H Golebiewski

Theta-like activity in hippocampal formation slices: cholinergic-GABAergic interaction

Brain slice preparations obtained from the rat were used to study cholinergic GABAergic interaction in mechanisms responsible for production of theta-like activity in hippocampal slices maintained in vitro. Bicuculline, a GABA-A antagonist, applied at 25 microM facilitated the effect of low concentration carbachol (25 microM) in inducing theta-like oscillations. At 100 microM, bicuculline increased the amplitude of carbachol-induced theta-like slow waves. This carbachol-bicuculline induced field potential was antagonized by a muscarinic blocker, atropine sulphate, and a GABA-A agonist, muscimol. These results provide in vitro evidence for cholinergic-GABAergic interaction in the production of hippocampal theta-like slow waves.

Theta-like activity in hippocampal formation slices : the effect of strong disinhibition of GABAA and GABAB receptors

The involvement of GABAA and GABAB receptors in neural mechanisms responsible for the production of theta rhythms in hippocampal formation (HPC) slices is addressed in the present study. In a number of papers published in the last decade, we have demonstrated that theta-like activity can be successfully recorded in the limbic cortex maintained in vitro when the cholinergic agonists, acetylcholine, carbachol or muscarine, were added to the bath. Recently, we have also shown a strong GABAA modulation of the cholinergic-induced in vitro theta-like activity. This study presents a report of the first demonstration of in vitro theta-like field responses induced a consequence of simultaneously inhibiting hippocampal GABAA and GABAB receptors. HPC slices (350 microns) were maintained in a gas-liquid interface chamber (35 degrees C). Theta-like activity was induced in the presence of bath perfusion of bicuculline (GABAA antagonist) and 2-hydroxysaclophen (GABAB antagonist). This in vitro induced field response was antagonized both by muscimol (GABAA agonist) and baclophen (GABAB agonist). In addition, the experiments presented here revealed that bicuculline/2-hydroxysaclophen-induced in vitro theta-like activity also had a strong cholinergic M1 involvement: it was abolished by hemicholinium-3 (choline transport blocker) and pirenzepine (specific antagonist of M1 receptor), but not by gallamine (specific antagonist of M2 receptor). The results of the present study provided further evidence for a strong GABAergic/cholinergic interaction in the neural mechanism responsible for production of theta-like activity in the hippocampal formation slices.

Electrical coupling underlies theta oscillations recorded in hippocampal formation slices

The role of gap junction coupling in generation of carbachol-induced theta-like activity (TLA) in hippocampal formation (HPC) slices was investigated in this study. Two gap junction (GJ) blockers, carbenoxolone (100 microM) and quinine (100 microM), were tested. Both GJ blockers abolished cholinergically induced theta-like activity and related cell discharges. Abolishing effects were observed after 40-45 min of drug perfusion. These effects were found to be slowly and partially reversible. Our results provide evidence for the contribution of gap junction communication in mechanisms of neural synchrony, underlying the production of theta oscillations in limbic cortex maintained in vitro.

The effect of carbenoxolone on hippocampal formation theta rhythm in rats: in vitro and in vivo approaches.

The role of gap junction (GJ) coupling in the generation of hippocampal formation theta rhythm was investigated in vitro, with use of brain slices, and in vivo, with use of urethane anesthetized rats. Carbenoxolone, the succinyl ester of glycyrrhetinic acid, and GJ blocker reversibly abolished hippocampal formation theta rhythm recorded in slice preparations and urethane anesthetized rats. The present study yielded novel data which demonstrated that the pattern of delay in blockage of theta rhythm after carbenoxolone treatment, and the pattern of theta recovery after administration of this agent, require a specific time period (2-3h for delay and 8-12h for recovery), one that can be demonstrated using different experimental protocols.

Window effect of temperature on carbachol-induced theta-like activity recorded in hippocampal formation in vitro

The effect of different temperatures of ACSF (18-42 degrees C) on carbachol (CCH)-induced field potentials were examined in the present study. Two hundred and thirty one experiments were performed on hippocampal formation slices maintained in a gas-liquid interface chamber. All slices were perfused with 50 microM CCH. A recording electrode was positioned in the region of CA3c pyramidal cells. The experiments gave two main findings. First, in a presence of continuous cholinergic stimulation the temperature of the bathing medium per se determined the rate of synchronization of the field potentials and pattern of EEG activity recorded. Second, within the temperature range from 33 degrees to 37 degrees C a window effect of temperature on CCH-induced theta-like activity (TLA) was noted: in this temperature range all slices tested responded only with one pattern of EEG activity-TLA. The results are discussed in light of temperature effects on hippocampal neuronal networks.

Carbachol-induced theta-like activity in entorhinal cortex slices

The present study was conducted for two purposes: the first was to evaluate whether activation of cholinergic receptors of the entorhinal cortex in vitro (complete deafferentation) with carbachol (100 microM) was capable of producing theta (theta)-like slow activity. The second purpose was to determine whether carbachol-induced slow waveforms were mediated by muscarinic or nicotinic receptors. We demonstrated that carbachol was capable of producing theta-like slow activity. This activity was not altered by nicotinic antagonists, (+)-tubocurarine and hexametonium. Atropine and scopolamine, in contrast, completely blocked in vitro induced slow waves, indicating entorhinal muscarinic receptors to be actively involved in the mechanism generating cholinergic theta rhythm.

Firing cell repertoire during carbachol‐induced theta rhythm in rat hippocampal formation slices

One hundred and seven cells were recorded extracellularly in hippocampal formation (HPC) slices during carbachol‐induced theta. The data obtained provided evidence of a population of HPC neurons which, when activated cholinergically, participate in the generation of in vitro theta. The activity patterns of in vitro recorded theta‐related cells were shown to be similar to those of theta‐related cells recorded in vivo and cells recorded in vitro during cholinergically induced theta, and non‐theta intervals were successfully classified according to previously developed criteria for in vivo recorded theta‐related cells. The current in vitro experiments showed that, in addition to theta‐on and theta‐off cells, the HPC contained cells that were probably involved in programming the appearance and duration of theta epochs and the intervals between theta epochs. These novel types of cells were termed ’gating cells’.

Electrical coupling underlies theta rhythm in freely moving cats.

The role of gap junction coupling in the generation of theta rhythms in freely moving cats was investigated in a present study. Two gap junction blockers, carbenoxolone and quinine, were administered intraperitoneally and intrahippocampally; both gap junction blockers abolished or diminished (respectively) hippocampal formation theta. The inhibitory effect developed approximately 30 min after drug administration. This effect was found to be reversible. Our results provide the first direct in vivo evidence for the contribution of gap junction communication in mechanisms of neural synchrony, underlying the production of theta in in vivo conditions.

Muscarinic (M1) mediation of hippocampal spontaneous theta rhythm in freely moving cats

The effects of intrahippocampally applied different doses of muscarinic (atropine sulphate, pirenzepine, gallamine) and nicotinic (hexamethonium, mecamylamine) antagonists on the spontaneous theta rhythm in the cat hippocampal formation were investigated. The injections of atropine and pirenzepine abolished spontaneous theta but administration of gallamine did not affect the EEG pattern. The intrahippocampally administered nicotinic blockers, hexamethonium and mecamylamine, were completely ineffective in antagonizing theta waves. The data suggest that the spontaneous theta rhythm in the cat is mediated by the M1 receptor subtype.

Is the dentate gyrus an independent generator of in vitro recorded theta rhythm

In this study we investigated the ability of the dentate gyrus to independently generate cholinergically induced theta rhythm in vitro. Two different experiments were performed. In Experiment I, new laminar profiles of theta phase, amplitude, and current sources and sinks were constructed. In this experiment a gradual phase shift of theta waves in the CA1 stratum radiatum was observed. Simultaneously, two amplitude maxima were detected: the first in the CA1 stratum oriens, and the second in the CA1 stratum lacunosum-moleculare. Moreover, during the positive peak of theta in the CA1 stratum oriens, two large sinks were observed: the first localized in the stratum oriens and the second in the stratum lacunosum-moleculare. In Experiment II the EEG activity of three different transected hippocampal slices (CA1 transected slices, CA3c transected slices, and DG transected slices) was recorded. It was demonstrated that the dentate gyrus granular cell body layer was not able to independently produce in vitro theta rhythm. Data obtained in both experiments provide strong evidence that in cholinegically treated hippocampal formation maintained in vitro there is no independent generator of theta rhythm in the region of the dentate gyrus granular cell layer.