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T F Freund

Publications and source records attributed to T F Freund.

At least 91 records · Page 5Linked to original sources
PubMed1996

Interneurons containing calretinin are specialized to control other interneurons in the rat hippocampus.

Spine-free calretinin-immunoreactive (CR-IR) interneurons form a subpopulation of GABAergic cells in the rat hippocampus. A characteristic feature of these cells--located in all areas and layers--is the frequent dendro-dendritic and axo-dendritic contacts they form with each other. In this study we examined in detail the connectivity of these neurons by reconstructing their dendritic and axonal arbor and by identifying their postsynaptic targets. Radially running dendrites of CR-IR cells, located in different layers, intermingled into long braids. An average cell was in contact with dendrites of three to seven other CR-IR cells. Reconstruction of the dendritic trees from six consecutive sections demonstrated that at least 15 cells may participate in a dendro-dendritically connected cluster. Electron microscopical examination revealed that regularly spaced zonula adherentia connect the touching dendrites. The postsynaptic targets of CR-IR neurons have been examined using postembedding immunogold staining for GABA. CR-containing GABA-immunoreactive axons of local origin formed multiple symmetrical synaptic contacts (two to five) exclusively on GABAergic dendrites (CR-negative as well as CR-positive). Two to 10 CR-IR axons may converge onto a single CR-IR neuron, often from cells belonging to the same dendro-dendritically connected cluster. Using double immunocytochemistry, CR-IR cells were shown to heavily innervate calbindin D28k-containing interneurons and VIP-containing basket cells but avoided the parvalbumin-containing basket and axo-axonic cells. The unique connectivity of CR-IR cells may enable them to play a crucial role in the generation of synchronous, rhythmic hippocampal activity by controlling other interneurons terminating on different dendritic and somatic compartments of principal cells.

Animals↗
PubMed1996

Innervation of VIP-immunoreactive neurons by the ventroposteromedial thalamic nucleus in the barrel cortex of the rat.

We investigated the synaptic terminals of fibers originating in the ventroposteromedial thalamic nucleus (VPM) and projecting to the main input layers (IV/III) of the rat posteromedial barrel subfield. It was our aim to determine whether or not the subpopulation of vasoactive intestinal polypeptide (VIP)-immunoreactive neurons in these layers are directly innervated by the sensory thalamus. Anterograde tracing with Phaseolus vulgaris leucoagglutinin (PHA-L) and immunohistochemistry for VIP were combined for correlated light and electron microscopic examination. Columns of cortical tissue were well defined by barrel-like patches of PHA-L-labeled fibers and boutons in layers IV and III. Within these columns VIP-immunoreactive perikarya were located mainly in supragranular layers. Marked perikarya were also seen in infragranular layers, but their immunoreactivity was often weaker. Granular layer IV, which is the main terminal field for thalamic fibers, contained fewer VIP neurons than supragranular layers. In the light microscope, however, PHA-L-labeled fibers appeared to contact the somata or proximal dendrites of 60-86% of the layer IV VIP neurons . By contrast, only 18-35% of the VIP neurons in the supragranular layers, which receive a moderately dense projection from the VPM, appeared to be contacted. PHA-L-labeled boutons were seen close to 13-25% of infragranular VIP-positive cells. Electron microscopy showed that thalamic fibers formed at most four asymmetric synapses on a single layer IV, VIP-positive neuron. Although the proportion of VIP-positive neurons with labeled synapses was lower in supragranular layers, most of them shared multiple asymmetric synapses with labeled thalamic fibers. Up to six labeled synapses were seen on individual VIP neurons in layer III. We conclude that subpopulations of VIP-immunoreactive neurons, located in layers IV, III, and II are directly innervated by the VPM. These neurons may be involved in the initial stages of cortical processing of sensory information from the large, mystacial vibrissae. Since VIP is known to be colocalized with the inhibitory transmitter GABA, it is likely that VIP neurons participate in the shaping of the receptive fields in the barrel cortex.

Afferent Pathways↗
PubMed1996

Pyramidal cell dendrites are the primary targets of calbindin D28k-immunoreactive interneurons in the hippocampus.

The axonal arborization and postsynaptic targets of calbindin D28k (CB)-immunoreactive nonprincipal neurons have been studied in the rat dorsal hippocampus. Two types of neurons were distinguished on the basis of soma location, the characteristics of the dendritic free, and the axon arborisation pattern. Type I cells were located in stratum radiatum of the CA1 and CA3 regions and occasionally in strata pyramidale and oriens. These cells had multipolar or bitufted dendritic trees primarily located in stratum radiatum. Their axons could be followed for a considerable distance, arborised within stratum radiatum, and were covered with regularly spaced small boutons. As demonstrated with postembedding immunogold staining, their axon terminals were gamma-aminobutyric acid (GABA) immunoreactive, and formed symmetrical synapses predominantly on proximal and distal dendrites of pyramidal cells (28% and 58%, respectively), and occasionally on spines (9%) or on GABA-positive dendrites (5%). Type II cells were found exclusively in stratum oriens of the CA1 and CA3 regions and possessed large, fusiform cell bodies and long, horizontally oriented dendrites. Their axon initial segments turned towards the alveus and disappeared in a myelin sheet, which was often possible to follow into the white matter. We conclude that type I CB-immunoreactive cells are likely to represent a major source of inhibitory synapses in the dendritic region of pyramidal cells, which are responsible for the control of dendritic electrogenesis. The distribution of local collaterals of type II cells-if they have any-remains unknown, but their main axon is likely to project to the medial septum.

Animals↗
PubMed1996

Recurrent axon collaterals of corticothalamic projection neurons in rat primary somatosensory cortex contribute to excitatory and inhibitory feedback-loops.

Intrinsic circuitry within the primary somatosensory cortex of the rat was examined in a combined light and electron microscope study. Corticothalamic projection neurons were retrogradely labeled by applying Phaseolus vulgaris leucoagglutinin (PHA-L) into the ventro-posteromedial thalamic nucleus (VPM). Most labeled neurons were pyramidal cells of layer VI. Postsynaptic targets of recurrent axon collaterals originating from these neurons were assessed in layers IV and V. Single labeled cells, complete with recurrent collaterals, could be isolated in "barrels" in which no anterograde transport had taken place. These findings were confirmed by first eliminating thalamocortical projections from the VPM with kainic acid and then applying PHA-L into the same nucleus. This procedure led to selective retrograde accumulation of tracer in layer VI pyramidal cells. Reconstructed portions of labeled axonal trees reached layer IV, bringing numerous boutons to layers IV, V and VI. The boutons had characteristic drumstick-like shapes. In order to identify postsynaptic targets, 4 sections of axons stemming from 3 neurons were reembedded and serially sectioned for electron microscopy. The ultrastructure of 72 asymmetric synapses, all belonging to identified collaterals, was analysed. Of the 72 terminals, 44 (59.5%) ended on dendritic spines and 30 on shafts of dendrites (40.5%). Perikarya were not among the targets. In a subset of the sample, the nature of the target neurons was examined by postembedding immunohistochemistry for gamma-amino butyric acid (GABA) after staining for PHA-L. A total of 42 labeled terminals was found in layers IV and V; 23 (55%) were located on GABA-negative spines and 19 (45%) on dendritic shafts. Only 6 (32%) of the shafts were GABA-positive. The remaining ones were either clearly GABA-negative, or labeled only at background levels (n = 13; 68%). The results show that most synapses of corticothalamic projection neurons found in layers IV and V terminate on spines and shafts of GABA-negative dendrites. This finding suggests that such recurrent collaterals are involved in both excitatory and inhibitory feedback mechanisms.

Animals↗
PubMed1996

Regeneration of the GABAergic septohippocampal projection in vitro.

The formation of the GABAergic septohippocampal projection was studied in vitro. Slice cultures of the septal complex from young postnatal rats were prepared and co-cultivated with hippocampal slices for up to four weeks. Then, the anterogradely transported tracer Phaseolus vulgaris leucoagglutinin was injected into the septal culture and the labeled fibers were traced into the hippocampal culture. Some fibers were identified as originating from GABAergic septal cells by double-labeling with an antiserum against GABA using the postembedding immunogold procedure. Our results showed that double-labeled terminals of GABAergic septohippocampal neurons established symmetric synapses exclusively with GABA-positive dendrites in one out of five co-cultures, but also contacted numerous GABA-negative structures in the remaining four co-cultures. These findings, together with light microscopic data from sections double-stained for Phaseolus and parvalbumin, indicate that the high target selectivity of the GABAergic septohippocampal pathway for GABAergic interneurons in vivo is lost in most cases, at least under the present in vitro conditions. It is hypothesized that this may be due to an immaturity of the connection, the lack of axon-guiding factors or an expansion of the septohippocampal GABAergic fibers in the absence of many extrinsic afferents, including GABAergic fibers. The simultaneous occurrence of anterogradely labeled, but GABA-negative, septohippocampal terminals in the hippocampal target culture also suggests that the septohippocampal cholinergic projection developed in vitro, as was shown before in other studies. Since most septohippocampal neurons have to be axotomized for culture preparation, the present results indicate that GABAergic septohippocampal neurons from young postnatal rats survive axotomy and are capable of regenerating a septohippocampal projection, including the formation of characteristic GABAergic synapses on co-cultured hippocampal neurons. However, the characteristic target selectivity is rarely preserved.

Animals↗
PubMed1996

Different populations of vasoactive intestinal polypeptide-immunoreactive interneurons are specialized to control pyramidal cells or interneurons in the hippocampus.

The postsynaptic targets of three vasoactive intestinal polypeptide-containing GABAergic interneuron types were examined in the rat hippocampus. Two of them showed remarkable target selectivity for other GABAergic neurons, while the third contacted the somata and proximal dendrites of pyramidal cells. Vasoactive intestinal polypeptide-positive interneurons innervating the stratum oriens/alveus border in the CA1 region were shown to establish multiple contacts with horizontal GABAergic interneurons immunoreactive for type 1 metabotropic glutamate receptor. Similarly, identified axons of vasoactive intestinal polypeptide-positive interneurons projecting to stratum radiatum were found to establish symmetrical synapses largely on GABAergic dendrites. The majority of these postsynaptic GABAergic neurons were shown to contain calbindin or vasoactive intestinal polypeptide. In contrast to the first two vasoactive intestinal polypeptide-containing cell populations, vasoactive intestinal polypeptide-positive interneurons arborizing in stratum pyramidale formed baskets around pyramidal cells. These results revealed a new element in cortical microcircuits, interneurons which are specialized to innervate other GABAergic interneurons. The role of this new component may be the synchronization of dendritic inhibition, or an input-specific disinhibition of pyramidal cells in various dendritic domains. In contrast, vasoactive intestinal polypeptide-containing basket cells are likely to be involved in perisomatic inhibition of pyramidal neurons, and represents a new basket cell type different from that containing parvalbumin.

Animals↗
PubMed1996

Correlated morphological and neurochemical features identify different subsets of vasoactive intestinal polypeptide-immunoreactive interneurons in rat hippocampus.

Vasoactive intestinal polypeptide-immunoreactive interneurons have been classified according to their axonal and dendritic patterns and neurochemical features in the hippocampus of the rat. A correlation of these characteristics unravelled three distinct types of vasoactive intestinal polypeptide-containing cells. Interneurons forming a dense axonal plexus at the border of stratum oriens and alveus always contain the calcium binding protein, calretinin, but lack the neuropeptide cholecystokinin. The axon of another type of vasoactive intestinal polypeptide-positive interneuron surrounds pyramidal cell bodies in a basket-like manner, and co-localizes cholecystokinin but not calretinin. Vasoactive intestinal polypeptide-containing cells projecting to stratum radiatum form two subsets distinguished by dendritic morphology. Those with dendrites restricted to stratum lacunosum-molecular lack both calretinin and cholecystokinin, whereas the other subtype with dendrites spanning all layers contains calretinin in 40% of the cases and occasionally also cholecystokin. GABA was shown to be present, and the calcium binding proteins calbindin D-28k and parvalbumin absent from all three types of vasoactive intestinal polypeptide-positive interneurons. The specific dendritic and axonal arbours imply different input and output properties for the three interneuron types. The correlation of these features with the content of neurochemical markers strongly suggests that they are specialized for distinct inhibitory functions in the hippocampal network.

Animals↗
PubMed1996

Differences between somatic and dendritic inhibition in the hippocampus.

Hippocampal synaptic inhibition is mediated by distinct groups of inhibitory cells. Some contact pyramidal cells perisomatically, while others terminate exclusively on their dendrites. We examined perisomatic and dendritic inhibition by recording from CA3 inhibitory and pyramidal cells and injecting biocytin to visualize both cells in light and electron microscopy. Single perisomatic inhibitory cells made 2-6 terminals clustered around the soma and proximal pyramidal cell processes. Dendritic cells established 5-17 terminals, usually on different dendrites of a pyramidal cells. Perisomatic terminals were larger than those facing dendritic membrane. Perisomatic inhibitory cells initiated the majority of simultaneous IPSPs seen in nearby pyramidal cells. Single IPSPs initiated by perisomatic sodium-dependent action potentials. Activation of inhibitory fibers terminating on dendrites could suppress calcium-dependent spikes. Thus, distinct inhibitory cells may differentially control dendritic electrogenesis and axonal output of hippocampal pyramidal cells.

Action Potentials↗
PubMed1996

Distribution of GABAergic elements postsynaptic to ventroposteromedial thalamic projections in layer IV of rat barrel cortex.

The spatial synaptic pattern formed by boutons, originating in the ventroposteromedial thalamic nucleus, with GABAergic neurons in the rat barrel cortex was mapped. The aim was to shed light on the structural basis by which inhibitory circuits may be activated at the first stage of cortical information processing. The thalamic afferent projection was labelled by anterograde transport of Phaseolus vulgaris leucoagglutinin (PHA-L), whereas the GABAergic targets in layer IV of the rat barrel cortex was visualized by postembedding GABA immunogold-labelling or by pre-embedding parvalbumin immunocytochemistry. In the first set of experiments, we mapped barrels, contained in single ultrathin sections, by means of a computer-controlled electron microscope stage in their entire layer IV representation. From a total of 1199 asymmetric PHA-L-labelled synapses, only 98 were on GABAergic elements, mainly on dendritic shafts. This corresponded to 8.2% of all synapses counted. These synapses on GABAergic targets were essentially homogeneously distributed without a reliable relationship to barrel subdivisions, i.e., hollow versus wall; or layer IVa versus layer IVb. In the second part of the study, we demonstrated that parvalbumin-containing neurons represent the major GABAergic cell type targetted by thalamic afferents in layer IV of the barrel cortex, since all parvalbumin-positive cells investigated received multiple synaptic contacts (up to eight synapses per neuron) from the ventroposteromedial thalamic nucleus. These results imply that interneurons responsible for perisomatic inhibition (basket and chandelier cells known to contain parvalbumin) are likely to be strongly excited by thalamic afferents, despite the relatively low proportion of thalamic synapses on GABAergic elements compared to spines of principal cells, and participate in the early stages of cortical sensory information processing.

Animals↗
PubMed1996

Target selectivity and neurochemical characteristics of VIP-immunoreactive interneurons in the rat dentate gyrus.

Vasoactive intestinal polypeptide (VIP) has been shown to be present in a morphologically heterogeneous subpopulation of interneurons in the dentate gyrus, but the relationship between their input and output characteristics and neurochemical features has not been established. Three types of VIP-immunoreactive cells have been identified on the basis of these criteria: (i) cells forming a dense axonal plexus in the hilus have always coexisted with the calcium binding protein calretinin (CR), but never with the neuropeptide cholecystokinin (CCK). The postsynaptic targets of these VIP-positive cells were neurons visualized by immunostaining for substance P receptor, which is known to label different hilar non-principal cells. (ii) VIP-immunoreactive basket cells, innervating predominantly the somata and proximal dendrites of granule cells, were found in the striatum moleculare and stratum granulosum. They contained CCK, but not CR. (iii) Cells projecting to the stratum moleculare were found to have dendrites and axons restricted to this layer. In 75% of these cells VIP coexisted with CR but not with CCK, and they established multiple contacts largely with non-principal cells. GABA was shown to be present but the calcium-binding proteins calbindin D28K and parvalbumin were absent in all three types of VIP-containing interneuron. On the basis of these observations we conclude that three different types of VIP-positive neuron are present in this area, and are likely to subserve different inhibitory functions, cells with a hilar projection as well as those projecting to the stratum moleculare may synchronize the activity of hilar and other interneurons, or disinhibit granule cells by specific interneuron-to-interneuron connections. In contrast, basket cells control the activity of granule cells directly, via perisomatic inhibition.

Animals↗
PubMed1996

Topographic distribution of dorsal and median raphe neurons with hippocampal, septal and dual projection.

Double retrograde neuronal tracing techniques with used to determine the distribution and degree of overlap between cell population in the raphe nuclei with a projection to the medial septum and/or the hippocampus. The results of the present study showed that numerous median raphe neurons that innervated the hippocampus, sent a collateral projection to the medial septum. In contrast to that, a spatially segregated population of serotonergic neurons located caudally in the dorsal raphe nucleus projected only to the medial septum but not to the hippocampus. Our results show that the medial septum receives dual serotonergic innervation from the midbrain raphe nuclei. One projection arises from axon collaterals of neurons that contribute to the median raphe-hippocampal pathway as well. The second serotonergic input originates from the caudal part of the dorsal raphe nucleus, and innervates the medial septum, but ignores the hippocampus. These findings suggest that, in addition to the well-known serotonergic effect of the median raphe on hippocampal electrical activity, theta rhythm in the hippocampus may also be modulated by the dorsal raphe nucleus via the medial septum.

Animals↗
PubMed1995

Synaptic input of horizontal interneurons in stratum oriens of the hippocampal CA1 subfield: structural basis of feed-back activation.

The synaptic input of interneurons with horizontal dendrites in stratum oriens of the CA1 region was investigated, with particular attention to the portion of synapses originating from local pyramidal cells. Most of these GABAergic interneurons are known to contain somatostatin, and terminate on pyramidal dendrites in conjunction with entorhinal afferents in stratum lacunosum-moleculare. A smaller number of horizontal cells in this layer are immunoreactive for calbindin, and project to the medial septum. Selective ischaemic degeneration was used to label local axon collaterals of CA1 pyramidal cells, and immunostaining for mGluR1 or calbindin to visualise somatostatin- and calbindin-containing horizontal interneurons, respectively, at the stratum oriens-alveus border. The number of degenerating and intact synaptic boutons was counted on mGluR1- as well as on calbindin-positive dendrites and somata, whereas in another group of animals the proportion of GABA-immunoreactive synapses was estimated on calbindin-positive dendrites. On average, > 60% of the total presynaptic elements of both cell types were degenerating, i.e. originated from CA1 pyramidal cells, whereas GABA-positive boutons, which are known to survive ischaemia, are likely to account for a large proportion of non-degenerating boutons. Thus the vast majority of presumed excitatory synapses on somatostatin- and calbindin-containing horizontal neurons derives from local collaterals of CA1 pyramidal cells. The remaining GABA-negative synapses surviving ischaemia may also originate from CA1 pyramidal cells, e.g. from those in the ventral hippocampus, which are rarely damaged by global forebrain ischaemia. Alternative sources may include subcortical afferents known to innervate interneurons, or ipsi- and contralateral CA3 pyramidal cells, which, according to the present results, may account only for a negligible number of synapses on these interneurons types. We conclude that somatostatin-containing neurons at the oriens-alveus border of CA1, which are likely to mediate an inhibitory control of the efficacy and/or plasticity of entorhinal synapses on pyramidal cell dendrites, are driven primarily in a feed-back manner. The source of afferent excitation for calbindin-containing horizontal neurons in this region is very similar, suggesting that the GABAergic hippocamposeptal feed-back is also activated by local pyramidal cell collaterals.

Afferent Pathways↗
PubMed1995

Delayed cell death in the contralateral hippocampus following kainate injection into the CA3 subfield.

A model of epileptic cell death has been developed employing unilateral injections of kainic acid, a glutamate agonist, into the CA3 subfield of the hippocampus. The contralateral hippocampus, where neuronal damage is induced by hyperactivity in afferent pathways, served as the model structure. The pattern of cell death in this model was shown earlier to correspond to the vulnerable regions in human temporal lobe epilepsy. In the present time-course study we demonstrated that the different subpopulations of vulnerable cells in the contralateral hippocampus of the rat degenerate at different times following kainate injection. Spiny calretinin-containing cells in the hilus and CA3 stratum lucidum disappear at 12-24 h, other types of hilar neurons and CA3c pyramidal cells show shrinkage and argyrophilia at two days, whereas CA1 pyramidal cells degenerate at three days postinjection. The majority of cells destined to die showed a transient expression of the heatshock protein 72, approximately one day (for hilar-CA3c) or two days (for CA1) before degeneration. Parvalbumin-immunoreactivity transiently disappeared from the soma and dendrites of interneurons between the first and the fourth day. The results suggest that seizure-induced cell death is delayed, therefore acute oedema, even if it occurs, is insufficient to kill neurons. The only exception is the population of calretinin-containing interneurons degenerating at 12-24 h. The further one day delay between hilar-CA3c and CA1 cell death is likely to be due to differences in the relative density of glutamate receptor types (kainate versus NMDA) and the source of afferent input of these subfields. Thus, simple pharmacotherapy targeting only one of the excitotoxic mechanisms (i.e. acute oedema of calretinin cells versus delayed death of hilar-CA3c and CA1 cells at different time points) is likely to fail.

Animals↗
PubMed1995

Development of the raphe-hippocampal projection in vitro.

In this study we examined whether the serotonergic raphe-hippocampal projection preserves its characteristic target selectivity for GABAergic interneurons when developing in vitro, in organotypic cultures. Hippocampal slices from one- to three-day-old rats were co-cultured with slices derived from the raphe nuclei of the same animals. After several weeks of in vitro incubation, a large number of raphe fibres--visualized by immunostaining for serotonin--were found to innervate the hippocampal tissue. In our random sample of over 250 serially sectioned boutons--52 of which were completely reconstructed from serial sections--only two were found to form conventional synapses in the electron microscope, and contacted dendritic spines. These results demonstrate that raphe-hippocampal serotonergic afferent are unable to form synaptic contacts with their normal targets in vitro, if explanted one to three days postnatally. Neurons in the afferent and/or target area may have passed a critical age when selective synaptic contacts can be formed, or unknown chemical or electrical signals may be missing under these conditions, which should serve to guide subcortical afferents to their synaptic target elements.

Animals↗
PubMed1994

Differential effects of serotonin and raphe grafts in the hippocampus and hypothalamus: a combined behavioural and anatomical study in the rat.

Combined with a partial cholinergic deficiency, serotonergic lesions induce severe spatial learning deficits. Serotonergic lesions, however, have additional effects, such as reduced body weight and disruption of thermoregulation, which may be the cause of the observed learning deficits. Restoration of the serotonergic innervation of the hippocampus by raphe grafts reduces these learning deficits. The effects of the grafts may result from a direct support of spatial learning but may also be an indirect result of preventing some of the other effects of serotonergic lesions. In the present study we used raphe grafts to examine the selectivity and specificity of the effects of serotonergic lesions in the rat, and used the behavioural effects as an indication of successful transplantation in order to examine the fine details of such grafts. Raphe grafts in the hippocampus did not prevent the effects of the lesions on body weight, thermoregulation and exploratory behaviour but did minimize the effects of the lesions on spatial learning. In contrast, raphe grafts in the hypothalamus reduced the effects of the lesions on thermoregulation but failed to support learning. The grafted fibres showed termination specificity with the interneurons, which is typical of the serotonergic innervation of the normal hippocampus. The results indicate that the serotonergic innervation of the hippocampus functions locally to support spatial learning. This role of serotonin is independent of its involvement in modulation of body weight, thermoregulation or exploratory behaviour. The results confirm that the modes of serotonergic action in the hippocampus include the selective innervation of specific interneuron subpopulations.

5,7-Dihydroxytryptamine↗
PubMed1994

Serotoninergic innervation of nonprincipal cells in the cerebral cortex of the lizard Podarcis hispanica.

The mechanism of serotoninergic transmission in the neo- and archicortex of mammals is complex, including both synaptic and nonsynaptic components, direct actions on principal cells, and indirect effects mediated by GABAergic interneurons. Here we studied the termination pattern and synaptic organization of the serotoninergic afferents in the cerebral cortex of the lizard, Podarcis hispanica, which is considered to correspond in part to the mammalian hippocampal formation, with the aim of unraveling basic, phylogenetically preserved rules in the connectivity of this pathway. We demonstrate that serotoninergic afferents, visualized by immunostaining for serotonin itself, establish multiple synaptic contacts with different subpopulations of nonprincipal cells containing parvalbumin, neuropeptide Y, and opioid peptides. The former two subpopulations contain GABA, whereas the opioid-immunoreactive neurons are most likely GABA-negative cells. Evidence is provided at the electron microscopic level that serotonin-immunoreactive varicosities establish conventional asymmetric synaptic contacts with their nonprincipal targets, but nonsynaptic varicosities also exist. We conclude that, similarly to mammals, a selective synaptic innervation of nonprincipal, possibly inhibitory, neurons is among the mechanisms of serotoninergic modulation of cerebral cortical activity in the lizard.

Animals↗
PubMed1994

Principal cells are the postsynaptic targets of supramammillary afferents in the hippocampus of the rat.

Neurons of the supramammillary nucleus are known to fire phase-locked to hippocampal theta rhythm. Stimulation of this area induces theta activity in the hippocampus via the medial septum and facilitates perforant pathway stimulation-evoked population spikes in the dentate gyrus even if the medial septum is inactivated. This latter effect was suggested to be due to a direct inhibitory input from the supramammilary nucleus to hippocampal nonpyramidal cells resulting in disinhibition. In the present study, using anterograde tracing with Phaseolus vulgaris leucoagglutinin, we aimed to identify the types of neurons innervated by the supramammillary projection in the dentate gyrus and Ammons horn, with particular attention to the presumed postsynaptic inhibitory neurons, which may mediate the proposed disinhibitory action. Double-immunostaining for the tracer and different neuropeptides (somatostatin, cholecystokinin, neuropeptide Y) or calcium binding proteins (calretinin, parvalbumin, calbindin D28K) present in different subpopulations of interneurons revealed no multiple contacts between supramammillary afferents and labeled inhibitory cells at the light microscopic level. Furthermore, postembedding immunostaining of electron microscopic sections for GABA demonstrated that none of the 68 PHAL-labeled supramammillary boutons examined and none of their postsynaptic targets were immunoreactive for the inhibitory neurotransmitter. We conclude, therefore, that most if not all postsynaptic targets of the supramammillary projection are principal cells both in the dentate gyrus and in the CA2-CA3a subfields. This suggests that a mechanism other than disinhibition is responsible for the facilitatory effect of this pathway on hippocampal evoked activity.

Afferent Pathways↗