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B Heimrich

Publications and source records attributed to B Heimrich.

At least 37 records · Page 2Linked to original sources
PubMed1997

Sprouting in the hippocampus is layer-specific.

Partial removal of layer-specific afferents of the hippocampus is said to induce sprouting of intact fibers from neighboring layers that invade the zone of the degenerating axons. However, recent in vivo and in vitro studies using sensitive anterograde tracers have failed to demonstrate sprouting across laminar boundaries. Sprouting does occur; but, it mainly involves unlesioned fiber systems terminating within the layer of fiber degeneration in addition to the degenerating afferents. These findings point to rigid laminar cues attracting certain fiber systems while repelling others in normal development and after partial deafferentation.

Animals↗
PubMed1997

Histaminergic system in co-cultures of hippocampus and posterior hypothalamus: a morphological and electrophysiological study in the rat.

Neurons of the tuberomammillary nucleus in the posterior hypothalamus diffusely project to most parts of the central nervous system, where their main transmitter, histamine, modulates the excitability of the target neurons. The development of a histaminergic hypothalamo-hippocampal pathway and its function were studied in organotypic co-cultures. Immunocytochemistry for histidine decarboxylase, the specific synthesizing enzyme, stained clusters of neurons in the hypothalamic tuberomammillary area. Immunolabelled varicose processes innervated the co-cultured hippocampus and established a few synaptic contacts on dendrites. Cultured tuberomammillary neurons displayed their typical membrane properties and were spontaneously active. In hippocampal pyramidal cells of the CA3 region the long-lasting afterhyperpolarization was reduced by histamine or impromidine and increased by the H2 antagonist cimetidine, but not by the H1 antagonist mepyramine. The membrane potential was depolarized in presence of an H2 agonist and hyperpolarized by an H2 antagonist. In single hippocampal cultures histamine antagonists did not affect afterhyperpolarization and membrane potential. Histaminergic neurons retain their main morphological and physiological characteristics in slice cultures and establish a functional connection with co-cultured target cells.

Animals↗
PubMed1996

Differential survival of Cajal-Retzius cells in organotypic cultures of hippocampus and neocortex.

Cajal-Retzius (CR) cells are transient, pioneer neurons of layer I of the cortex that are believed to play essential roles in corticogenesis, e.g., in neuronal migration and synaptogenesis. Here we have used calretinin immunostaining to study the characteristics, survival, and fate of CR cells in single organotypic slice cultures of mouse neocortex and hippocampus deprived of their extrinsic afferents. In neocortical explants, CR cells were observed after 1-3 d in vitro (DIV), but they disappeared after 5-7 DIV, which is similar to their time of degeneration in vivo. The disappearance of CR cells in neocortical slices was prevented by incubation with tetrodotoxin and the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3,-dione but not by 2-amino-5-phosphonopentanoic acid, suggesting that neuronal activity and non-NMDA glutamate receptors may trigger CR cell death in the neocortex. In contrast to the situation in vivo, in which many hippocampal CR cells disappear at approximately the third postnatal week, CR cells survived in single hippocampal cultures after long incubation times (31 DIV), with their morphology essentially unaltered. In contrast, fewer CR cells were found when hippocampal slices were cocultured with explants from the entorhinal cortex. Because CR cells are transient synaptic targets for entorhinohippocampal afferents, these findings suggest a role for entorhinal afferents in the degeneration of CR cells in the hippocampus. In conclusion, this study shows different survival properties of CR cells in organotypic slice cultures of hippocampus and neocortex, and it suggests that different mechanisms are involved in the regulation of the process of naturally occurring CR cell death in the two cortical regions.

Afferent Pathways↗
PubMed1996

Expression of the proenkephalin A gene in organotypic cultures of neocortex from newborn rats.

In rats, the proenkephalin A gene is expressed in proliferating cells of the neuroepithelial zone which later give rise to neocortical neurones and glial cells. Therefore, organotypic cultures of neocortex of newborn rats were used in the present study to examine whether neurones as well as glial cells expressed the gene. The slices were prepared at birth and kept in culture for 7-13 days. Proenkephalin mRNA was visualised by in situ hybridisation, while immunocytochemical staining for MAP-2 and GFAP was used to identify neurones and astroglial cells, respectively. In the analysed slices, only neurones contained proenkephalin mRNA. Activation of protein kinase C with tetradecanoylphorbol acetate (1 mumol/l) caused a strong increase in the number of neurones expressing proenkephalin mRNA. Our results indicate that a large number of neurones is able to express the proenkephalin gene under these conditions. However, only a few of them have a basal expression which is strong enough to be detected with in situ hybridisation.

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

Survival, regeneration and sprouting of central neurons: the rat septohippocampal projection as a model.

The septohippocampal projection was used to study the survival following axotomy, axonal regeneration, and sprouting of a defined group of central neurons. Septohippocampal projection neurons in adult rats were axotomized by bilateral lesions of the fimbria-fornix. Using prelabeling prior to axotomy, intracellular staining, electron microscopy, and immunocytochemical and in situ hybridization techniques, we were able to demonstrate that the majority of septohippocampal neurons survived after axotomy. At least in young postnatal rats, these axotomized neurons have the capacity to regenerate an axonal process that reinnervates its appropriate target tissue, the hippocampus. We demonstrated this by axotomizing young septohippocampal neurons and co-culturing them with sections of hippocampus. Septohippocampal neurons appear to retain their capacity for axonal growth in adulthood, since they are able to sprout within hippocampal layers partially denervated by removing entorhinal afferents. In this paradigm the terminals of septohippocampal neurons themselves were not lesioned. Our results point to a previously underestimated capacity of septohippocampal neurons for survival following axotomy, regeneration, and sprouting.

Acetylcholinesterase↗
PubMed1996

Differentiation of Purkinje cells in cerebellar slice cultures: an immunocytochemical and Golgi EM study.

In the rat central nervous system, the cerebellar cortex has a stereotypical cytoarchitecture and a characteristic connectivity pattern, both mainly formed post-natally. Organotypic cultures of immature cerebellar tissue were used to study the formation of the cerebellar lamination and the differentiation of Purkinje cells in the absence of their extracerebellar afferents. The lamination was retained in the majority of the cerebellar cultures and most Purkinje cells were aligned. Axonal profiles of Purkinje cells, immunolabelled for UCHT1 or anti-calbindin D-28 k, followed pathways similar to those in vivo cerebellum. The dendrites were orientated towards the superficial layer except of those neurons which were ectopically positioned. Unlike in vivo, the dendritic arborization of Golgi-impregnated/gold-toned or immunostained Purkinje cells was reduced and the dendritic spines were often elongated. Somatic spines, a morphological feature of immature Purkinje cells persisted even after 4 weeks in culture. We conclude that the Purkinje cells in organotypic cultures send their axon to the correct target region independent of their local position. In contrast, the dendritic orientation and differentiation is influenced by the cellular environment and by specific synaptic interaction.

Animals↗
PubMed1995

Restoration of mossy fiber projection in slice co-cultures of dislocated dentate gyrus and degranulated hippocampus.

Regional specificity of the mossy fiber projection is a well described feature of hippocampal intrinsic connectivity. Possible mechanisms involved in the formation of this specific projection include attraction molecules localized in the target area or repulsive cues preventing from ingrowth in non-target areas. To test this hypothesis, using organotypic co-cultures of dentate gyrus and irradiated degranulated hippocampal slices, we have disrupted the pathway normally taken by mossy fibers. The dentate gyrus explant was ectopically placed facing the alveus/stratum oriens of the irradiated hippocampal slice forcing the mossy fibers to cross the stratum oriens to reach their target area. Extensive plexuses of labeled mossy fibers were observed in the hilus and adjacent pyramidal cell layer of non-irradiated dentate gyrus explants. A few mossy fibers crossed the border between the co-cultures and reached their specific termination area in the irradiated hippocampus where they formed characteristic multiple synaptic contacts on their target cells. In addition to mossy fibers, numerous thin and varicose non-mossy fibers invade all parts of the co-cultured hippocampus establishing symmetric synapses. From these data we assume that mossy fiber axons emerging from dislocated non-irradiated dentate gyrus explants find their normal termination zone in the co-cultured degranulated hippocampal slice even if they are forced to run an unusual pathway. These results support the idea that an attraction signal arising from the target area is involved in the formation of this specific projection.

Animals↗
PubMed1995

Lamina-specific synaptic connections of hippocampal neurons in vitro.

By using slice cultures as a model, we demonstrate here that different target selectivities exist among the various afferent fibers to the hippocampus. As in intact animals, septohippocampal cholinergic fibers, provided by a slice culture of septum, innervate a co-cultured slice of hippocampus diffusely, that is, without forming distinct layers of termination. As in vivo, the septal cholinergic fibers establish synapses with a variety of target cells. Conversely, fibers from an entorhinal slice co-cultured to a hippocampal slice display their normal laminar specificity. They preferentially terminate in the outer molecular layer of the fascia dentata, thereby selectively contacting peripheral dendrites of the granule cells. This preferential termination on peripheral dendritic segments is remarkable, since these fibers do not have to compete with commissural fibers, hypothalamic fibers, and septal afferents for dendritic space under these culture conditions. Moreover, in triplet cultures in which first two hippocampal slices were co-cultured and then, with a delay of 5 days, an entorhinal slice was added, the fibers from the entorhinal slice and those from the hippocampal culture terminated in their appropriate layers in the hippocampal target culture. However, in this approach the normal sequence of ingrowth of these two afferents was reversed. In normal ontogenetic development, entorhinal afferents arrive in the hippocampus before the commissural fibers. The results show that there are different degrees of target selectivity of hippocampal afferents and that the characteristic lamination of certain afferent fibers in the hippocampus is not determined by their sequential ingrowth during development.

Afferent Pathways↗
PubMed1995

Axonal regeneration of identified septohippocampal projection neurons in vitro.

Previous studies have shown that the septohippocampal projection develops in vitro. It remained unclear, however, whether this in vitro projection resulted from axonal regeneration of septal neurons transected by the culture preparation, or from a priori outgrowth of undamaged cells that had not yet sent an axonal process to the hippocampus at the time of tissue preparation. Here we have prepared slice cultures of identified septohippocampal projection neurons that were retrogradely labeled by hippocampal tracer injection prior to slice preparation. We demonstrate that these identified projection neurons survive axotomy caused by culture preparation, are able to regrow an axonal process in vitro, and innervate a co-cultured hippocampal slice.

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↗
PubMed1995

Understanding the cortex through the hippocampus: lamina-specific connections of the rat hippocampal neurons.

A characteristic feature of hippocampal organisation is the lamina-specific termination of afferent fibres. The factors determining this characteristic fibre segregation are not known. By using slice cultures as a model, we have recently demonstrated that the laminated termination of hippocampal afferents does not result from the temporal sequence of ingrowth of the various afferent fibre systems during development. Moreover, the lack of extrinsic afferents in culture does not induce a substantial translaminar sprouting of the remaining (intrinsic) fibres or of a defined afferent system supplied by a coculture. These results contrast with previous reports on an expansion of intact fibre systems in the hippocampus in response to partial deafferentation, for instance by removal of entorhinal afferents. We therefore studied the sprouting of commissural fibres following an entorhinal lesion directly by labelling commissural axons in vivo with the anterogradely transported tracer Phaseolus vulgaris leucoagglutinin. Sprouting of commissural fibres following an entorhinal lesion was observed but, in accordance with our in vitro observations, this growth of commissural terminals took place within the appropriate termination zones of commissural fibres. These results point to a rigid laminar specificity of hippocampal afferents that is preserved after partial deafferentiation.

Animals↗
PubMed1994

Dendritic development of dentate granule cells in the absence of their specific extrinsic afferents.

Dendrites and spines are postsynaptic structures that develop in association with presynaptic fibers. Recent studies have shown that granule cells of the fascia dentata survive in slice cultures and differentiate in a manner known from in situ studies. However, all extrinsic afferent fibers are absent under culture conditions. In the present study, we study whether dendrites and spines of granule cells in slice cultures differentiate normally, although they are not contacted by their normal layer-specific afferents. Slices of hippocampus were prepared from rat pups at the day of birth. After 5, 10, 15, and 20 days of incubation, granule cells in these cultures were Golgi impregnated. For comparison, perfusion-fixed hippocampal sections of 5-, 10-, 15-, and 20-day-old rats were impregnated the same way. Our results show that the total density of spines on granule cell dendrites in culture increased as in perfusion-fixed animals. However, after 20 days of incubation, the absolute number of dendritic spines on cultured neurons was reduced because of a reduction of peripheral dendrites. This reduction was accompanied by an increase in the number of stem dendrites originating from the perikaryon. The density of spines on these proximal dendrites was larger in cultured granule cells than in controls. Our results suggest that the lack of major extrinsic (entorhinal) afferents that normally terminate on peripheral granule cell dendrites causes retraction of these dendrites. At the same time, there is growth of proximal dendritic portions. Proximal dendrites are targets of associational fibers, which are known to sprout under these culture conditions.

Animals↗
PubMed1993

Formation of layer-specific fiber projections to the hippocampus in vitro.

The factors determining the layer-specific termination of hippocampal afferents are not known. Previous studies have suggested that the laminated termination of afferent fiber systems is caused by their sequential ingrowth during development. Here we have tested this temporal hypothesis of fiber segregation by an in vitro confrontation system in which the sequential arrival of entorhinal and commissural fibers was reversed. However, despite the temporal reversal of ingrowth, both fiber systems terminated in their normal positions. We conclude that the sequence of fiber ingrowth does not determine the lamination of hippocampal afferents.

Afferent Pathways↗