Electrophysiological analysis of the hippocampal-septal projections: II. Functional characteristics

Summary 1. The septum and dorsal hippocampus were exposed in rats by the ablation of the overlying cortex and corpus callosum. The hippocampal efferent system, fornix and fimbria, were stimulated with either single or paired shocks and field potentials, extra- and intra-cellular unitary potentials were recorded in the lateral septal nucleus. 2. The double-shock pairing within the same hippocampal system resulted in facilitation of the monosynaptic test field response. 3. Pairing across systems resulted in long-term suppressions of the mono-synaptic test field response. 4. IPSPs and lack of convergence of excitatory input are responsible for the long-term suppression. 5. It is suggested that the IPSPs are mediated via interneurons, and that some of which at least are activated recurrently. 6. Hippocampal stimulation suppresses the amygdaloid evoked field response for very long periods, often beyond 1000 msec.This study was supported by U.S. Public Health Service Grants RR 5384 and NB 00405.     

Direct projections from Ammon's horn to the septum in the cat

Summary Direct projections from Ammon's horn to the septum were studied in the cat by the anterograde tracing method after injecting WGA-HRP (wheat germ agglutinin-horseradish peroxidase conjugate) into Ammon's horn. The results were further confirmed by the retrograde WGA-HRP method after injecting WGA-HRP into the septum. Pyramidal neurons in fields CA1, CA2 and CA3 were observed to send their axons ipsilaterally to the lateral septal nucleus; the septal parts of the hippocampus sent projection fibers to the dorsomedial portions of the lateral septal nucleus via the medial aspects of the subcallosal fornix, while the hippocampal regions successively more proximal to the temporal pole sent projection fibers to progressively more ventrolateral portions of the lateral septal nucleus via more lateral aspects of the subcallosal fornix. It was also found that the septal parts of fields CA1, CA2 and CA3 sent projection fibers bilaterally to the dorsomedial aspects of the lateral septal nucleus. Field CA4 appeared to send projection fibers only sparsely, if at all, to the medial septal nucleus. The rudimentary parts of the hippocampal formation, taenia tecta and indusium griseum, were found to have reciprocal ipsilateral connections with the dorsal portions of the lateral septal nucleus.     

Interaction of age and sex in sympathetic axon ingrowth into the hippocampus following septal afferent damage

Summary Damage to the medial septal nucleus or to the fimbria/fornix in the adult rat elicits sprouting of vascular sympathetic fibers into the deafferented regions of the hippocampal formation. The following study examines the effects of developmental stage and sex on this sprouting phenomenon using both fluorescence histochemistry and high affinity uptake of noradrenaline. We find that (1) the sprouting, which is reduced in adult and juvenile males relative to females, is equivalent in the two sexes after transections at postnatal day 3, and (2) the period of maximum ingrowth is sexually regulated, occurring near postnatal day 3 in the male and postnatal day 13 in the female.     

Organization of identified fiber tracts in the rat fimbria-fornix: an anterograde tracing and electron microscopic study

The fimbria is a major route for afferent and efferent fibers of the hippocampal formation. However, little is known about the intrinsic organization of the fimbria-fornix complex. In this study, the anterograde tracer Phaseolus vulgaris-leucoagglutinin (PHAL) was used to analyze the ultrastructure and topography of identified fiber tracts within the fimbria-fornix. Septo-hippocampal fibers are loosely distributed throughout the fimbria-fornix. Commissural fibers cross the midline in the ventral hippocampal commissure and form a tight fiber bundle in the fimbria. Crossed entorhino-hippocampal fibers cross the midline in the ventral hippocampal commissure rostral to the commissural fiber bundle, and crossed entorhino-entorhinal fibers pass through the dorsal hippocampal commissure. This suggests a topographical organization of fiber tracts within the fimbria-fornix that reflects the laminar organization of the hippocampal target structure: fibers of the diffusely terminating septohippocampal projection are loosely distributed throughout the fimbria-fornix, while those projections that are known to terminate in specific laminae of the hippocampal formation (commissural projection, crossed entorhino-hippocampal projection) form fiber bundles within the fimbria and the ventral hippocampal commissure.Abbreviations A Astrocyte - CA1, CA3 hippocampal subfields - CC corpus callosum - D dendrite - DG dentate gyrus - DHC dorsal hippocampal commissure - Fi fimbria - LS lateral septal area - LV lateral ventricle - O oligodendrocyte - SFO subfornical organ - VHC ventral hippocampal commissure     

The role of the entorhinal cortex in two forms of spatial learning and memory

It is generally acknowledged that the rodent hippocampus plays an important role in spatial learning and memory. The importance of the entorhinal cortex (ERC), an area that is closely interconnected anatomically with the hippocampus, in these forms of learning is less clear cut. Recent studies using selective, fibre-sparing cytotoxic lesions have generated conflicting results, with some studies showing that spatial learning can proceed normally without the ERC, suggesting that this area is not required for normal hippocampal function. The present study compared cytotoxic and aspiration ERC lesions with both fimbria fornix (FFX) lesions and sham-operated controls on two spatial learning tasks which have repeatedly been shown to depend on the hippocampus. Both groups of ERC lesions were impaired during non-matching-to-place testing (rewarded alternation) on the elevated T-maze. However, neither of these lesions subsequently had any effect on the acquisition of a standard spatial reference memory task in the water maze. FFX lesions produced a robust and reliable impairment on both of these tasks. A second experiment confirmed that cytotoxic ERC lesions spared water maze learning but disrupted rewarded alternation on the T-maze, when the order of behavioural testing was reversed. These results confirm previous reports that ERC-lesioned animals are capable of spatial navigation in the water maze, suggesting that the ERC is not a prerequisite for normal hippocampal function in this task. The present demonstration that ERC lesions disrupt non-matching-to-place performance may, however, be consistent with the possibility that ERC lesions affect attentional mechanisms, for example, by increasing the sensitivity to recent reward history.     

Quantitative analysis of the topographical organization of olivocerebellar projections in the rat

The inferior olivary projection to anterior and posterior regions of the cerebellar vermis was studied in the rat using the retrograde fluorescent double-labelling technique in combination with a computerized data collection system. A technique for quantifying and statistically analysing the point-to-point (or nucleotopic) organization of the projection is presented. It was found that one group of neurones in the medial accessory olivary nucleus projected to cerebellar lobules 6 or 7, and second group projected to lobules 4 or 5, while a third sent collaterals to both areas. In the rostral part of the nucleus these three groups were spatially separated (P less than 0.05), whereas caudally they were intermingled. It is concluded that a double-labelling method has distinct advantages over a single-tracer technique (such as horseradish peroxidase) in the investigation of the topographic organization of neuronal projections. Quantitative analysis is used here to overcome the problems presented by collateralization, inter-animal variation and the accurate representation of results from a series of animals.     

Septo-hippocampal connections and the hippocampal theta rhythm

Summary Recordings were made of spontaneous hippocampal theta activity in free-moving rats, before and after a variety of lesions. Three recording sites were used to monitor activity in the dorsal hippocampus, the ventral hippocampus, or close to the site of the hippocampal flexure. Electrolytic lesions were made in the medial septal area or the dorso-lateral septal area; surgical transections were made of the fimbria or dorso-medial area of the fornix. Following lesions restricted to the medial septal area, theta was abolished throughout the hippocampus; after lesions restricted to the dorso-lateral septal area theta was retained. Fimbria lesions abolished theta in the ventral, but not the dorsal hippocampus; dorso-medial fornix lesions abolished it in the dorsal, but not the ventral, hippocampus. In some subjects the hippocampal formation was subsequently stained for cholinesterase: cholinesterase staining loss was generally associated with theta loss, but this was not clear at the flexure recording site. It was confirmed that theta is dependent upon the integrity of the medial septal area. It was concluded that damage to hippocampal afferents from the septum does abolish theta, while damaging the feedback efferents does not.     

Electrophysiological analysis of the hippocampal projections to the entorhinal area

Responses evoked in the entorhinal area by impulse volleys originating in the ipsilateral hippocampus were analysed in the guinea-pig by means of field potential analysis. Perforant path volleys, synaptically elicited by stimulation of the dorsal psalterium of one side, were used to activate the hippocampal lamellar circuit of the same side and, through interhippocampal impulses, the hippocampal pyramidal neurons of the contralateral side. Discharge of the hippocampal pyramidal neurons was followed by a response, a fast negative deflection preceded and followed by slow waves, in the dorsal third of the ipsilateral entorhinal area. Laminar distribution of the fast negative deflection and of the time-locked unit activity suggested that excitatory synaptic effects followed by neuron discharge were generated in neurons of layers VI-II of the entorhinal area. The increasing latency of the fast negative deflection and of unit firing over the cortical depth suggested that these synaptic effects were generated in temporal sequence, going from layer VI to layer II. The entorhinal response disappeared after a lesion at the caudal border of the hippocampus interrupting the caudally-directed hippocampal efferents. The anatomy of the hippocampal and subicular projections to the entorhinal area in the guinea-pig, together with electrophysiological data obtained in recordings from the ipsilateral subiculum, suggested that the hippocampal impulses were relayed to layers VI-V of the entorhinal area by the subiculum. The delayed activation of layers IV-II was possibly mediated by intracortical connections. Double-shock experiments showed that impulses of hippocampal origin inhibited the response to dorsal psalterium volleys of entorhinal neurons giving origin to perforant path fibers. The data show that the hippocampal output activates the deep layers of the entorhinal area from which it is possibly relayed to numerous cortical and subcortical regions. Moreover, the inhibitory effects exerted on neurons originating perforant path fibers give evidence of a negative feedback control system operating in the hippocampal region.     

培养大鼠海马胆囊收缩素阳性神经元的电生理学特性

目的 探索海马内胆囊收缩素(CCK) 阳性（CCK⁺ ）神经元的电生理学特性。方法 用整合有CCK启动子-绿色荧光蛋白(GFP)的腺病毒对培养海马CCK⁺神经元进行标记,应用全细胞膜片钳技术对标记CCK⁺神经元的电生理学特性进行研究。结果 CCK⁺神经元的胞体长径为(22.85±0.77)μm,短径为(16.21±0.42)μm (n=20),静息电位 (RMP)为(-55.90±1.30)mV,膜电容 (Cm)为(45.77±2.06)pF,膜阻抗 (Rm)为(711.00±46.69)MΩ; CCK+神经元500 ms内100 pA去极化电流诱发动作电位的发生频率为(26.17±3.41)Hz (n=12); CCK⁺神经元的自发抑制性突触后电流的发生频率［(5.26±0.71)Hz, n=7］显著高于自发兴奋性突触后电流的发生频率［(3.24±0.62)Hz, n=6］。 结论 CCK⁺神经元的相关电生理学特征可能与其在中枢神经系统内重要的调节功能相关。     

The organization of the fimbria, dorsal fornix and ventral hippocampal commissure in the rat

Summary The morphological organization of the two principal efferent pathways of the hippocampal formation — the fimbria and the dorsal fornix — and of the interhippocampal fibers that cross in the ventral hippocampal commissure, has been studied autoradiographically following injections of ³H-amino acids into the relevant cytoarchitectonic fields. To a considerable extent the rostrally-directed fibers from each cytoarchitectonic field remain topographically segregated within the fimbria or dorsal fornix, their locations being determined by the relative levels at which they join these bundles. Thus, the fibers from the subicular region tend to be aggregated along the ependymal border of the fimbria while those from the hilar region of the dentate gyrus are medially-placed along its pial margin. Similarly, the efferents from the septal and temporal parts of the various fields are distributed along the proximo-distal dimension of the fimbria, from its base (adjacent to field CA₃) to its tip. The dorsal fornix contains fibers from the septal (dorsal) part of the subiculum and field CA₁; although there appears to be some overlap, the subicular efferents tend to be more medially placed while those from CA₁ are further lateral. The topographic organization of the fibers in the fimbria is reflected in the arrangement of the crossed components in the hippocampal commissure. Those arising at more temporal levels cross most rostrally while those from the septal pole of field CA₃ cross in the caudal part of the commissure; those from the hilar region of the dentate gyrus cross most ventrally, while those from the more lateral parts of field CA₃ (CA_3b and CA_3a) cross at successively more dorsal levels.We estimate that the fimbria contains about 900,000 fibers of which approximately 700,000 are myelinated (with internal diameters ranging between 0.1 and 2.8m; mean 0.78 m) and just over 200,000 are unmyelinated (with diameters between 0.1 and 0.8 m; mean 0.24 m).Supported by Grants EY-01255, NS-10943 and NS-13267 from the National Institutes of Health, and by a grant from the Sloan Foundation. We should like to thank Mrs. LueVern Bell and Mrs. Pamela Seyer for their excellent histological work and Mrs. Mary Murphy for secretarial help     

Medial septal projections to the dentate gyrus of the rat: electrophysiological analysis of distribution and plasticity

Summary Previous electrophysiological experiments in rabbits have suggested that medial septal stimulation activates dentate granule cells and evokes an associated negative field response at the granule cell layer, without an associated dendritic response. Anatomical studies have suggested that septal input to the granule cells may be to stratum moleculare, or close to the cell layer, or may not exist at all. The present experiments confirmed in rats anaesthetised with urethane that medial septal stimulation elicits single action potentials from cells in the granule layer. The associated negative field potential was maximal in the granule cell layer and there was no sign of a separate dendritic potential. The fibres responsible for this potential travel to the dorsal hippocampus in the fornix superior rather than the fimbria, taking the same course as the fibres which contribute to the dense cholinesterase staining just above the granule cell layer. Stimulation at 100 Hz for 1 s of either medial septal, or perforant path, input to the dentate granule cell layer produced long term potentiation of the subsequent evoked field responses to the stimulated pathway. The responses to the non-stimulated pathway were unchanged. Paired pulse stimulation produced both homosynaptic and heterosynaptic potentiation. These data suggest that medial septal input synapses close to granule cell bodies and produces a negative field potential which is a combination of dendritic and population spike potentials. Medial septal input also appeared to produce direct activation of hilar neurones, some of which may be basket cells or other interneurones. The data also show that long term potentiation is specific to this input, perhaps dependent on presynaptic mechanisms. Paired pulse potentiation, at least in the heterosynaptic case appears to depend on postsynaptic mechanisms.     

Electrophysiological analysis of the dorsal hippocampal commissure projections to the entorhinal area

Synaptic effects evoked in the entorhinal area by dorsal hippocampal commissure (dorsal psalterium) projections were analysed in anesthetized adult guinea-pigs by means of a field potential analysis. Stimuli applied to the caudal part of the dorsal psalterium evoked a complex response in the dorsal third of the entorhinal area. The early part of the entorhinal response consisted of a slow wave interrupted by a spike potential. The electrophysiological characteristics and the laminar distribution of the slow wave and of the spike potential, together with the presence of time-locked unit activity, suggested that dorsal psalterium projections evoke monosynaptic excitatory postsynaptic potentials leading to cellular discharge in radially oriented neurons of layers II and III. The commissural fibers responsible for these effects originate in the contralateral presubiculum. The early part of the entorhinal response was followed by three waves in close temporal sequence. These waves were polysynaptically generated and associated with excitatory and inhibitory synaptic effects. Inhibition was demonstrated for the monosynaptically generated spike potential. Whether these effects were mediated by intracortical circuits and/or extrinsic projections cannot be stated from the present results. Causal relations were observed between the entorhinal monosynaptic response and that evoked by dorsal psalterium stimulation in the ipsilateral dentate gyrus, previously shown to be relayed by perforant path fibers. The results indicate that presubicular commissural projections to the entorhinal area monosynaptically activate neurons of the perforant pathway, whose discharge brings about activation of the ipsilateral dentate gyrus.     

Electrophysiological evidence for connections between septal neurones and the supraoptic nucleus of the hypothalamus of the rat

Summary In order to see whether septal neurones are connected to the hypothalamic neurones secreting vasopressin or oxytocin, neurones in different regions of the septum were recorded during electrical stimulation of the supraoptic nucleus. The position of the stimulating electrode within the latter was verified using lactating rats in which milk ejections could be induced by a train of electrical pulses applied to the nucleus. The responses of septal neurones to single pulse stimulation were then analysed by post-stimulus time histograms.In the septum ipsilateral to the site of stimulation, 42% of the neurones were antidromically invaded, 20% were orthodromically excited and 21% were inhibited following supraoptic stimulation. In the contralateral septum, 2% of the cells tested were antidromically invaded, 3% were excited and 16% inhibited. In the medial septum, 14% of the neurones were orthodromically excited, and 48% were inhibited.These results provide electrophysiological evidence for direct connections between septal neurones and the ipsilateral supraoptic nucleus of the hypothalamus, and give further support to the hypothesis of a septal influence on the activity of vasopressin- or oxytocin-releasing cells in the magnocellular system.Supported by grants I.N.S.E.R.M. CRL 79.5.372.6     

Frontolimbic lesions and social behavior in the rat

The social behavior of rats with chronic lesions to the medial frontal cortex, orbital frontal cortex, hippocampus, septum or amygdala was studied in four different situations: large open field, small open field, shock induced aggression and muricide. All lesions altered social behavior but the effects of the different lesions could be dissociated from one another. Lesions to the hippocampus dramatically reduced contact in open field tests and almost totally eliminated shock induced aggression. Lesions to the amygdala and septum also reduced the level of shock induced aggression but whereas amygdala lesions significantly decreased contact, septal lesions did not. Rats with lesions to the medial or orbital aspects of the prefrontal cortex differed from one another as the rats with orbital lesions exhibited consistently low contact scores and the rats with medial lesions did not, while both groups showed increased levels of shock induced aggression. None of the lesions significantly altered muricide.     

A topographical analysis of the origin of some efferent projections from the lateral hypothalamic area in the rat

Some projections from the lateral hypothalamic area in the rat have been investigated, using combinations of fluorescent tracers, injected into several different parts of the central nervous system. Projections appear to arise from loosely organized assemblies of neurons, called sets and from more densely packed assemblies, called clusters. The sets and clusters vary considerably in position and in distinctness of their borderlines. Even within extensive and vaguely defined sets, however, high concentrations of labeled neurons may be present at specified sites in the lateral hypothalamus. Such concentrations are observed in the transitional area of the zona incerta and the dorsal part of the lateral hypothalamus, and in the ventrolateral part of the hypothalamus, bordering the cerebral peduncle and the subthalamic nucleus, in both cases after injections into some "autonomic centers" in the brainstem, such as the parabrachial nuclei and the dorsal vagal complex. Sets and clusters may overlap considerably. Within the fields of overlap the number of double labeled neurons may vary from almost zero up to more than 50%, depending on the injection sites. The results show that different parts of the lateral hypothalamus in the rat have different efferent relationships. Combination of the results of the present study with known data concerning the afferent relationships, the cytoarchitecture and behavioral functions of the lateral hypothalamic area, suggests that different parts of this entity are involved in different regulatory and behavioral functions.     

Fimbria-fornix lesions and sexual-social behavior of the guinea pig

Male and female guinea pigs were given fimbria-fornix lesions and their sexual-social activity was compared to lesioned control animals. The lesions reduced the sexual and aggressive behavior of the male guinea pig. In the female, on the other hand, aggression towards the male increased during attempted copulation. Both experimental and control animals were given an open-field activity test. It was found that fimbria-fornix lesioned animals displayed less activity than did their control counterparts. These results were consistent with earlier findings and were related to previous observations of decorticate rats. It was suggested that the hippocampal output via the fimbria-fornix during sexual-social behavior plays a role in selecting motor patterns appropriate for the motivational state of the animal.     

Response pattern of cat hippocampal neurons to stimulation of the septal area during sleep and waking.

Unitary discharge was recorded extracellularly from cat dorsal hippocampus. The response pattern to electrical stimulation of the septum and regio limbica anterior was examined. There was a definite difference in the response pattern between the basket cell and the theta cell. Cluster analysis of high dimensional data was performed by means of Andrews' function plot. When compared with simple spike cells, complex spike cells showed a greater difference with one another with respect to the variation in the response pattern during different phases of sleep and wakefulness. The response patterns of complex spike cells showed a greater difference with one another during SS and PS than during A. It is suggested that the modulation of hippocampal activity during sleep and wakefulness is, as assessed from the reactivity to septal stimulation, operating more differentially at the output neurons than at its interneurons.     

Electrophysiological analysis of the vestibulospinal reflex pathway of rabbit. I. Classification of tract cells

Summary In anaesthetized rabbits, the medulla was surveyed with recording microelectrodes to identify different types of vestibulospinal tract neurones. Field potentials, unitary extracellular spikes and intracellular potentials were recorded during antidromic stimulation at C₁ and C₆ segments and during orthodromic stimulation through VIIIth nerve. The lateral and medial vestibulospinal tracts (LVST and MVST) were stimulated discriminately with the method developed in Appendix. On the basis of different axonal courses and conduction velocities, three major groups were distinguished for those cells which were activated monosynaptically by the primary vestibular afferents; 1. fast conducting LVST; 2. fast conducting MVST; and 3. slowly conducting MVST. Three other groups were discriminated for those cells which received only a polysynaptic or no action from primary vestibular afferents. These were; 4. fast conducting LVST; 5. slowly conducting LVST and 6. slowly conducting MVST. All of these six types of VST cells were represented within Deiters' nucleus. Only a relatively small number of MVST cells were found in the medial vestibular nucleus.     

The role of the epididymis in descensus testis and the topographical relationship between the testis and epididymis from the sixth month of pregnancy until immediately after birth

Summary The position of the testis, the relationship between the epididymis and the testis, as well as the development and regression of the gubernaculum were investigated in 18 testicles of children from the 26th week of pregnancy until a few weeks after birth. The most important role in descensus testiculorum is ascribed to the differentiation of the epididymis and the ductus deferens. It is androgen dependent. The testis descends in the processus vaginalis, being attached to its dorsal wall.