Two populations of rhythmically bursting neurons in rat medial septum are revealed by atropine

1. Previous findings, such as the sensitivity of the hippocampal theta rhythm to cholinergic manipulation, support a "pacemaker" role for the cholinergic cells of the medial septal nucleus and the vertical limb of the nucleus of the diagonal band (MSN-NDB). To explore the mechanism(s) of action of systemic antimuscarinic drugs in eliminating the theta rhythm, recordings of hippocampal EEG and rhythmic MSN-NDB neurons that fired in phase with the hippocampal theta rhythm were taken during the administration of atropine in urethane-anesthetized rats. 2. Twenty-two of 33 rhythmic MSN-NDB cells continued to burst at the theta rhythm frequency after administration of a dose of atropine (25 mg/kg iv) that was sufficient to eliminate the theta rhythm (atropine-resistant cells). The remaining 11 cells lost their rhythmic firing pattern over the same time course as the loss of the theta rhythm (atropine-sensitive cells). 3. Both types of rhythmic MSN-NDB cells could be antidromically driven from the fimbria/fornix with similar latencies (range, 0.5-4.0 ms). The extracellularly recorded spike waveforms were not useful in predicting the atropine sensitivity of a given cell. Atropine-resistant cells frequently had higher firing rates than atropine-sensitive cells, but there was sufficient overlap of the two groups to make this a poor predictor of sensitivity. 4. Cooling the fimbria/fornix reversibly eliminated the hippocampal theta rhythm, but had no effect on 21/25 rhythmic MSN-NDB cells tested. This indicates that the atropine-sensitive MSN-NDB cells do not depend on the periodic output from the hippocampus for their rhythmic firing. Recordings from pairs of rhythmic MSN-NDB cells during cooling and/or atropine administration showed unchanged phase relations at the theta rhythm frequency. In rats in which the septohippocampal system was exposed by aspirating the overlying brain tissue, direct application of atropine (10 mg/ml) to the septal nuclei reversibly eliminated the hippocampal theta rhythm. 5. The rhythmic cells of the MSN-NDB are apparently composed of at least two distinct types, both of which potentially contribute to the production of the theta rhythm in the hippocampus. Elimination of hippocampal theta rhythm after local septal atropine application suggests that the loss of rhythmic activity in the group of atropine-sensitive septal cells is sufficient for the elimination of the theta rhythm. A model of the septohippocampal connections necessary for the theta rhythm is presented.     

The Serotonin Reuptake Inhibitor Fluoxetine Suppresses Theta Oscillations in the Electroencephalogram of the Rabbit Hippocampus

Our previous studies on conscious rabbits showed that stimulation of the median cervical nucleus (MCN) decreases the extent and frequency of oscillatory theta activity in the septohippocampal system, while functional blockade of the nucleus by administration of the anesthetic lidocaine produces a stage high-frequency theta rhythm. The present study addresses the nature of the serotoninergic influences of the MCN (which also contains cells of other chemical natures) on the septohippocampal system. Experiments on conscious rabbits involved recording of the hippocampal EEG in control conditions and after microinjection of fluoxetine, a serotonin reuptake blocker which increases the levels of this transmitter in the brain. In all experiments, bilateral intracerebroventricular administration of fluoxetine hydrochloride (Sigma, St. Louis, MO; 15 microg in 5 microl of physiological saline) induced decreases in the magnitude of the hippocampal theta rhythm. In 15 of 18 (83.3%) of experiments, suppression of the oscillator activity by at least 50% of control was seen. The amplitude of the theta band in the spectral density histogram decreased by an average of 56 +/- 5.8% compared with control values (decreases in different experiments were from 7% to 90% of control p < 0.001). The latent period of these changes averaged 3.5 +/- 0.11 min (range: 2.9-4.1 min). The effect lasted 64.8 +/- 3.2 min (varying from 45.3 to 90 min in different experiments). There were no significant changes in the theta rhythm frequency, as compared with controls; this averaged 5.25 +/- 0.5 Hz (range: 4.5-6.5 Hz). The decrease in the magnitude of theta oscillations in the hippocampus after administration of fluoxetine provided evidence of the inhibitory control of rhythmic theta activity by the serotoninergic system of the brain.     

Timing of administration mediates the memory effects of intraseptal carbachol infusion

Medial septal neurons innervate the entire hippocampal formation. This input provides a potent regulation of hippocampal formation physiology (e.g. theta) and memory function. Medial septal neurons are rich in cholinergic receptors and thus are potential targets for the development of cognitive enhancers. Direct intraseptal infusion of cholinomimetics alters hippocampal physiology and can produce either promnestic or amnestic effects. Several variables (e.g. age of animal, integrity of septohippocampal circuits, task difficulty) may influence treatment outcome. We have previously demonstrated that intraseptal carbachol (12.5-125 ng) infusion immediately after the sample session of a delayed-non-match-to-sample radial maze paradigm produces a dose-dependent amnesia. The present study examined whether manipulating the timing of intraseptal carbachol infusion with respect to the sample session would alter the amnestic effect. A within-subjects design was used to examine the effect of intraseptal carbachol (125 ng/0.5 microl) in a delayed-non-match to sample radial maze task. During a sample session, rats retrieved rewards from six of 12 maze arms. At the test session (3 h later), only the alternate set contained reward and entries into the sample set arms constituted errors. Intraseptal carbachol was administered: 1) 30 min prior; 2) immediately prior; 3) immediately after and 4) 90 min after the sample session. Intraseptal carbachol prior to the sample had no effect on any index of accuracy. Infusion immediately after the sample, or delayed 90 min into the retention interval, produced an acute amnesia. These findings demonstrate that the timing of treatment is a critical variable in determining the memory effects of septohippocampal manipulations and that dynamic changes in cholinergic tone are important for memory.     

Histamine innervation and activation of septohippocampal GABAergic neurones: involvement of local ACh release

Recent studies indicate that the histaminergic system, which is critical for wakefulness, also influences learning and memory by interacting with cholinergic systems in the brain. Histamine-containing neurones of the tuberomammillary nucleus densely innervate the cholinergic and GABAergic nucleus of the medial septum/diagonal band of Broca (MSDB) which projects to the hippocampus and sustains hippocampal theta rhythm and associated learning and memory functions. Here we demonstrate that histamine, acting via H₁ and/or H₂ receptor subtypes, utilizes direct and indirect mechanisms to excite septohippocampal GABA-type neurones in a reversible, reproducible and concentration-dependent manner. The indirect mechanism involves local ACh release, is potentiated by acetylcholinesterase inhibitors and blocked by atropine methylbromide and 4-DAMP mustard, an M₃ muscarinic receptor selective antagonist. This indirect effect, presumably, results from a direct histamine-induced activation of septohippocampal cholinergic neurones and a subsequent indirect activation of the septohippocampal GABAergic neurones. In double-immunolabelling studies, histamine fibres were found in the vicinity of both septohippocampal cholinergic and GABAergic cell types. These findings have significance for Alzheimer's disease and other neurodegenerative disorders involving a loss of septohippocampal cholinergic neurones as such a loss would also obtund histamine effects on septohippocampal cholinergic and GABAergic functions and further compromise hippocampal arousal and associated cognitive functions.     

Periodic behavioral changes during hippocampal theta rhythm elicited by septal stimulation in rats

The influence of an electrical stimulation of the medial septum on the electroencephologram of the dorsal hippocampus and behavior was studied in freely moving rats. A short stimulation at 4–12 Hz frequency always induced orienting behavior (searching, rearing, sniffing, exploration) as soon as hippocampal rhythmic slow activity, or theta rhythm is elicited. An increase in stimulation intensity or in pulse duration led to an increased occurrence of theta rhythm and likewise, to an increase of orienting behavior. Both the theta rhythm and orienting behavior were dependent on the stimulation frequency.During long-lasting septal stimulation at 7 Hz, which elicited a continuous hippocampal theta rhythm, periodic behavioral changes were observed, consisting of orienting behavior alternating with grooming behavior (cleaning, washing, licking, scratching). These rhythmic periods lasted for 3–5 min.The significance of the rhythmic behavioral changes are discussed in relation to the activity of the cholinergic septohippocampal system and to rapid-eye-movement sleep and memory consolidation.     

Frontal-midline theta from the perspective of hippocampal "theta"

Electrical recordings from the surface of the skull have a wide range of rhythmic components. A major task of analysis of this EEG is to determine their source and functional significance. The hippocampal "theta rhythm" has been extensively studied in rats and its rhythmicity has recently been shown to be functionally significant, per se. Here, we use relevant aspects of the hippocampal literature to provide perspective on one of the most studied human EEG rhythms: frontal-midline theta. We review its electrographic features, localization, prevalence, age distribution, behavioural modulation (particularly in relation to working memory, spatial navigation, episodic memory, internalised attention and meditation), relationship to personality, drug interactions, neurochemical relationships, and coherence with rhythmic activity at other sites. We conclude that FM-theta, like hippocampal theta, appears to play a role in (or at least occur during) processing of memory and emotion. It is correlated with working memory and/or sustained attention; but this does not entail a role in function since clear behavioural correlates of hippocampal theta have been demonstrated that are not sensitive to hippocampal damage. FM-theta is increased by anxiolytic drug action and personality-related reductions in anxiety, whereas hippocampal theta is decreased by anxiolytic drugs. In animals, frontal theta and hippocampal theta can be phase-locked or independent, depending on behavioural state. So, the cognitive functions of FM-theta, and their relationship to hippocampal theta, are unclear and definitive evidence for functional involvement in cognitive or emotional processing is lacking. One possible solution to this problem is analysis of FM-theta in animals-provided homology can be determined. The issues of sporadicity and low incidence of FM-theta also need to be addressed in the future. Changes in functional connectivity, indicated by changes in coherence, are also a largely untapped resource. We suggest that the most hopeful path to assessing the functions of FM-theta will be through the use of drugs, and the variation of their effects depending on baseline levels of FM-theta. Finally, we review some theories of theta function. Despite the apparent richness of the current data, we conclude that it is difficult (and may ultimately be impossible) to formulate a theory that attributes a specific cognitive function to FM-theta. However, the theories share some general computational assumptions and these should be a useful guide to future work and, ultimately, a definite theory of the function or functions of FM-theta.     

Sensory Responses of Neurons in the Medial Septal Area in Conditions of Modulation of Theta Activity Using the Alpha-2-Adrenoreceptor Agonist Clonidine

Our previous studies on conscious rabbits showed that administration of the alpha-2-adrenoreceptor agonist clonidine induces dose-dependent changes in theta oscillations in the septohippocampal system. Low doses of clonidine suppressed theta activity, while high doses produced significant potentiation. It was suggested that the different effects of clonidine might be associated with differences in the sensitivities of pre- and postsynaptic alpha-2-adrenoreceptors to clonidine, this agent being a pure agonist of noradrenaline when used at high doses. It was suggested that functional synergism occurs between the activatory reticular formation and the noradrenergic system of the locus ceruleus in controlling the theta rhythm. The present study was performed to identify the nature of the responses of sensory neurons in the medial septal region in conditions of alterations in the magnitude of the theta rhythm induced by different doses of clonidine. Low and high doses of the agonist given bilaterally into the lateral ventricles were found to have different effects on the sensory responses of neurons in the medial septal region. Injection of small clonidine doses (0.5 microg in 5 microl into each lateral ventricle), which decrease theta activity, was found to lead to weakening of activatory processes and enhancement of inhibitory processes in the medial septal region. The number of activatory responses decreased significantly and persisting responses were significantly weakened; inhibitory responses, conversely, were seen more frequently and were significantly more marked. Administration of high clonidine doses (5 microg in 5 microl), which produce sharp increases in theta oscillations, led to significant reductions in the reactivity of cells in the medial septal region to sensory stimuli (from 76.8% in controls to 45% after clonidine), regardless of the nature of the initial responses. Persisting activatory and inhibitory responses were in most cases less marked than the initial responses. These results suggest that alpha-2-adrenoreceptors are involved in controlling the sensory reactivity of neurons in the medial septal region. The impairment of the normal processing of sensory stimuli seen during the continuous generation of rhythmic activity provoked by injection of large clonidine doses supports the role of the theta rhythm in the septohippocampal system as an active filter during the processing and recording of information.     

Brainstem sites for the carbachol elicitation of the hippocampal theta rhythm in the rat

The effects of brainstem microinjections of carbachol on the hippocampal theta rhythm were examined in urethane anesthetized rats. The two most effective theta-eliciting sites with carbachol were the nucleus pontis oralis (RPO) and the acetylcholine-containing pedunculopontine tegmental nucleus (PPT) of the dorsolateral pontine tegmentum. RPO injections generated theta at mean latencies of 38.5±70.8 s and for mean durations of 12.9±5.1 min. Five of seven RPO injections gave rise to theta virtually instantaneously, i.e., before the completion of the injection. PPT injections generated theta at mean latencies of 1.7±1.1 min and for mean durations of 11.9±6.0 min. Injections rostral or caudal to RPO in the caudal midbrain reticular formation (RF) or the caudal pontine RF (nucleus pontis caudalis) generated theta at considerably longer latencies (generally greater than 5 min) or were without effect. Medullary RF injections essentially failed to alter the hippocampal EEG. The finding that theta was produced at very short latencies at RPO suggests that RPO, the putative brainstem source for the generation of theta, is modulated by a cholinergic input. The further demonstration that theta was also very effectively elicited with PPT injections suggests this acetylcholine-containing nucleus of the dorsolateral pons may be a primary source of cholinergic input to RPO in the generation of theta. The hippocampal theta rhythm is a major event of REM sleep. The present results are consistent with earlier work showing that each of the other major events of REM sleep, as well as the REM state, are cholinergically activated at the level of the pontine tegmentum.     

Action potentials and relations to the theta rhythm of medial septal neurons in vivo

The influence of the medial septal nucleus and the nucleus of the diagonal band of Broca (MS-DB) on the hippocampal theta rhythm includes both cholinergic and γ-aminobutyric acid (GABAergic) components. To understand the intrinsic septal interactions and the separate contributions of the cholinergic and GABAergic septohippocampal neurons to the theta rhythm in behaving animals, it is essential to be able to identify these two classes from extracellular recordings. Here the durations of extracellularly recorded action potentials are compared with the other characteristics of the neurons. Extracellular recordings were taken from neurons of the MS-DB both in freely moving rats (114 cells) and in urethane-anesthetized rats (112 cells). These were compared with intracellular recordings taken from MS-DB neurons in urethane-anesthetized rats (58 cells). Hippocampal EEG was recorded from above the CA1 pyramidal cell layer (CA1 theta) and near the hippocampal fissure (dentate theta) to compare the firing phase across cells. Here it is shown that two major types of rhythmically bursting cells in the MS-DB that had been distinguished previously in intracellular recordings in vivo are also separable in extracellular recordings in vivo on the basis of the durations of their action potentials. In both awake and anesthetized rats the main properties of the two cell types were found to differ: firing rate, phase-relation to the hippocampal theta rhythm and sensitivity of their rhythmicity to blockade of muscarinic transmission. As was previously shown for intracellular recordings in anesthetized rats, it is shown here that in awake rats, too, the more rapidly firing brief-spike (putative GABAergic) cells fired with highest probability on the negative phase of the dentate theta, whereas the more slowly firing long-spike (putative cholinergic) cells fired mostly on the positive phase. Previous work showed that in intracellular recordings from anesthetized rats the rhythmic firing of most brief-spike cells was still retained even during muscarinic blockade, but that of most long-spike cells was lost. Here we also report a recategorization according to spike duration of existing extracellular recordings taken from anesthetized rats, confirming the above observation with much larger numbers of cells. Three additional major new findings are also reported here. (1) In awake rats, muscarinic blockade has relatively little effect on either cell type. (2) Under anesthesia, the firing rates of both cell types are lower than in awake rats, but the effect is greater on the long-spike cells, where the anesthesia also reduces the rhythmicity of the cell firing. (3) Rhythmicity of the putative GABAergic cells is also retained after local injection of GABA-A antagonist, whereas that of the putative cholinergic cells is eliminated. We conclude that either systemic muscarinic blockade or urethane anesthesia alone have relatively little effect on neurons in the defined above MS-DB, but a combination of the two has profound effects on the rhythmicity of the cholinergic cells, largely sparing the GABAergic cells. Taken together, the results suggest that generation of theta rhythm requires a background of excitatory influences on the hippocampus (that can be maintained by either muscarinic or glutamatergic inputs) in combination with the phasic disinhibitory action mediated by the GABAergic MS-DB projection. They also provide additional support for the notion that the phasic activity in local collaterals of GABAergic MS-DB cells contributes to the phasic modulation of the firing of cholinergic septohippocampal neurons. Received: 13 October 1998 / Accepted: 15 March 1999     

Ventral hippocampal dopamine D1 and D2 systems and spatial working memory in rats

The hippocampus has long been known to be important for memory function. However, the involvement of hippocampal dopamine systems with memory has received little attention. In the current study, dopamine D1 and D2 hippocampal receptor system involvement with memory was assessed in female Sprague-Dawley rats by local infusion of D1 and D2 agonists and antagonists into the ventral hippocampus. Working memory performance was assessed on the radial-arm maze. Neither the D1 agonist dihydrexidine (1.1-10 microg/side) nor the D1 antagonist SCH 23390 (0.19-1.67 microg/side) was effective in significantly altering radial-arm maze choice accuracy. In contrast, there were significant and opposite effects of D2 agonist and antagonist treatments. The D2 agonist quinpirole caused a significant (P<0.05) dose-related improvement in choice accuracy over a dose range of 1.1-10 microg/side. In a complementary fashion, the D2 antagonist raclopride caused a significant (P<0.05) dose-related choice accuracy deficit over a range of 0.19-1.67 microg/side. This study provides clear evidence that hippocampal D2 activity is positively related to working memory performance, while evidence for D1 systems is less compelling. Dopamine D2 receptors in the ventral hippocampus were shown to have important influences on spatial working memory. In a consistent pattern of effects ventral hippocampal infusion of the D2 agonist quinpirole improved working memory performance in the radial-arm maze, while ventral hippocampal infusion of the D2 antagonist raclopride impaired performance.     

Preservation of a functionally intact neuronal network after global ischemia

Although conventional histological techniques demonstrated preservation by drug treatment of neuronal perikarya in the hippocampal CA1 region in animals subjected to short periods of transient ischemia, uncertainty exists regarding whether the anatomical connections with these neurons are intact and functional. The hippocampal theta rhythm is dependent upon intact connections to the CA1 pyramidal neurons and is a useful predictor of functional hippocampal integrity. Hippocampal theta was quantitated by power spectral analysis in rats subjected to 30 min of 4-vessel occlusion (4-VO) and treatment with the neuroprotective antioxidant, LY231617. The 4-VO destroyed CA1 neurons and reduced the amount of theta, however, LY231617 protected CA1 neurons histologically and totally preserved the hippocampal theta rhythm. We conclude that histological preservation is indicative of functional integrity.     

Medial septal beta-amyloid 1-40 injections alter septo-hippocampal anatomy and function

Degeneration of septal neurons in Alzheimer's disease (AD) results in abnormal information processing at cortical circuits and consequent brain dysfunction. The septum modulates the activity of hippocampal and cortical circuits and is crucial to the initiation and occurrence of oscillatory activities such as the hippocampal theta rhythm. Previous studies suggest that amyloid beta peptide (Abeta) accumulation may trigger degeneration in AD. This study evaluates the effects of single injections of Abeta 1-40 into the medial septum. Immunohistochemistry revealed a decrease in septal cholinergic (57%) and glutamatergic (53%) neurons in Abeta 1-40 treated tissue. Additionally, glutamatergic terminals were significantly less in Abeta treated tissue. In contrast, septal GABAergic neurons were spared. Unitary recordings from septal neurons and hippocampal field potentials revealed an approximately 50% increase in firing rates of slow firing septal neurons during theta rhythm and large irregular amplitude (LIA) hippocampal activities and a significantly reduced hippocampal theta rhythm power (49%) in Abeta 1-40 treated tissue. Abeta also markedly reduced the proportion of slow firing septal neurons correlated to the hippocampal theta rhythm by 96%. These results confirm that Abeta alters the anatomy and physiology of the medial septum contributing to septo-hippocampal dysfunction. The Abeta induced injury of septal cholinergic and glutamatergic networks may contribute to an altered hippocampal theta rhythm which may underlie the memory loss typically observed in AD patients.     

Modulation of theta rhythmicity in the medial septal neurons and the hippocampal electroencephalogram in the awake rabbit via actions at noradrenergic alpha2-receptors

The modulation of the firing discharge of medial septal neurons and of the hippocampal electroencephalogram (EEG) mediated by actions on alpha2-adrenoreceptors (ARs) was investigated in awake rabbits. Bilateral i.c.v. infusion of a relatively low dose (0.5 microg) of the alpha2-AR agonist clonidine produced a reduction in the theta rhythmicity of both medial septal neurons and the hippocampal EEG. In contrast, a high dose of clonidine (5 microg) increased the percentage and degree of rhythmicity of theta bursting medial septal neurons as well as the theta power of the hippocampal EEG. On the other hand, administration of alpha2-AR antagonist idazoxan produced the opposite dose-dependent effect. While a low dose of the antagonist (20 microg) produced an increase in both the theta rhythmicity of medial septal neurons and the theta power of the hippocampal EEG, a high dose (100 microg) caused a reduction of theta rhythmicity in both the medial septum and hippocampus. These results suggest that low doses of alpha2-ARs agents may act at autoreceptors regulating the synaptic release of noradrenaline, while high doses of alpha2-ARs drugs may have a predominant postsynaptic action. Similar results were observed after local injection of the alpha2-AR drugs into the medial septum suggesting that the effects induced by the i.c.v. infusion were primarily mediated at the medial septal level. We suggest that noradrenergic transmission via the postsynaptic alpha2-ARs produces fast and strong activation of the septohippocampal system in situations that require urgent selective attention to functionally significant information (alert, aware), whereas the action via the presynaptic alpha2-ARs allows a quick return of the activity to the initial level.     

Changes in ensemble activity of hippocampus CA1 neurons induced by chronic morphine administration in freely behaving mice

The hippocampus plays an important role in the formation of new memories and spatial navigation. Recently, growing evidence supports the view that it is also involved in addiction to opiates and other drugs. Theoretical and experimental studies suggest that hippocampal neural-network oscillations at specific frequencies and unit firing patterns reflect information of learning and memory encoding. Here, using multichannel recordings from the hippocampal CA1 area in behaving mice, we investigated the phase correlations between the theta (4-10 Hz) and gamma (40-100 Hz) oscillations, and the timing of spikes modulated by these oscillations. Local field potentials and single unit recordings in the CA1 area of mice receiving chronic morphine treatment revealed that the power of the theta rhythm was strongly increased; at the same time, the theta frequency during different behavioral states shifted markedly, and the characteristic coupling of theta and gamma oscillations was altered. Surprisingly, though the gamma oscillation frequency changed, the power of gamma lacking theta did not. Moreover, the timing of pyramidal cell spikes relative to the theta rhythm and the timing of interneuron spikes relative to the gamma rhythm changed during chronic morphine administration. Furthermore, these responses were impaired by a selective D1/D5 receptor antagonist intra-hippocampus injection. These results indicate that chronic morphine administration induced the changes of ensemble activity in the CA1 area, and these changes were dependent on local dopamine receptor activation.     

Repeated blockade of GABAA receptors in the medial septal region induces epileptiform activity in the hippocampus

Changes in the activity of putative interneurons of the stratum oriens of the hippocampus and hippocampal EEG after the delivery of the GABAA receptor antagonist bicuculline (1.0nmol/1microl) to the medial septal region were investigated in awake rabbits. The injection of bicuculline produced a sharp increase in the firing rate in 94.3% of hippocampal cells. The effect of bicuculline on the neuronal theta rhythmicity depended on the number of injections. The first five daily infusions decreased the theta activity in 85.7% of cells. On the fourth to fifth experimental days paroxysmal discharges and 8-15Hz oscillations were recorded in the hippocampal EEG. Six to seven further daily bicuculline injections following a brief diminution of theta activity produced a sharp augmentation of theta oscillations in 78.9% of cells and provoked seizures. Immediately before seizures, stabilization of theta bursts and an increase in burst frequency was usually observed in putative interneurons. During seizures, neuronal rhythmic activity was either disordered and then turned into seizure discharges or was inhibited, partially or completely. In the hippocampal EEG, the power of theta rhythm before seizures usually strongly increased compared with controls. Injection of the GABAA agonist muscimol (30nmol/1microl) 15min before bicuculline infusion prevented the development of seizures. These findings suggest that the interplay between septal neurons via GABAA receptors is critical in the tuning of septal output signals that insure generation of natural theta rhythm as well as adequate functioning of the hippocampus.     

Spontaneous activity and sensory responses of hippocampal neurons during persistent theta-rhythm evoked by median raphe nucleus blockade in rabbit

Spontaneous activity and responses to sensory stimuli were analysed in the hippocampal CA1 neurons of chronic unanesthetized rabbits before and after reversible functional blockade of the median raphe nucleus and medial septal area by local microinjections of anesthetic lidocaine. This evoked, correspondingly, persistent theta rhythm and its complete blockade for about 30 min. The results were compared to the neuronal data obtained earlier in the experiments with cholinergic drugs modulating expression of theta rhythm. Intra-median raphe nucleus injection of lidocaine evoked uniform increase of discharge rate in the hippocampal neurons with low and high spontaneous activity. Theta modulation of neuronal activity had increased regularity and frequency (by 0.5-2.0 Hz) and appeared in additional group of the neurons simultaneously with expression of persistent theta in the hippocampal electroencephalogram. Sensory responsiveness of the hippocampal neurons was drastically decreased (45% of the responses preserved). Reactions of all types were blocked, diminished, or inverted, but inhibitory responses were the most severely affected. Injection of lidocaine into medial septal area also blocked all brain stem afferents ascending to the hippocampus via medial septal area, thus, totally depriving hippocampus of brainstem-septal input. However, besides the total absence of theta modulation, spontaneous activity in majority of neurons was not significantly changed. Responsiveness to sensory stimuli also remained relatively high (77% of the responses preserved); on-effects were especially resistant to medial septal area blockade. Comparison of spontaneous and evoked activity in two theta states (physostigmine and median raphe nucleus blockade) revealed striking similarity of all characteristics, which suggested that theta-suppressing influences of median raphe nucleus (presumably serotonergic) are realized primarily through the control of cholinergic septo-hippocampal theta-generating mechanism. However, as the frequency of theta rhythm does not depend on it, an additional effect of disinhibition of activating reticular formation by the median raphe nucleus suppression is suggested. The data confirm that theta rhythm may be regarded as active filter in the information processing by the hippocampal neurons.     

Modulation of the reaction of hippocampal neurons to sensory stimuli by cholinergic substances

The influences of increasing endogenous acetylcholine (eserine) and its blockade (scopolamine) on the effects of sensory stimuli were analyzed through the extracellular recording of the activity of individual hippocampal neurons of awake rabbits. An increase in the level of acetylcholine, accompanied by the appearance of stable theta rhythm, leads to a substantial decrease in the reactivity of neurons, the suppression, attenuation, and inversion of the majority of inhibitory reactions and of a substantial proportion of activational reactions including on-responses of a specific type. At the same time, a limited group of activational reactions is intensified and extended against the background of eserine. Scopolamine, which blocks theta rhythm, does not change or intensifies inhibitory and some activational reactions, including on-responses. Tonic reactions are shortened; however, their gradual extinction disappears. The effects described are preserved in the hippocampus in the presence of basal undercutting of the septum which eliminates ascending brainstem pathways. These data make it possible to draw the conclusion that, under normal conditions, a new (significant) sensory stimulus elicits in the hippocampus an initial stoppage (reset) of activity with the coordinated triggering of theta rhythm and the passage against this background of signals along the cortical input in a specific phase relationship to it. The period of theta modulation switched on by the signal fosters its recording and the limitation of the passage of subsequent, interfering signals. The septohippocampal influences may thus support the mechanism of selective attention, as a necessary precondition for memory. This study carried out with the financial support of the Russian Basic Research Fund (Project No. 93-04-21907). Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino. Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 45, No. 1, pp. 118–131, January–February, 1995.     

GABAergic control of the ascending input from the median raphe nucleus to the limbic system

The median raphe nucleus (MRN) is the primary source of serotonergic afferents to the limbic system that are generally considered to suppress hippocampal theta oscillations. GABA receptors are expressed in the MRN by serotonergic and nonserotonergic cells, including GABAergic and glutamatergic neurons. This study investigated the mechanisms by which the fluctuating GABA tone in the MRN leads to induction or suppression of hippocampal theta rhythm. We found that MRN application of the GABA(A) agonist muscimol (0.05-1.0 mM) or GABA(B) agonist baclofen (0.2 mM) by reverse microdialysis had strong theta promoting effects. The GABA(A) antagonist bicuculline infused in low concentrations (0.1, 0.2 mM) eliminated theta rhythm. A short period of theta activity of higher than normal frequency preceded hippocampal desynchronization in 46% of rats. Bicuculline in larger concentrations (0.5, 1.0, 2.0 mM) resulted in a biphasic response of an initial short (<10 min) hippocampal desynchronization followed by stable theta rhythm that lasted as long as the infusion continued. The frequency and amplitude of theta waves were higher than in control recordings and the oscillations showed a conspicuous intermittent character. Hippocampal theta rhythm elicited by MRN administration of bicuculline could be completely (0.5 mM bicuculline) or partially (1.0 mM bicuculline) blocked by simultaneous infusion of the GABA(B) antagonist CGP35348. Our findings suggest that the GABAergic network may have two opposing functions in the MRN: relieving the theta-generators from serotonergic inhibition and regulating the activity of a theta-promoting circuitry by the fluctuating GABA tone. The two mechanisms may be involved in different functions.     

Increase of hippocampal acetylcholine release at the onset of dark phase is suppressed in a mutant mice model of evening-type individuals

We have previously reported that clock mutant mice on Jcl:ICR background show about 2-h delayed circadian profiles in body temperature, spontaneous activity and sleep-wake rhythm, and that they appear to be useful as a model of evening-type of individual. Hippocampal acetylcholine (ACh) release which is positively correlated with attention, learning and memory shows a circadian variation. In this study, changes in hippocampal ACh release in transitional phase from light (rest) to dark (active) period in clock mutant mice were monitored using an in vivo microdialysis method. Compared with wild mice, the increase in hippocampal ACh in the first 2 h of the active period in the mutant mice was suppressed in parallel with peak frequency in electroencephalogram theta rhythm. The molecular basis of the circadian system appears to have a strong effect on hippocampal cholinergic function, and is probably associating with individual temporal differences in voluntary behavior, cognition, learning and/or memory performance.     

Long-term working memory in the rat: effects of hippocampally applied anisomycin

The extent to which protein synthesis is involved in working memory was investigated with the protein synthesis inhibitor anisomycin (ANI). Rats were trained to perform accurately on a 12-arm radial maze when delays of 240 min were interposed between choice 6 and choice 7. Bilateral hippocampal cannulas were then implanted. Accuracy on choices 7-12 was studied when ANI or saline was injected either 30 min before choice 1 or 5-10 min after choice 6 (Experiment 1). Pretrial injection of ANI significantly impaired performance following the 240-min delay, whereas ANI injected during the delay had no such effect. In Experiments 2 and 3, the ANI-induced amnesia was replicated, and the temporal course of development of the amnesia was determined. Pretrial administration of ANI did not significantly affect retention after a 2-min delay but did produce amnesia after delays of 15 min or longer. These data suggest that protein synthesis is important for the formation of temporary memories, provided the retention interval is long enough. It is suggested that working memory includes both short-term and long-term components. Protein synthesis appears to be important for formation of the long-term component, but not the short-term component, of working memory.