Hippocampal rhythmical slow activity following ibotenic acid lesions of the septal region. I. Relations to behavior and effects of atropine and urethane

The effects of intraseptal injections of various concentrations of ibotenic acid on hippocampal electrical activity were studied in freely moving and urethane-anesthetized rats. Ibotenic acid selectively abolished the atropine-sensitive form of hippocampal rhythmical slow activity (RSA) normally seen during urethane anesthesia. Large amplitude irregular activity (LIA) and RSA in the waking state were somewhat depressed as well. Despite this, clear RSA persisted in the waking state in association with locomotion or struggling (Type 1 behavior). As in normal rats, such RSA was resistant to systemic administration of atropine. Analysis of brain sections stained with gallocyanin or for acetylcholinesterase showed that ibotenic acid produced cell loss in the dorsal lateral septal nucleus and the septohippocampal nucleus. Cells in the medial septal and diagonal band nuclei were resistant to ibotenic acid. The results suggest that intrinsic septal circuitry is critically involved in the generation of the atropine-sensitive (presumably cholinergic) form of RSA. The mechanisms by which LIA and the two forms of RSA are generated in the hippocampus is discussed.     

The effects general and restricted serotonergic lesions on hippocampal electrophysiology and behavior

Depletion of the forebrain serotonergic system was found in previous studies to induced an increased excitability of the dentate gyrus (DG) granule cells and, when combined with a cholinergic deficiency, to impair spatial learning. We now compared the effects of general forebrain serotonergic lesions induced by intracerebroventricular injection of 5,7-dihydroxytryptamine (5,7-DHT), to those of a more restricted injection of 5,7-DHT into fornix-fimbria and cingulum, to eliminate hippocampal serotonergic innervation. Control and lesioned rats were injected with atropine and tested in the spatial learning water-maze task. Following the behavioral tests, rats were anesthetized and the responsiveness of the DG to perforant path (PP) stimulation was measured. To assess the lesions functionally, responses to application of the serotonin releasing drug fenfluramine (FFA) were measured. Finally, the reduction, in the hippocampus of serotonergic innervation was evaluated by [³H]imipramine binding. The effects of the lesions on the responsiveness to FFA confirmed that the ICV lesions were functionally more general than the FF lesions. [³H]Imipramine binding indicated that both lesions reduced the sertonergic innervation of the hippocampus significantly. Behaviorally, both lesioned groups were impaired in the water-maze. Electrophysiologically, in both DG excitability was higher than in controls and in both hyperexcitability was associated with an increase in feed-forward inhibition. The results suggest that the serotonergic innervation of the hippocampus proper is involved in cognitive functions associated with the hippocampus.     

Reset of hippocampal rhythmical activities by afferent stimulation

Physostigmine induced theta rhythm and unit activity were recorded from the dorsal hippocampus in immobilized locally anesthetized rats. Correlations between theta and rhythmical unit activity and their modifications by hippocampal afferent stimulation were studied. The principal finding was that electrical stimulation of afferents reset theta and rhythmical unit activity in phase. Poststimulus theta displayed a variable frequency which depended upon the structure stimulated. Lower frequencies were evoked by septal, higher frequencies by entorhinal and reticular formation stimulations. When theta rhythms were absent either by spontaneous disappearance or as a consequence of lesions in the fornix superior of septum, the reset was not observed. The reset of the theta rhythms and unit activity by afferent stimulation, suggests that the hippocampus may participate in timing mechanisms.     

Effects of atropine on hippocampal theta cells and complex-spike cells

The medial septal nuclei are essential for the naturally occurring hippocampal theta rhythm. Evidence that the rhythmic activity of the septum is carried via cholinergic afferents to the hippocampus has been: (a) the existence of a cholinergic septo-hippocampal projection, and (b) the sensitivity of one type of theta rhythm to antimuscarinic agents or cholinergic depletion. The muscarinic action of acetylcholine on pyramidal cells, however, is too slow to carry even a 4 Hz signal. Recent in vitro studies have confirmed a fast excitatory response by some hippocampal interneurons to muscarinic agonists. In urethane anesthetized rats, iontophoretic application of atropine to 17 hippocampal theta cells (presumed interneurons) during the theta rhythm, reduced their firing rates to an average of 24% of control rates. The effect of iontophoretic atropine application to 4 CA1 complex-spike cells (presumed pyramidal cells) was a selective elimination of their bursting activity with no significant effect on overall firing rate. The data suggest that: (1) interneuronal firing, during the hippocampal theta rhythm, is dominated by an excitatory cholinergic input and not by excitatory collaterals of pyramidal cells; and (2) somatic burst firing by CA1 pyramidal cells requires the presence of acetylcholine.     

Colchicine-induced deafferentation of the hippocampus selectively disrupts cholinergic rhythmical slow wave activity

It has been proposed that hippocampal rhythmical slow wave activity (RSA or theta-rhythm) induced by sensory stimulation (atropine-sensitive theta) is generated by the cholinergic septo-hippocampal system. Although ablations of the septum or its projections to the hippocampus disrupt hippocampal RSA, such non-selective lesions damage both cholinergic and non-cholinergic septo-hippocampal inputs. The present study assesses the effects of a selective septal neurotoxic lesion on hippocampal electrical activity. Colchicine, which has been reported to be selectively toxic to cholinergic neurons in the medial septum, was injected into the right lateral ventricle, and electrodes were implanted bilaterally into the dorsal hippocampus of female Sprague-Dawley rats. Hippocampal electrical activity was recorded 10-14 days later from the ipsilateral (colchicine-treated) and contralateral (control) hemispheres during locomotor activity or immobility. RSA ranging from 6.3 to 8.7 Hz was evoked in both hippocampi during mobility. Following i.p. administration of an anesthetic dose of urethane, hippocampal RSA at a frequency of 4 Hz could be elicited in the control hemisphere (n = 12) of all animals by pinching the tail. RSA was absent in 6 of 9 animals in the colchicine-treated hemisphere. RSA from control and treated hemispheres persisting after urethane administration was abolished by 5 mg/kg of scopolamine, thus verifying its cholinergic nature. A decrease in the number of choline acetyltransferase (ChAT)-immunoreactive neurons in the medial septum and a depletion of acetylcholinesterase (AChE)-staining in the hippocampus were evident in the hemisphere ipsilateral to colchicine administration. These data support the septal pacemaker hypothesis of hippocampal theta-rhythm and further demonstrate the neurotoxic effect of colchicine on septo-hippocampal cholinergic neurons by the induction of a functional alteration. The selective disruption of cholinergic neurons in the medial septum by colchicine provides a means to dissociate the contribution of septal cholinergic and non-cholinergic components to hippocampal electrical activity.     

Some general anesthetics reduce serotonergic neocortical activation and enhance the action of serotonergic antagonists

In urethane-anesthetized rats, neocortical LVFA induced by 100 Hz electrical stimulation of the median raphe area or by tail pinching was completely eliminated by a combination of scopolamine (5 mg/kg, IP) and p-chlorophenylalanine (500 mg/kg/day × 3, IP), providing evidence that LVFA is dependent on cholinergic-muscarinic and serotonergic inputs to the neocortex in urethane-anesthetized as well as in freely moving rats. The serotonergic receptor antagonists ketanserin and methiothepin (1–10 mg/kg, IP) also produced a dose-dependent blockade of LVFA in urethane-anesthetized rats, and eliminated virtually all LVFA when combined with scopolamine. A combination of diethyl ether anesthesia and scopolamine completely eliminated all neocortical LVFA without additional antiserotonergic treatment, and a combination of chloral hydrate anesthesia and scopolamine similarly blocked all LVFA in about 50% of the rats tested. In the remaining chloral hydrate-anesthetized rats, the residual LVFA could be eliminated by the serotonergic antagonist ritanserin (10 mg/kg, IP). As shown previously, in nonanesthetized rats treated with scopolamine, LVFA can be maintained by a serotonergic input to the neocortex. The present data suggest that some general anesthetics reduce or completely abolish this serotonergic LVFA. Further, the serotonergic antagonists used hero exert much stronger antiserotonergic effects in rats anesthetized with urethane or chloral hydrate than in freely moving rats. Therefore, studies of serotonergic transmission or antagonist action, especially in the neocortex, in anesthetized rats may not be applicable to the waking state.     

The paradoxical effect of NMDA receptor stimulation on electrical activity of the sensorimotor cortex in freely behaving rats: analysis by combined EEG-intracerebral microdialysis.

This study was designed to determine the effects of N-methyl-D-aspartate (NMDA) receptor stimulation on the electrical activity of neocortex in freely behaving rats. Electroencephalogram (EEG) recording and intracerebral microdialysis were conducted simultaneously in the same site of the sensorimotor cortex, where the basal extracellular concentrations of aspartate and glutamate were 2.1 +/- 0.7 microM and 11.5 +/- 2.4 microM, respectively. Microdialysis with NMDA solutions (ranging from 10.0 microM to 10.0 mM) reduced the amplitude of the EEG activity and decreased the power of all frequency bands, with a virtual elimination of the high frequency waves, in a dose-dependent manner. These EEG changes were reversed after washing out the drug from the microdialysis fluid, and could be effectively antagonized with the competitive NMDA receptor antagonist DL-2-amino-5-phosphonovalerate. Remarkably, the NMDA actions were not associated with epileptiform behavioral or electrographic events. Control studies demonstrated that in the same experimental conditions, cholinergic receptor agonist carbachol caused seizures, and microdialysis with NMDA in the hippocampus readily induced epileptiform spikes. Our study shows that NMDA receptor stimulation in the rat sensorimotor cortex, although excitatory at synaptic level, can depress the local EEG activity. This may indicate that the NMDA receptor-mediated signals are processed by the neocortical network in a different way than by many other brain circuitries including hippocampus.     

Separation of hippocampal theta dipoles by partial coherence analysis in the rat

In order to separate the effect of different theta generators in the hippocampus and to characterize the pattern of relationships between them, in this study, we calculated the coherence that remains between EEG signals, recorded (1) in the stratum oriens of the CA1 region and (2) close to the hippocampal fissure in the dentate gyrus of the right or left hippocampus, after the variations, common also for a third recording site is eliminated (partialization). We found that in both anesthetized and freely moving rats, there is a selective high correlation (coherence) between theta rhythmic activities of contralateral homonymous sites of the hippocampus. The coherence between field potentials recorded in ipsilateral superficial and deep layers was eliminated when allowance was made for any of the contralateral hippocampal recordings. On the other hand, coherence between contralateral homonymous theta dipoles did not decrease when partialized by a heteronymous hippocampal EEG signal. The present results support earlier findings on multiple hippocampal theta dipoles and indicate that they can be separated using partial coherence analysis. The left and right superficial and deep dipoles oscillate as if they formed two separate systems one extending over the superficial CA1 layers on both sides and the other consisting of the left and right deep hippocampal theta dipoles. The results also suggest an important role of the commissural projections in interhemispheric theta synchronization.     

Long-term effects of basal forebrain lesions on cholinergic, noradrenergic and serotonergic markers in mouse neocortex

Lesions of basal forebrain cholinergic neurons projecting to cerebral cortex and hippocampus have recently been exploited as animal models for some of the neurochemical and behavioral deficits of Alzheimer's disease. We have observed that electrolytic lesions of cholinergic basal forebrain nuclei can lead to morphological plasticity in adult mouse cortex. In the present study, the acute and chronic sequelae of basal forebrain electrolytic lesion on cortical synaptic chemistry have been examined. In addition to choline acetyltransferase (ChAT) activity, levels of norepinephrine and of serotonin were reduced within a week after the lesion. Recovery of ChAT activity and of serotonin levels began within a month after the lesion. Serotonin type 2 receptor binding exhibited an acute reduction after the lesion in ipsilateral cortex, followed later by a chronic bilateral decrease. No significant changes in beta-adrenergic receptors were apparent at any time after the lesion despite a permanent and bilateral reduction of norepinephrine levels after the lesion. The potential significance of these results for cortical plasticity regulation and Alzheimer's disease is discussed.     

The serotonergic and noradrenergic effects of antidepressant drugs are anticonvulsant, not proconvulsant

Contrary to existing evidence, convulsant liability of the antidepressants has been attributed to noradrenergic and serotonergic increments. This is a classic case of confusing treatment effects with the manifestations of illness. In fact, the remarkable anticonvulsant effectiveness of antidepressant-induced noradrenergic and serotonergic activation has been ignored. Some antidepressant drugs such as the specific serotonin reuptake inhibitor (SSRI) fluoxetine may be devoid of convulsant liability entirely, while having distinct anticonvulsant properties. Some authorities advance the notion that the seizure predisposition of patients with epilepsy increases risks for antidepressant-induced seizures. However, evidence does not support this contention. Instead, data increasingly support the concept that noradrenergic and serotonergic deficiencies contribute to seizure predisposition. Indeed, the antidepressants have the potential to overcome seizure predisposition in epilepsy. Whereas therapeutic doses of antidepressants elevate noradrenergic and serotonergic transmission, larger doses can activate other biological processes that may be convulsant.     

Effects of serial lateral hypothalamic destruction on feeding behavior, body weight, and neocortical and hippocampal EEG activity

A comparison was made of effects of simultaneous vs. serial lateral hypothalamic lesions on (i) feeding behavior, (ii) body weight, (iii) hippocampal rhythmical slow activity, (iv) neocortical low voltage fast activity, and (v) atropine-resistant patterns of neocortical and hippocampal EEG in rats. Serial lesions separated by 30 days produced no significant sparing of function on the first three variables but produced significant sparing on the last two. The results suggest the effects may be related to anatomic and/or pharmacologic differences in the way neural systems subserving the different functions are organized.     

Evidence that serotonin mediates non-cholinergic neocortical low voltage fast activity, non-cholinergic hippocampal rhythmical slow activity and contributes to intelligent behavior

Previous research has shown that low voltage fast activity (LVFA) in the neocortex and rhythmical slow activity (RSA) in the hippocampus can result from activity in either of two ascending pathways. Activity in neurons in the basal forebrain may produce atropine-sensitive (presumably cholinergic) LVFA and RSA during both Type 1 behavior (e.g., head movement, walking) and Type 2 behavior (e.g., waking immobility, face-washing, tremor). Activity in an aminergic pathway may produce atropine-resistant LVFA and RSA during Type 1 behavior only. The role of 5-hydroxytryptamine (5-HT) in this pathway was studied in rats treated with p-chlorophenylalanine (PCPA; 500 mg/kg/day X 3, i.p.). Amine levels were measured by high pressure liquid chromatography with electrochemical detection. Brain slow wave and multi-unit activity was assessed by inspection and by a procedure of filtering and integration. PCPA treatment alone had little effect on LVFA or RSA, but following PCPA and atropine (50 mg/kg) together, both LVFA and RSA were attenuated or eliminated. Thus, atropine-resistant LVFA and RSA may be dependent on 5-HT transmission. A combination of PCPA and atropine produced a very severe deficit in performance in a simple water maze. Rats treated with this drug combination may provide an animal model of human global dementia.     

Bilateral hippocampal stimulation for intractable temporal lobe epilepsy: Impact on seizures and memory

The effect of continuous electrical stimulation of the hippocampus bilaterally on seizures and memory was assessed in two subjects with seizures from both mesial temporal lobes who were not candidates for resective epilepsy surgery. A double blind, randomized, controlled, cross‐over trial design was utilized. Two electrodes with four contacts each were implanted along the axis of the hippocampus bilaterally. Simultaneous stimulation of all electrodes contacts was either on or off during each 3‐month interval. Seizure frequency decreased by 33% in the two patients during stimulation and remained lower by 25% for the 3 months after stimulation was turned off before returning to baseline (p < 0.01). No consistent change in objective or subjective measures of memory occurred. No other adverse effects occurred. Seizure frequency is reduced both during and for a period after bilateral hippocampal stimulation, but the overall impact in this study is not as robust as has been previously reported.     

Electrical stimulation of the brain stem in freely moving rats: II. Effects on hippocampal and neocortical electrical activity, and relations to behavior

Hippocampal and neocortical slow-wave activity and behavior were recorded in freely moving rats during electrical stimulation (100 pulses/s) of sites throughout the brain stem. Stimulation at many sites (including nucleus cuneiformis; subnucleus compactus; nucleus reticularis oralis, caudalis, and medial gigantocellularis; the pontine central gray; adjacent to the midbrain central gray; centralis superior of the raphe; locus ceruleus) produced hippocampal rhythmical slow activity (RSA; theta) and neocortical low-voltage fast activity (LVFA) during behavioral immobility, especially after treatment with chlorpromazine. At slightly higher currents stimulation at most sites elicited behaviors such as walking, circling, or running, and concomitant RSA and LVFA. Atropine sulfate abolished the RSA and LVFA elicited during immobility by stimulation at most sites. The RSA and LVFA which accompanied centrally elicited locomotion was not abolished by atropine. However, exceptions to the rule that RSA and LVFA occurring during immobility are selectively abolished by atropine were provided by sites in the region of the subnucleus compactus and the nucleus of the posterior commissure. Stimulation in or near the subnucleus compactus produced primarily atropine-resistant RSA whereas stimulation in or near the nucleus of the posterior commissure produced primarily atropine-resistant LVFA. These effects could be obtained while the rats stood totally immobile. In general, the results support the idea that a diffuse ascending system from the brain stem is capable of producing two pharmacologically distinct forms of RSA and LVFA. This neurochemical specificity may be established above the brain stem. The RSA and LVFA which appear during immobility are probably dependent on cholinergic synapses. The neurotransmitter(s) required for the production of atropineresistant (movement-related) RSA and/or LVFA is unknown. It is suggested that the monoamines (noradrenaline, dopamine, and serotonin) are not critically important for the appearance of RSA or LVFA.     

Hippocampal activity, olfaction, and sniffing: an olfactory input to the dentate gyrus

Experiments in freely moving rats showed that olfactory stimuli elicit a burst of rhythmical 15-30 Hz waves in or near the hilus of the dentate gyrus but not in adjacent regions of CA1. This fast wave burst is not elicited by visual, auditory, or somatosensory inputs and is not related to motor activity. Electrical stimulation of the olfactory bulb evokes a complex potential in the hilus of the dentate gyrus but not in adjacent regions of CA1. Experiments making use of wave-triggered averaging demonstrated that there is a degree of phase-locking between (a) hippocampal RSA and sniffing or other respiratory patterns, (b) hippocampal RSA and the initiation of jumping, and (c) respiration and the initiation of jumping. An early hypothesis that the hippocampus and dentate gyrus are part of an olfacto-motor mechanism may merit re-examination.     

The effects of pumiliotoxin-B on sodium currents in guinea pig hippocampal neurons

The actions of pumiliotoxin-B, extracted from the skin of the frog Dendrobates pumilio, were examined on hippocampal slices and on acutely dissociated hippocampal neurons from the adult guinea pig. Application of 0.5-1 microM pumiliotoxin-B to hippocampal slices caused spontaneous, repetitive field discharges in the CA3 subfield. In whole-cell patch-clamp recordings of isolated CA1 and CA3 neurons, 1-2 microM pumiliotoxin-B shifted the midpoint of Na+ current activation by -11.4 +/- 1.1 mV. This shift was not dependent upon prior activation of the sodium channel. Pumiliotoxin-B did not block macroscopic Na+ inactivation but did reduce the apparent voltage-dependence of inactivation such that currents decayed faster at membrane potentials more negative than -30 mV. Single-channel recordings of sodium currents from excised membrane patches indicated that pumiliotoxin-B had little or no effect on channel closings due to entry into inactivated state(s) but did increase the rate of channel closings due to reversal of channel opening. The increase in the channel closing rate was consistent with a +8.7 mV shift in voltage sensitivity. Negative shifts in activation and positive shifts in closing rates implied a negative shift in the voltage-dependence of channel opening, suggesting that pumiliotoxin-B increases the rate of Na+ channel opening and closing in cells at rest, which could result in spontaneous activity in the neurons.     

Mapping the differential effects of procaine on frequency and amplitude of reticularly elicited hippocampal rhythmical slow activity

Hippocampal rhythmical slow activity (RSA, theta) was elicited in urethanized rats by high-frequency stimulation in the reticular formtion. The effects of procaine infusion (0.5 μL, 20% wt/vol) at various loci in the ascending system from pontine reticular formation to the medial septum/diagonal band area were investigated. It was found that procaine injected at points in the ascending system anterior to the supramamillary nucleus, in the region of the medial forebrain bundle or in the medial septum, reduced the amplitude of reticularly elicited RSA but had no effect on its frequency. Procaine injected at points in the ascending system from just anterior to the reticular formation stimulation site, up to, and including the supramamillary nucleus, reduced both the frequency and amplitude of reticularly elicited RSA. These results indicate that the frequency of reticularly elicited RSA is encoded in the supramamillary area, rather than in the medial septum/diagonal band as have previously been suggested.     

Glutamate and dopamine receptors contribute to the lateral spread of epileptiform discharges in rat neocortical slices

PURPOSE: The effects of AMPA-type glutamate receptor as well as dopamine D1 and D2 receptors on the lateral propagation of epileptiform field potentials (EFP) were studied across adjacent areas of rat neocortical tissues. METHODS: Epileptiform burst discharges were induced by superfusion of Mg(2+)-free artificial cerebrospinal fluid. Simultaneous field potential recordings of EFP were obtained from four microelectrodes placed 2-3 mm apart across coronal slices in the third layer of the neocortex. The effects of AMPA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), dopamine D1 receptor agonist SKF 81297, and dopamine D2 receptor agonist quinpirole on lateral propagation of burst discharges were investigated. RESULTS: CNQX, applied focally between recording sites, blocked rapid propagation across treated areas and resulted in the emergence of spatially separate, independent pacemakers. Focal application of SKF 81297 between recording sites increased the repetition rate of EFP, but reduced the amplitude as well as the duration of epileptic discharges. However, addition of SKF 81297 to the bath medium abolished EFP. Both local and systemic applications of quinpirole irreversibly enhanced repetition rate of epileptiform burst discharges. CONCLUSIONS: The results indicate the prerequisite of AMPA synaptic transmission for synchronized lateral propagation of Mg(2+)-free ACSF-induced epileptic activity and the modulatory effects of dopamine D1 and D2 receptors on both EFP initiation and propagation in epileptic tissues.     

Chronic peripheral inflammation,hippocampal neurogenesis,and behavior

Adult hippocampal neurogenesis is involved in memory and learning, and disrupted neurogenesis is implicated in cognitive impairment and mood disorders, including anxiety and depression. Some long-term peripheral illnesses and metabolic disorders, as well as normal aging, create a state of chronic peripheral inflammation. These conditions are associated with behavioral disturbances linked to disrupted adult hippocampal neurogenesis, such as cognitive impairment, deficits in learning and memory, and depression and anxiety. Pro-inflammatory cytokines released in the periphery are involved in peripheral immune system-to-brain communication by activating resident microglia in the brain. Activated microglia reduce neurogenesis by suppressing neuronal stem cell proliferation, increasing apoptosis of neuronal progenitor cells, and decreasing survival of newly developing neurons and their integration into existing neuronal circuits. In this review, we summarize evolving evidence that the state of chronic peripheral inflammation reduces adult hippocampal neurogenesis, which, in turn, produces the behavioral disturbances observed in chronic inflammatory disorders. As there are no data available on neurogenesis in humans with chronic peripheral inflammatory disease, we focus on animal models and, in parallel, consider the evidence of cognitive disturbance and mood disorders in human patients.     

Task-dependent effects of intracranial hippocampal stimulation on human memory and hippocampal theta power

BackgroundDespite its potential to revolutionize the treatment of memory dysfunction, the efficacy of direct electrical hippocampal stimulation for memory performance has not yet been well characterized. One of the main challenges to cross-study comparison in this area of research is the diversity of the cognitive tasks used to measure memory performance.ObjectiveWe hypothesized that the tasks that differentially engage the hippocampus may be differentially influenced by hippocampal stimulation and the behavioral effects would be related to the underlying hippocampal activity.MethodsTo investigate this issue, we recorded intracranial EEG from and directly applied stimulation to the hippocampus of 10 epilepsy patients while they performed two different verbal memory tasks – a word pair associative memory task and a single item memory task.ResultsHippocampal stimulation modulated memory performance in a task-dependent manner, improving associative memory performance, while impairing item memory performance. In addition, subjects with poorer baseline cognitive function improved much more with stimulation. iEEG recordings from the hippocampus during non-stimulation encoding blocks revealed that the associative memory task elicited stronger theta oscillations than did item memory and that stronger theta power was related to memory performance.ConclusionsWe show here for the first time that stimulation-induced associative memory enhancement was linked to increased theta power during retrieval. These results suggest that hippocampal stimulation enhances associative memory but not item memory because it engages more hippocampal theta activity and that, in general, increasing hippocampal theta may provide a neural mechanism for successful memory enhancement.