by 68%, while the frequency of IPSCs decreased by 33%, indicating greater EPSC dependency on presynaptic action potentials. These data on spontaneous synaptic activity at H1 were compared with those obtained in previous studies of 16-day old embryos (E16). After birth, the spontaneous synaptic activity exhibited increased EPSC frequency, increased ratio for excitatory to inhibitory events, increased percentage of TTX-dependent EPSCs, and faster kinetics. In addition, the ratio for glycine/GABA receptor-mediated events increased significantly. Altogether, these data indicate that at hatching spontaneous synaptic activity of vestibular nucleus neurons in brain slices of the chick tangential nucleus undergoes appreciable changes, with increased frequency of EPSCs and glycinergic activity playing more important roles compared with the late-term chick embryo when GABAergic activity prevailed. The definition of this developmental pattern of synaptic activity in vestibular nucleus neurons should contribute to understanding how vestibular reflex activity is established in the hatchling chick.

Distribution of spontaneous currents along the somato-dendritic axis of rat hippocampal CA1 pyramidal neurons.

Excitatory and inhibitory pathways have specific patterns of innervation along the somato-dendritic axis of neurons. We have investigated whether this morphological diversity was associated with variations in the frequencies of spontaneous and miniature GABAergic and glutamatergic synaptic currents along the somato-dendritic axis of rat hippocampal CA1 pyramidal neurons. Using in vitro whole cell recordings from somata, apical dendrites and basal dendrites (for which we provide the first recordings) of CA1 pyramidal neurons, we report that over 90% of the spontaneous currents were GABAergic, <10% being glutamatergic. The frequency of spontaneous GABAergic currents was comparable in the soma and in the dendrites. In both somata and dendrites, the Na(+) channel blocker tetrodotoxin abolished more than 80% of the spontaneous glutamatergic currents. In contrast, tetrodotoxin abolished most dendritic (>90%) but not somatic (<40%) spontaneous GABAergic currents. Computer simulations suggest that in our experimental conditions, events below 40pA are electrotonically filtered to such a degree that they are lost in the recording noise. We conclude that, in vitro, inhibition is massively predominant over excitation and quantitatively evenly distributed throughout the cell. However, inhibition appears to be mainly activity-dependent in the dendrites whereas it can occur in the absence of interneuron firing in the soma. These results can be used as a benchmark to compare values obtained in pathological tissue, such as epilepsies, where changes in the balance between excitation and inhibition would dramatically alter cell behaviour.