Epileptiform activity in rat hippocampal slices isolated after local septal lesioning by ibotenic acid

In surviving slices of rat hippocampus, isolated from 1 to 4 weeks after septal lesioning by ibotenic acid, extracellular and intracellular responses were recorded in region CA₃. Spontaneous and evoked epileptiform focal discharges are described, synchronous with paroxysmal depolarization shifts (PDS) of the membrane potential and with burst activity of cells. It is shown that the development of synchronized population reactions and PDS have an "all or nothing" character. The values of the resting potential and input resistance of the neurons did not differ significantly from those of cells in the control group of slices. Histological analysis showed destruction of neurons in the dorsal part of the septum, with cells of the medial septum being unaffected. The role of intraseptal mechanisms in the generation of epileptiform activity in region CA₃ of hippocampal slices is discussed.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Department of Physiology and Biochemistry, University of Pisa, Italy. Translated from Neirofiziologiya, Vol. 23, No. 5, pp. 556–564, September–October, 1991.     

Effect of anticholinergic drugs alone and in combination with nembutal on burst theta-neurons of the rabbit septum

Activity of neurones with rhythmic theta-bursts was recorded in the medial septum--diagonal band complex of the waking rabbits with intact and deafferented septum. Effects of anticholinergic (scopolamine, atropine) and cholinomimetic (physostigmine) drugs were investigated after i.v. injection. Cholinoblocking drugs in doses, suppressing the theta-rhythm in the hippocampal EEG, eliminated rhythmic activity in some cells with weak theta-modulation. Theta-bursts persisted in cells with stable continuous rhythmicity, though its regularity decreased in some of them. Strong reticular or sensory stimulation evoked an increase of burst frequency, involvement of additional septal cells into rhythmic activity and appearance of the theta-rhythm in the hippocampal EEG. Neither anticholinergic, nor cholinomimetic drugs influenced the frequency and basic characteristics of theta-bursts in any condition tested. The anticholinergic drugs have no selective effect upon low-frequency theta-bursts. The septohippocampal connections contain a significant non-cholinergic component. The theoretical concept of the septum as a sole source of the whole frequency band of the theta-rhythm is proposed.     

Unit activity of the septum and hippocampus transplanted alone or together into the anterior chamber of the rat eye

Unit activity of grafts of the septum and hippocampus, developing for 3–6 months in the anterior chamber of the eye was investigated in acute experiments on curarized orcerveau isolé rats. Whereas neurons in the transplanted septum had spontaneous activity of irregular, regular, or rhythmic bursting type, activity was absent in hippocampal grafts or consisted of very infrequent synchronized population sites. If grafts of the septum and hippocampus developed together and contact was established between them, the same types of activity developed in the hippocampus as in the septum. In many paired grafts spontaneous epileptic phenomena were observed; they were easily provoked also by electrical stimulation of one of the grafts. Superfusion with medium with a high Mg⁺⁺ concentration and low Ca⁺⁺ concentration abolished spontaneous activity in most neurons of hippocampal but not septal grafts, and also suppressed some of the epileptic phenomena, evidence of the leading role of the septum in the organization of spontaneous hippocampal unit activity.Institute of Biological Physics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 17, No. 1, pp. 61–69, January–February, 1985.     

Effect of stimulation of neocortical regions and the subiculum on hippocampal neurons in rabbits

In chronic experiments on rabbits using extracellular recording of unit activity in hippocampal area CA₁ the effects of stimulation of the subiculum, posterior cingulate cortex, and anterior and posterior nonprimary areas of the neocortex were investigated. The effects of such stimulation were compared in the intact and chronically isolated hippocampus. It is concluded from the results that direct two-way connections exist between CA₁ and the subiculum. Polysynaptic influences of the subiculum on CA₁ are realized through the entorhinal cortex, for they are not present in the isolated hippocampus. Influences of the neocortical areas studied on CA₁ are transmitted to some extent through a relay in the subiculum. The entorhinal cortex plays no part in the realization of polysynaptic effects. The effectiveness of these influences increases with removal of the principal hippocampal afferent systems.Institute of Biological Physics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 14, No. 3, pp. 315–323, May–June, 1982.     

Negative occurrence between hippocampal rhythmic slow activity and epileptiform spikes: influence of the striatum

The effects of caudate and septal stimulation on hippocampal activity were studied. Sodium penicillin was intravenously injected in order to induce a steady rate of interictal epileptic activity. Penicillin injection caused significant reduction of the rate of occurrence and duration of hippocampal rhythmic slow activity (RSA). Spontaneous RSA as well as RSA-eliciting caudate and septal stimulation induced a marked inhibition on frequency of epileptiform complexes. Lesions of the medial septal nucleus made it impossible to record RSA or to observe any sort of influence on hippocampal epileptiform activity by caudate stimulation. This result suggests that the septum is important for RSA genesis in the striato-hippocampal pathway or in the reciprocal septo-hippocampal connections.     

Role of septal and entorhinal inputs in the generation of hippocampal electrical activity in the cat sleep-wake cycle

The effects of septal lesion and entorhinal cortex section on hippocampal electrical activity during the cat sleep-wake cycle were investigated in chronic experiments. The medial portion of the septum only was found to participate in generation of this activity. Complete suppression of hippocampal theta rhythm during active wakefulness and paradoxical sleep were the main effects of septal lesion. In slow-wave sleep, the effects of septal lesion manifested in a slight attenuation of the intensity of the dominant frequency (of 1 Hz). Widespread septal lesion does not add to the changes occurring when the medial portion of the septum is so isolated. Section of the entorhinal cortex produces a sharp increase in hippocampal theta rhythm during waking and paradoxical sleep. Clearcut attenuation of delta and subdelta rhythm intensities were observed in slowwave sleep. It is postulated that under normal conditions hippocampal entorhinal input exerts a modulating effect on the genesis of hippocampal theta rhythm.I. S. Beritashvili Institute of Physiology, Academy of Sciences of the Georgian SSR, Tbilisi. Translated from Neirofiziologiya, Vol. 19, No. 5, pp. 622–630, September–October, 1987.     

Functional characteristics of the principal intrinsic systems of hippocampal connections

By extracellular recording of unit activity and electrical stimulation of unanesthetized rabbits the character of the following principal intrinsic hippocampal connections was investigated: fascia dentata with area CA₃ (series FD-CA₃) and area CA₃ with area CA₁ (series CA₃-CA₁). Differences between the functional characteristics of these two systems of connections were shown. The CA₃-CA₁ system was highly efficient, with a sharply defined boundary between subthreshold and threshold intensities of stimulation and with a wide range of active frequencies; recruiting was negligible and no sign of prolonged potentiation was present. The FD-CA₃ system was characterized by low efficiency, a wide threshold zone, and narrow range of active frequencies, slow recruiting, and long preservation of incoming influences. The probability that these functional differences depend on the morphological characteristics of the systems of connections and their possible role in the function of the hippocampus are discussed.Institute of Biological Physics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 8, No. 3, pp. 259–266, May–June, 1976.     

Comparative characteristics of septal "burst" neurons after elimination of afferent effects of the reticular formation in the rabbit

Comparative analysis of characteristics of rhythmic theta-activity in the neurones of the medial septal nucleus and nucleus of diagonal band was performed in intact rabbits after. i. v. injection of pentobarbital, and in rabbits with chronic lesion of the ascending brain-stem afferent fibers. In both conditions theta-bursts disappeared in some cells with unstable periodic rhythmic modulation; substantial population of the septal units preserved regular burst activity. Main characteristics of theta-bursts were almost identical in both states, their mean frequency decreased to 3.5 Hz. The theta-rhythm in hippocampal EEG was usually absent; but low-frequency rhythmic activity could be evoked by electrical or sensory stimulation as well as by injection of bemegrid or physostigmine. The data show that the ascending brain-stem afferents control: the frequency of the bursts in a population of septal units regarded as bursting pace-maker cells; the total number of the septal cells secondarily (synaptically) involved into rhythmic activity. The effect of pentobarbital upon theta-rhythm results from elimination of these influences upon the septal cells.     

Effect of reticular formation on hippocampal neurons (field CA1)

The reaction of field CA₁ hippocampal neurons to stimulation of the reticular formation (RF) with impulses of different frequencies was investigated in experiments on unanesthetized rabbits. The effect of electrical and sensory stimuli was compared and the effect of reticular stimulation on the sensory responses was determined. With an increase in the frequency of RF stimulation, the number of neurons of field CA₁ responding with inhibition of the activity increases. Multimodal neurons of the hippocampus depend on the reticular input to a greater degree than unimodal neurons. Neurons whose activity does not change in response to the effect of sensory stimuli also do not respond to stimulation of the RF. Neurons responding with inhibitory reactions to sensory stimulation show a higher correlation with the effects of RF stimulation than neurons with activation reactions and, especially those with "complex" responses to the effect of sensory stimuli. In a considerable number of hippocampal neurons the responses to sensory stimuli change in the course of 10–15 min after stimulation of the RF. The role of the RF in the organization of the reactions of hippocampal neurons is discussed.Division of Memory Problems, Institute of Biological Physics, Academy of Sciences of the USSR, Pushchino-on-Oke. Translated from Neirofiziologiya, Vol. 3, No. 3, pp. 227–235, May–June, 1971.     

Investigation of heterosynaptic interactions in hippocampal areas CA1, and CA3 in vitro

Heterosynaptic interactions between synapses located at a considerable distance from the cell body (perforant path) and lying close to the body of the neuron (synapses of Schaffer's collaterals and axons of the dentate fascia) on guinea pig hippocampal neurons were investigatedin vitro. It was shown by the paired stimulus method that, using stimulation of subthreshold intensity for action potential generation, spatiotemporal summation takes place in both pairs of synaptic systems. If above-threshold stimulation was used, afferents lying close to the cell body suppressed responses evoked by stimulation of distant afferents for a longer time (up to 20 msec in area CA₁ and up to 300 msec in area CA₃) than during the opposite combination of stimuli (up to 3–8 msec). After tetanization of the dentate fascia depression of responses of area CA₃ neurons to stimulation of the perforant path was observed for 2–30 min. In the remaining cases, no significant prolonged heterosynaptic posttetanic changes were observed. The possible mechanisms of these interactions are discussed.Institute of Biophysics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 11, No. 6, pp. 524–532, November–December, 1979.     

Sensory reactions of hippocampal neurons in rabbit during functional suppression of theta rythm- controlling structures

Responses to sensory stimuli were analyzed in hippocampal CA1 neurons of unanesthetized rabbits in chronic experiments before and after reversible functional blockade of the median raphe nucleus (MR) and medial septal area (MS-DB) by local microinjections of anesthetic lidocaine. The MR blockade, which resulted in an enhancement of theta-modulation of the background activity of the hippocampal neurons, was followed by a depression of sensory responsiveness (only 46.7% of the neurons reactive before the blockade retained their responses). Reactions of all types were blocked, diminished, or inverted, but inhibitory responses were affected most severely. Lidocaine injection into the MS-DB, which blocked also all brain stem afferents ascending to the hippocampus via the MS-DB input, resulted in the total absence of theta-modulation; however, responsiveness to sensory stimuli remained relatively high (76.7% of the responses were preserved); on-effects were especially resistant to the MS-DB blockade. Comparison of evoked activity in two conditions of continuous theta rhythm generation (physostigmine injection and MR blockade) revealed striking similarities. This suggests that the theta-suppressing influence of the MR (presumably serotonergic) is primarily actualized through the control of cholinergic septo-hippocampal theta-generating mechanism. The results provide support for the view that the theta-rhythm acts like an active filter in information processing performed by hippocampal neurons.     

Functional characteristics of Schaffer's collaterals studied in vivo and in vitro

A comparative analysis was made of the functional characteristics of connections between hippocampal areas CA₃ and CA₁ (Schaffer's collaterals) in experiments in vivo on unanesthetized rabbits and in vitro on surviving slices of guinea pig hippocampus, with extracellular recording in area CA₁. In the case of electrical stimulation of the collaterals in vitro, post-activational inhibition was weak, responses of inhibition of spontaneous discharges were absent, and low frequencies of stimulation were more effective than in vivo. Posttetanic changes were found more frequently in experiments in vitro and they lasted longer than in vivo. The predominant effect of tetanization under normal conditions was depression, but during incubation it was facilitation of responses. The possible causes of these differences are discussed.Institute of Biophysics, Academy of Sciences of the USSR, Puschino-on-Oka. Translated from Neirofiziologiya, Vol. 11, No. 3, pp. 208–217, May–June, 1979.     

Effect of noradrenalin and serotonin on responses of area CA3 neurons evoked in guinea pig hippocampal slices by electrical stimulation of mossy fibers

The effect of noradrenalin (NA) and serotonin (5-HT) on responses of area CA₃ cells evoked by electrical stimulation of mossy fibers was studied in slices of guinea pig hippocampus survivingin vitro. Both substances, which modify the general level and organization of spontaneous activity, also affected responses of area CA₃ cells. Changes in magnitude and structure of the response usually correlated with corresponding changes in spontaneous activity. In certain cases NA, which lowered the frequency of spontaneous activity but increased its relative content of "complex discharges" and also the number of reduced action potentials in the complex discharge, also led to an increase in the response to stimulation. 5-HT evoked periodic grouped activity in some cells and led to the appearance of such grouped discharges for the first time in the responses of other cells. Unlike NA, 5-HT caused prolonged (up to 40 min) after-facilitation of the response and an increase in spontaneous discharge frequency.Institute of Biological Physics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 14, No. 4, pp. 410–417, July–August, 1982.     

Comparison of direct influences of the perforant path on hippocampal CA1 and CA3 neurons in vitro

The efficiency of synapses of the perforant path located on terminals of apical dendrites of CA₁ and CA₃ neurons was investigated in sections of the guinea pig hippocampus in vitro. Neurons of both areas were shown to respond to stimulation of the perforant path by action potential generation. Responses of most CA₁ neurons appeared to repetitive stimulation with a frequency of up to 30–80/sec. Neurons in area CA₃ respond only to low-frequency stimulation (under 5/sec). Posttetanic potentiation of responses to stimulation of the perforant path was found in both areas of the hippocampus.Institute of Biophysics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 11, No. 4, pp. 303–310, July–August, 1979.     

Posttetanic changes in hippocampal minimal evoked potentials

Changes in the EEG induced by a single spike were recorded in the hippocampus of an unanesthetized rabbit. Summation of focal electrical activity synchronous with spontaneous single unit discharges at the symmetrical point of contralateral hemisphere revealed no stable potentials which could reflect these changes. In two cases discharges identified as activity of Shaffer's collaterals were recorded in area CA₁. Summation of post-spike changes in evoked activity recorded by the same microelectrode showed stable negative waves with an amplitute of 40–60 µV, which could have been evoked by single spikes. The curve of amplitude of the averaged evoked potentials versus near-threshold current strength stimulating the intrahippocampal pathways was not smooth in most experiments but stepwise in character. It is suggested that the minimal evoked potential corresponding to the first step (amplitude 40–80 µV) reflects a response to stimulation of one fiber. After above-threshold tetanization prolonged posttetanic potentiation of the minimal evoked potentials did not arise in CA₁ in response to stimulation of Shaffer's collaterals. Minimal evoked potentials recorded in area CA₃ in response to stimulation of the dentate fascia showed clear potentiation. The results are in agreement with the hypothesis of the synaptic localization of the mechanisms responsible for prolonged posttetanic potentiation.Brain Institute, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 9, No. 2, pp. 124–134, March–April, 1977.     

Effect of noradrenalin and serotonin on spontaneous unit activity in hippocampal area CA3 in vitro

The effect of noradrenalin and serotonin on spontaneous unit activity in hippocampal slices (area CA₃)in vitro was studied by the addition of mediators to the incubation medium. Both drugs were found to have both an inhibitory (32 and 49%, respectively) and an activating (52 and 35%, respectively) action. The effect of noradrenalin correlated with the type of spontaneous unit activity. Mainly cells without spontaneous discharges or with single irregular discharges were activated Cells with "complex discharges" were inhibited. Unlike the effect of serotonin, the activating action of noradrenalin was stimulated by the agonist phenylephrine and inhibited by the -adrenoblocker phentolamine. The action of the monoamines persisted in medium with an increased Mg⁺⁺ concentration.Institute of Biological Physics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 13, No. 5, pp. 482–490, September–October, 1981.     

Spontaneous and evoked unit activity of the septum and hippocampus transplated into the rat neocortex

Embryonic septal and hippocampal tissue was transplanted into a cavity formed by removal of part of the parietal cortex of adult rats by suction. By extracellular recording 4–6 months after the operation cells with spontaneous activity with a frequency of 3.6±0.4 Hz, characterized by an irregular, stochastic spike distribution, were detected in the graft. About 90% of cells responded to electrical stimulation of neighboring cortical areas after a latent period of 5–43 msec. The most stable responses appeared to stimulation with frequencies of 5–10 Hz; in most cases the evoked discharge was followed by a period of inhibition of spontaneous activity (100–700 msec). The same number of cells responded to tactile stimulation of the body surface and vibrissae of the recipient animal. Specific responses of different types with latent periods of between 50 and 600 msec were observed. Normalization of unit activity of intracerebral grafts compared with activity of cells in tissue developing in the anterior chamber of the eye, and their functional integration with the recipient's brain are discussed.Institute of Biological Physics, Academy of Sciences of the USSR, Pushchino-on-Oka. Translated from Neirofiziologiya, Vol. 17, No. 2, pp. 160–168, March–April, 1985.     

Spatial distribution of hippocampal response to tooth pulp stimulation and acoustic stimulation

Evoked potentials (EP) and neuronal responses produced by tooth pulp stimulation and a clicking sound were recorded at different hippocampal sites using microelectrodes in unrestrained rats. Spatial distribution of EP was found to be the same for both types of stimulation. Averaged EP consisted of a high amplitude negative preceded by a low-amplitude positive component (N₁ and P₁, respectively). Latency of the N₁ wave reached its minimum (of 27 msec) at the middle third of the molecular layer of the dentate gyrus and the outer portion of the CA₃ apical dendrites. Latency of N₁ was considerably longer in the stratum radiatum layer of the CA₁. Laminar profiles of the amplitude of the N₁ componenent of EP produced in the dentate gyrus and the CA₃ by tooth pulp stimulation resemble those observed during perforant path stimulation; in the CA₁ they are similar to those evoked by stimulating the Schaffer collaterals. Maximum amplitude of the P₁ component was observed above the pyramidal layer of the CA₁ and the hilus. Neuronal discharge pattern changed in all hippocampal regions under the effects of both tooth pulp stimulation and the clicking sound. It is deduced that information can reach the hippocampus by two routes: via a "fast" (inhibitory) pathway through the fimbria and the fornix and a slower (excitatory) path through the entorhinal cortex.P. Flexig Institute for Brain Research, Karl Marx University, Leipzig, DR. Institute of Physiology, Pecs University Medical School, Pecs, Hungary. Translated from Neirofiziologiya, Vol. 19, No. 1, pp. 36–46, January–February, 1987.     

Modulation of inhibition in the hippocampus in vivo

The nature and mechanisms of septohippocampal transmission have been elucidated by taking advantage of an in situ preparation in experiments with Sprague-Dawley rats under urethane. Both extracellular field potentials and intracellular recordings were made in CA1-3 regions of the hippocampus; and the hippocampal commissure and medial septum stimulated to evoke synaptic activity. Using muscarinic and nicotinic agonists and antagonists it was shown that both acetylcholine and medial septal activity can increase the excitability of pyramidal cells, mainly through muscarinic receptors. The effect of septal stimulation was enhanced by local application of physostigmine and reduced by intraventricular injections of hemicholinium. It was also shown that acetylcholine, when applied in the stratum pyramidale, can reduce the voltage and conductance changes observed during evoked inhibitory postsynaptic potentials (IPSP) without affecting the action of gamma-aminobutyric acid on membrane conductance and voltage. It is therefore proposed that acetylcholine can reduce evoked IPSPs through presynaptic inhibition. Evidence is also presented that medial septal stimulation can reduce the efficacy of evoked IPSPs. These observations provide further support for the existence of a cholinergic septohippocampal pathway.     

The efferent connections of the lateral septal nucleus in the guinea pig: intrinsic connectivity of the septum and projections to other telencephalic areas

Summary The distribution of efferent fibers originating in the lateral septal nucleus was investigated in guinea pigs by means of anterograde tracing with Phaseolus vulgaris-leucoagglutinin (PHA-L). Special emphasis was placed on the intraseptal fiber systems. The fibers originating from the different subnuclei of the lateral septal nucleus formed massive horizontal connections in the rostrocaudal axis. Projections to the contralateral, congruent subnuclei were also detected. In the medial septum/diagonal band of Broca complex the largest number of PHA-L-stained fibers was found after application of the tracer into the dorsal subnucleus of the lateral septal nucleus; the density of the efferent fibers decreased progressively after injection into the intermediate or ventral subnuclei. In all cases the diagonal band contained a much higher number of efferent fibers from the lateral septal nucleus than from the medial septal nucleus. In the medial septal nucleus, terminal labeling was generally sparse. Other telencephalic areas (organum vasculosum of the lamina terminalis, nucleus accumbens, bed nucleus of the stria terminalis, amygdala, hippocampal complex, and other cortical areas) contained varying numbers of labeled projections. In double-labeling experiments, a close spatial relationship between PHA-L-stained fibers and vasoactive intestinal polypeptide-immunoreactive perikarya was observed in several of these target areas.