Lai-Wo Stan Leung

Intrinsic membrane potential oscillations in hippocampal neurons in vitro

Membrane potential oscillations (MPOs) of 2-10 Hz and up to 6 mV were found in almost all stable hippocampal CA1 and CA3 neurons in the in vitro slice preparation. MPOs were prominent for pyramidal cells but less pronounced in putative interneurons. MPOs were activated at threshold depolarizations that evoked a spike and the frequency of the MPOs increased with the level of depolarization. MPOs were distinct from and seemed to regulate spiking, with a spike often riding near the top of a depolarizing MPO wave. Analysis of the periodicity of the oscillations indicate that the period of MPOs did not depend on the afterhyperpolarization (AHP) following a single spike. MPOs persisted in low (0-0.1 mM) Ca2+ medium, with or without Cd2+ (0.2 mM), when synaptic transmission was blocked. Choline-substituted low-Na+ (0-26 mM) medium, 3 microM tetrodotoxin (TTX) or intracellular injection of QX-314 reduced or abolished the fast Na(+)-spike and reduced inward anomalous rectification. About 40% of CA1 neurons had no MPOs after Na+ currents were blocked, suggesting that these MPOs were Na(+)-dependent. In about 60% of the cells, a large depolarization activated Ca(2+)-dependent MPOs and slow spikes. MPOs were not critically affected by extracellular Ba2+ or Cs2+, or by 0.2 mM 4-aminopyridine, with or without 2 mM tetraethylammonium (TEA). However, in 5-10 mM TEA medium, MPOs were mostly replaced by 0.2-3 Hz spontaneous bursts of wide-duration spikes followed by large AHPs. Low Ca2+, Cd2+ medium greatly reduced the spike width but not the spike-bursts. In conclusion, each cycle of an MPO in normal medium probably consists of a depolarization phase mediated by Na+ currents, possibly mixed with Ca2+ currents activated at a higher depolarization. The repolarization/hyperpolarization phase may be mediated by Na+/Ca2+ current inactivation and partly by TEA-sensitive, possibly the delayed rectifier, K+ currents. The presence of prominent intrinsic, low-threshold MPOs in all hippocampal pyramidal neurons suggests that MPOs may play an important role in information processing in the hippocampus.

Electrical activity of the cingulate cortex. I. Generating mechanisms and relations to behavior

Spontaneous slow waves (EEG) and multiple unit activity (MUA) were recorded in the posterior cingulate cortex (area 29) and the dorsal hippocampus of the freely moving rat by means of chronically implanted electrodes. Three different wave patterns were discerned in the cingulate EEG. Irregular slow waves occurred during grooming, drinking, eating (Type II behavior) and slow-wave sleep (SWS). The irregular waves also contained sharp transients of about 20 ms duration called EEG-spikes. EEG-spikes reversed their polarity within the cingulate cortex and correlated with an increase in cingulate MUA. They were probably generated by deep (layer IV to VI) neurons in the cingulate cortex. Theta rhythm of 6-10 Hz accompanied walking, rearing, postural shifts, head movements (Type I behavior) and rapid-eye-movement sleep (REMS). MUA of low-amplitude units was phase-locked to the local theta waves, suggesting local generation of the slow waves. However, volume-conduction from the hippocampus would likely contribute to the cingulate theta since no reversal of the theta waves was found in the cingulate cortex. Fast waves of greater than 30 Hz were generally larger during Type I than during Type II behavior. Cellular generators for fast waves are not known. High-amplitude (greater than 100 microV) MUA only appeared during Type II behavior, and in particular during SWS. During REMS, these units were silent. Stimulation of the contralateral homotopic cingulate cortex gave antidromic and synaptic components in the average evoked potential (AEP). The long latency waves of the AEP varied with behaviors and appeared oscillatory (25-40 Hz) during Type I but not during Type II behavior. In summary, the cingulate cortex has a rich gamut of spontaneous and evoked electrical activities which bears some resemblance to that of the hippocampus.

APV, an N-methyl-D-aspartate receptor antagonist, blocks the hippocampal theta rhythm in behaving rats.

2-Aminophosphonovaleric acid (APV), an N-methyl-D-aspartate (NMDA) receptor antagonist, was infused into the lateral ventricles of behaving rats. A 10 or 20 microgram dose of APV attenuated the hippocampal theta rhythm and the theta phase-shift at the apical dendrites of hippocampal CA1 region. A selective suppression of the atropine-sensitive theta rhythm was suggested.

Factors affecting paired-pulse facilitation in hippocampal CA1 neurons in vitro

Factors underlying paired-pulse facilitation (PPF) were studied by intracellular and field recordings of CA1 neurons in the hippocampal slice in vitro, following stimulation of the Schaffer collaterals apical dendritic afferents. Similar magnitudes of PPF were found using the slopes or peaks of the excitatory postsynaptic potentials (EPSPs) recorded intracellularly or extracellularly at the soma or dendrites. The paired-pulse EPSP facilitation index (EPI), defined as the ratio of EPSP slope evoked by the second pulse (E2) to that evoked by the first pulse (E1), had a broad peak at 30-60 ms interpulse interval (IPI). EPI was largest at small E1 and decreased with an E1 increase. Spiking excitability was enhanced after the second as compared to the first pulse as evidenced by (1) a decreased latency to fire and (2) an increased tendency to fire double or multiple spikes. The PPF of spiking resulted partly from an increased E2 and partly from a diminished inhibition evoked by the second pulse. Whether the first pulse elicited a spike or not had no significant effect on the EPI. Multiple spiking evoked by the second pulse was partly blocked by the GABAB antagonist CGP35348 (1 mM). The PPF of the EPSP slopes, however, was not significantly affected by GABAB antagonists, GABAA antagonist bicuculline or NMDA antagonist 2-aminophosphonovalerate. In conclusion, PPF may serve as a means of amplification of synaptic transmission such that reliable spike output may result from a given set of synapses.

The role of serotonin in the control of cerebral activity: studies with intracerebral 5,7-dihydroxytryptamine

Intact rats treated with centrally acting antimuscarinic (atropinic) drugs display large amplitude irregular slow waves in both the neocortex and hippocampus during behavioral immobility and some stereotyped automatic behaviors (Type 2 behavior). However, rhythmical slow activity in the hippocampus and low voltage fast activity in the neocortex occur in close correlation with spontaneous changes in posture, head movement, walking, rearing, swimming or struggling when held (Type 1 behavior). It has previously been proposed that these waveforms, jointly referred to as atropine-resistant cerebral activation (ARCA) are dependent on ascending serotonergic projections. As a further test of this hypothesis, we have studied rats in which forebrain levels of serotonin and 5-hydroxyindoleacetic acid were reduced to 3-10% of control levels as a result of multiple intrabrainstem injections of 5,7-dihydroxytryptamine. This treatment strongly reduced or abolished ARCA in most cases but did not reduce atropine-sensitive cerebral activation which appears to be dependent on ascending cholinergic projections from the basal forebrain to the cerebral cortex. Therefore, ARCA appears to be dependent on ascending serotonergic inputs to the forebrain.

Pathways through cingulate, neo- and entorhinal cortices mediate atropine-resistant hippocampal rhythmical slow activity.

Rats prepared with a lesion separating the entorhinal cortex from the neocortex and cingulate cortex displayed apparently normal hippocampal rhythmical slow activity (RSA) with a frequency of 6-12 Hz in both CA1 and dentate gyrus during Type 1 behavior (locomotion, head movements, changes in posture). Variations in the commissural average evoked potential (AEP) and increased power in the 30-100 Hz range (fast waves) also correlated with Type 1 behavior. Urethane did not abolish the RSA. However, systemic administration of atropinic drugs eliminated all RSA and eliminated or attenuated the Type 1 behavior-related variations in the AEP and fast waves. Thus, the normally present atropine-resistant RSA was eliminated by the cortical lesion while atropine-sensitive RSA remained intact. Removal of cingulate cortex alone was partially effective in suppressing atropine-resistant RSA but a lesion of the neocortex only, sparing cingulate cortex, had a minimal effect on it. Lesions of the amygdala, the anterior or medial thalamus or the cerebellum had little or no effect on atropine-resistant RSA. Previous work has shown that lesions of the entorhinal cortex or lateral hypothalamus eliminate atropine-resistant RSA. We suggest that atropine-resistant RSA is mediated by a somewhat diffuse pathway which traverses the hypothalamus, cingulate cortex, and neocortex before reaching the hippocampus via the entorhinal cortex.

Electrical activity of the cingulate cortex. II. Cholinergic modulation

The role of the cholinergic innervation in the modulation of cingulate electrical activity was studied by means of pharmacological manipulations and brain lesions. In the normal rat, an irregular slow activity (ISA) accompanied with EEG-spikes was recorded in the cingulate cortex during immobility as compared to walking. Atropine sulfate, but not atropine methyl nitrate, increased ISA and the frequency of cingulate EEG-spikes. Pilocarpine suppressed ISA and EEG-spikes during immobility, and induced a slow (4-7 Hz) theta rhythm. Unilateral or bilateral lesions of the substantia innominata and ventral globus pallidus area using kainic acid did not significantly change the cingulate EEG or its relation to behavior. Large electrolytic lesions of the medial septal nuclei and vertical limbs of the diagonal band generally decreased or abolished all theta activity in the cingulate cortex and the hippocampus. However, in 5 rats the cingulate theta rhythm increased while the hippocampal theta disappeared after a medial septal lesion. The large, postlesion cingulate theta, accompanied by sharp EEG-spikes during its negative phase, is an unequivocal demonstration of the existence of a theta rhythm in the cingulate cortex, independent of the hippocampal rhythm. Cholinergic afferents from the medial septum and diagonal band nuclei are inferred to be responsible for the behavioral suppression of cingulate EEG-spikes and ISA, and partially for the generation of a local cingulate theta rhythm. However, an atropine-resistant pathway and a theta-suppressing pathway, possibly coming from the medial septum or the hippocampus, may also be important in cingulate theta generation.

Hippocampal electrical activity following local tetanization. I. Afterdischarges

Following a short (1-10 s) train of repetitive stimulation delivered to the hippocampal CA1 region, the following sequelae of afterdischarges (ADs) was seen: (1) a silent period of 2-4 s, (2) a large primary (1 degree) AD usually alvear-surface negative and deep positive, (3) a period of suppressed hippocampal EEG, (4) a secondary (2 degrees) hippocampal AD, and after 3-6 min, (5) 15-25 min of enhanced (up to 10 times normal) fast (30-70 Hz) waves. The 2 degrees hippocampal AD was preceded by or simultaneous with large AD at the amygdaloid electrodes. Electrolytic lesions (n = 7) or large heat lesions of the amygdala (n = 5) or electrolytic lesions of the medial septum (n = 10) were not successful in suppressing the 2 degrees hippocampal AD. However, 4 rats with radiofrequency lesion and 3 rats with bilateral aspiration lesion of the entorhinal cortex had diminished or no 2 degrees hippocampal AD. The fast waves after tetanization were reversed 180 degrees across surface and deep CA1 electrodes. The fast wave increase was blocked by atropine sulfate (25-50 mg/kg i.p.), scopolamine hydrochloride (5 mg/kg i.p.) and medial septal lesions. It was concluded that the 2 degrees hippocampal AD may depend on a reverberation of neural circuitry involving the entorhinal cortex. The 2 degrees AD recorded from amygdala electrodes may partly reflect spreading of activities from the entorhinal cortex. On the other hand, the increase in fast waves after tetanization requires an intact septohippocampal, muscarinic cholinergic input, and may depend on an enhanced cholinergic input or an increased response.

Involvement of the nucleus accumbens-ventral pallidal pathway in postictal behavior induced by a hippocampal afterdischarge in rats

The hypothesis that postictal motor behaviors induced by a hippocampal afterdischarge (AD) are mediated by a pathway through the nucleus accumbens (NAC) and ventral pallidum (VP) was evaluated in freely moving rats. Tetanic stimulation of the hippocampal CA1 evoked an AD of 15-30 s and an increase in number of wet-dog shakes, face washes, rearings and locomotor activity. Bilateral injection of haloperidol (5 micrograms/side) or the selective dopamine D2 receptor antagonist, (+/-)-sulpiride (200 ng/side) before the hippocampal AD, into the NAC selectively reduced rearings and locomotor activity, but not the number of wet-dog shakes and face washes. Injection of R(+)-SCH-23390 (1 microgram/side), a D1 receptor antagonist, or rimcazole (0.4 mg/side), a sigma opioid receptor antagonist, into the NAC did not significantly alter postictal behaviors. Bilateral injection of muscimol (1 ng/side), a gamma-aminobutyric acid (GABAA) receptor agonist, into the VP before the AD significantly blocked all postictal behaviors. It is concluded that postictal locomotor activity induced by a hippocampal AD is mediated by activation of dopamine D2 receptors in the NAC and a pathway through the VP.

Spontaneous hippocampal interictal spikes following local kindling: time-course of change and relation to behavioral seizures

Spontaneous interictal spikes (SISs) were recorded in the hippocampus in freely behaving rats following hippocampal stimulations that resulted in afterdischarges (ADs). Hippocampal SISs were detected after an average of 5 (range 2-10) daily ADs. The rate of SISs typically increased minutes after a tetanus, and then decayed with time constants of approximately 70 min and 1.5 days. Seizure onset in the kindling paradigm was not related to a consistent change in SIS rate. Following the interruption of daily kindling, SIS rate invariably decreased to near zero by 4-8 days while seizure susceptibility, as tested by the ability to evoke generalized convulsions, remained unchanged. Despite having a low or zero SIS rate the hippocampus seemed to retain an excitability after kindling interruption, as demonstrated by the observation that an average of 1.7 rekindling stimulations resulted in a high SIS rate. In conclusion, changes in hippocampal SISs were closely time-locked to an AD, and not to evoked behavioral seizures. Hippocampal SISs probably reflect an excitability change that is more local than that necessary for evoking behavioral convulsions. The persistence of SISs in terms of hours and days suggests the involvement of long-term potentiation.