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M Frotscher

Publications and source records attributed to M Frotscher.

At least 127 records · Page 7Linked to original sources
PubMed1993

GABAergic innervation of the rat fascia dentata: a novel type of interneuron in the granule cell layer with extensive axonal arborization in the molecular layer.

By using the combined Golgi/electron microscopy (EM) technique and postembedding immunocytochemistry for gamma-aminobutyric acid (GABA), we describe a novel type of local circuit neuron in the rat fascia dentata that gives rise to an axon profusely ramifying in the dentate molecular layer. The relatively small ovoid cell body (long axis 12-15 microns) is located directly underneath the granular layer. From both poles of the cell body dendritic processes emerge that enter the molecular layer and hilar region, respectively. The apical dendrites traverse the granular layer, invade the molecular layer, and branch in the same way as granule cell dendrites. Some branches reach the hippocampal fissure. Thus, the apical dendrites of these neurons may receive a similar input pattern as the granule cells. The dendrites are smooth, occasionally bearing varicosities. A few spines are regularly observed. The axon originates from the apical dendrite and traverses the molecular layer horizontally for up to 500 microns. It gives off numerous collaterals that are distributed throughout the entire width of the molecular layer and only rarely enter the granule cell layer. Electron microscopy of the cell body of gold-toned neurons revealed the well-known fine-structural characteristics of nonpyramidal neurons, i.e., an indented nucleus with nuclear inclusions and large aggregations of endoplasmic reticulum. Apical as well as basal dendrites are densely covered with presynaptic boutons, mainly forming asymmetric synapses. The axon terminals of these cells form symmetric synapses with dendritic shafts and, to a lesser extent, with spines. These symmetric synapses, together with the results of our GABA postembedding immunocytochemical study, suggest that this cell is a GABAergic inhibitory neuron that almost exclusively innervates the dentate molecular layer. Together with data from the literature on dentate axoaxonic cells (which innervate the axon initial segments of the granule cells) and GABAergic basket cells (which innervate the granule cell somata and proximal dendrites in the granular layer), the present results indicate that there is a lamination of the GABAergic innervation of the fascia dentata corresponding to the well-known segregated termination of entorhinal and commissural afferents to this region.

Animals↗
PubMed1993

Are the fine-structural characteristics of mouse hippocampal mossy fiber synapses determined by the density of mossy fiber axons?

Heritable variation of mossy fiber synapses in hippocampal region CA3 was studied in the two inbred mouse strains C3H and CPB-K. Previous Timm studies had shown a larger mossy fiber projection in C3H mice. In contrast, synaptic boutons of CPB-K mice were larger in size and perimeter and were contacted by more dendritic spines than in C3H mice. These results point to an inverse relationship between the size of the mossy fiber projection and the number of spine synapses formed by an individual mossy fiber bouton. Thus, the fiber density of a projection may be crucial for the actual morphology of the synaptic contacts formed.

Animals↗
PubMed1993

Spiny nonpyramidal neurons in the CA3 region of the rat hippocampus are glutamate-like immunoreactive and receive convergent mossy fiber input.

There is increasing evidence that the various types of hippocampal nonpyramidal neurons control the principal cells in different ways. In the present study a type of spiny nonpyramidal cell in stratum lucidum of rat hippocampal region CA3 was studied by Golgi impregnation. Three Golgi-impregnated and gold-toned neurons of this type were further analyzed by electron microscopy and postembedding immunocytochemistry. The dendrites of these bipolar neurons seemed to be restricted to stratum lucidum and ran parallel with the mossy fibers that terminate in this layer. A characteristic feature of this neuron is the presence of long, thin spines on both cell body and dendrites. Although these dendrites were exposed to a large number of mossy fibers, no thorny excrescences were formed which are characteristic postsynaptic elements of CA3 pyramidal neurons for synaptic contact with the mossy fibers. Semithin sections of Golgi-impregnated and gold-toned stratum lucidum cells displayed immunoreactivity of the cell body region for glutamate but not for GABA. A fine-structural analysis of gold-toned sections revealed a large cell body with numerous cytoplasmic organelles and an indented nucleus. Numerous asymmetric synapses were found on dendritic shafts as well as on the long, thin somatic and dendritic spines. Usually, several presynaptic boutons contacted a single spine. The majority of these asymmetric spine synapses were probably of mossy fiber origin, although no giant mossy fiber synapses were formed. The long spines were contacted by much smaller en passant synapses of preterminal axons. In contrast, giant mossy fiber boutons were found presynaptic to dendritic shafts and cell bodies of these cells. Our morphological analysis of a glutamate-immunoreactive, GABA-negative type of nonpyramidal neuron that receives convergent mossy fiber input suggests that the impulse flow within the "trisynaptic pathway" is more complex than previously assumed.

Afferent Pathways↗
PubMed1993

Developmental neurotrophin expression in slice cultures of rat hippocampus.

Using reverse transcription in combination with the polymerase chain reaction, the developmental expression of neurotrophins in organotypic slice cultures of rat hippocampus was investigated. Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) mRNA levels after different time periods in vitro were compared with equivalent developmental stages in vivo. Our results show that neurotrophin expression occurs in hippocampal slice cultures with a similar time course as observed in the developing hippocampus in vivo. Thus, the development of neurotrophin expression in the hippocampus does not seem to be dependent on specific extrinsic afferents.

Afferent Pathways↗
PubMed1993

Development of the rat septohippocampal projection: tracing with DiI and electron microscopy of identified growth cones.

The factors determining the development of specific fiber tracts in the central nervous system as well as the interactions of growth cones with the surrounding micromilieu are largely unknown. Here we investigated the ontogenetic development of the septohippocampal projection in the rat with the lipophilic carbocyanine dye DiI which is transported anterogradely and retrogradely in neurons and can be applied to fixed embryonic tissue. Photoconversion of anterogradely labeled fibers allowed us to study individual growth cones by electron microscopy. The first axons originating from the septal complex were found in the hippocampus as early as on embryonic day (ED) 19, reaching the fimbrial pole of the hippocampus on ED 18. However, on ED 17 we consistently found retrogradely labeled cells in the hippocampus, indicating that the development of the hippocamposeptal projection precedes that of the septohippocampal projection. On ED 19, the majority of the axons directed toward the hippocampal formation passed the hippocampus and grew further into the subicular complex and entorhinal cortex. These axons gave off collaterals that invaded the hippocampus proper. A fairly adult pattern of the septohippocampal projection was reached on postnatal day 10, although may growth cones were still found. A comparative analysis of individual growth cones found in the fimbria and the hippocampus proper revealed no striking differences in their morphology. Electron microscopic analysis showed that growth cones in the fimbria were mainly contacted by other axons, whereas growth cones in the hippocampus had contact with all available elements. This may indicate that growing septohippocampal fibers are guided by axons of the earlier formed hippocamposeptal projection. In the hippocampus proper, other cues, probably derived from the target itself, may guide the septohippocampal axons to their appropriate target cells.

Afferent Pathways↗
PubMed1993

Loss of layer-specific astrocytic glutamine synthetase immunoreactivity in slice cultures of hippocampus.

Glutamine synthetase (GS) supposedly inactivates the excitatory neurotransmitter glutamate. By using immunocytochemistry for GS, we recently demonstrated a layer-specific, perisynaptic distribution of GS-immunoreactive astrocytes and their processes in perfusion-fixed rat hippocampi. Highest levels of immunoreactivity were found in well defined termination zones of glutamatergic hippocampal afferents. In the present study we analysed the developmental aspect of this neuron-glia interaction by using hippocampal slice cultures lacking all extrinsic afferents. Under these conditions, no layer-specific distribution of astrocytic GS immunoreactivity could be demonstrated. This suggests that the laminated distribution of GS immunoreactivity is formed in parallel with the segregated termination of hippocampal afferents. Thus, there is no predetermined pattern of GS-containing astrocytes playing a role in the segregation of extrinsic fibres. The ultrastructural localization of GS immunoreactivity in fine astrocytic processes around asymmetric, probably glutamatergic excitatory spine synapses confirms earlier in situ findings, which suggests that this arrangement is a global phenomenon of glutamatergic systems.

Animals↗
PubMed1993

Transneuronal changes in dendrites of GABAergic parvalbumin-containing neurons of the rat fascia dentata following entorhinal lesion.

The perforant path fibers from the entorhinal cortex form synapses with both granule cells and GABAergic, parvalbumin-containing (PARV) nongranule cells. The authors recently reported a persistent reduction of PARV-positive dendrites in the termination zones of entorhinal fibers in the hippocampus proper and fascia dentata after lesion of the entorhinal cortex. In the present study the authors analyzed the effects of de-entorhination on the ultrastructure of postsynaptic PARV-positive dendrites in the molecular layer of the fascia dentata. PARV immunocytochemistry was performed 2, 8, 55, and 360 days after an ipsilateral entorhinal lesion and, for comparison, 10 days after an ipsilateral fimbria-fornix transection that disconnects the hippocampus from its septal and commissural afferents. Two days after entorhinal lesion, the authors observed swelling of the tissue close to the hippocampal fissure. Adjacent distal dendritic tips of PARV-positive dentate neurons appeared bloated and reduced in number. Reduction of PARV-positive dendrites in the former perforant path termination zone persisted 55 days after entorhinal lesion and could still observed after postlesional survival times for 1 year. Degenerating axon terminals were still present 55 days following lesion and PARV-positive dendrites exhibited abnormal invaginations. Fimbria transection did not result in similar dendritic changes in PARV-positive neurons. The results indicate a long-lasting process of reorganization in the molecular layer of the fascia dentata following entorhinal lesion and persisting changes in the morphology of PARV-immunoreactive dendrites. Entorhinal fibers seem to play a specific role for the maintenance of these dendrites, since similar changes did not occur following removal of septal and commissural fibers.

Animals↗
PubMed1993

Formation of the septohippocampal projection in vitro: an electron microscopic immunocytochemical study of cholinergic synapses.

Cholinergic neurons in the medial septum/diagonal band complex project to the hippocampus and fascia dentata and establish characteristic types of synapses on a variety of target neurons. At present we do not know the principles that underlie the development of this projection and the formation of the cholinergic synapses. Here we have used co-cultured slices of septum and hippocampus of one- to six-day-old rat pups to study the development of the septohippocampal pathway and the formation of cholinergic synapses on hippocampal target neurons in vitro. Slices of septum and hippocampus were incubated together for 10-46 days applying the roller-tube technique. The fluorescent dye dioctadecyltetramethylindocarbocyanine perchlorate and histochemical staining for acetylcholinesterase labeled many fibers connecting both explants. Combined light- and electron-microscopic immunocytochemistry for choline acetyltransferase, the acetylcholine-synthesizing enzyme, revealed multipolar immunopositive neurons with long aspiny dendrites in the septal culture. Numerous varicose immunoreactive, supposedly cholinergic fibers could be followed from the septal to the hippocampal culture where they ramified and formed a three-dimensional network. As in situ, cholinergic terminals formed characteristic symmetric synapses on cell bodies, spines and, most often, on dendritic shafts of the hippocampal target neurons. No immunoreactive fibers and synapses were observed in single cultures of hippocampus. These results demonstrate that the cholinergic septohippocampal projection develops in vitro and that similar types of cholinergic synapses are established on co-cultured hippocampal target neurons as observed in situ.

Acetylcholinesterase↗
PubMed1992

Application of the Golgi/electron microscopy technique for cell identification in immunocytochemical, retrograde labeling, and developmental studies of hippocampal neurons.

In this study the Golgi/electron microscopy (EM) technique has been used for an analysis of the fine structure, specific synaptic connections, and differentiation of neurons in the hippocampus and fascia dentata of rodents. In a first series of experiments the specific synaptic contacts formed between cholinergic terminals and identified hippocampal neurons were studied. By means of a variant of the section Golgi impregnation procedure, Vibratome sections immunostained for choline acetyltransferase, the acetylcholine-synthesizing enzyme, were Golgi-impregnated in order to identify the target neurons of cholinergic terminals in the hippocampus. It could be shown with this combined approach that cholinergic septohippocampal fibers form a variety of synapses with different target structures of the Golgi-impregnated and gold-toned hippocampal neurons. In this report cholinergic synapses on the heads of small spines, the necks of large complex spines, dendritic shafts, and cell bodies of identified dentate granule cells are described. The variety of cholinergic synapses suggests that cholinergic transmission in the fascia dentata is a complex event. Next, the Golgi/EM technique was applied to Vibratome sections that contained retrogradely labeled neurons in the hilar region of the fascia dentata following horseradish peroxidase (HRP) injection into the contralateral hippocampus. With this combined approach some of the hilar cells projecting to the contralateral side were identified as mossy cells by the presence of retrogradely transported HRP in thin sections through these Golgi-impregnated and gold-toned neurons. Our findings suggest that the mossy cells are part of the commissural/associational system terminating in the inner molecular layer of the fascia dentata. They are mainly driven by hilar collaterals of granule cell axons that form giant synapses on their dendrites. Finally, the Golgi/EM procedure was used to study the differentiation and developmental plasticity of hippocampal and dentate neurons in transplants and slice cultures of hippocampus. Under both experimental conditions, the differentiating neurons are deprived of their normal laminated afferent innervation but develop their major cell-specific characteristics including a large number of postsynaptic structures (spines). As revealed in thin sections of gold-toned identified cells, all these spines formed synapses with presynaptic boutons suggesting sprouting of the transplanted and cultured neurons, respectively. Altogether, the present report demonstrates the usefulness of the Golgi/EM technique, particularly of the section impregnation procedure, for a variety of studies requiring the identification of individual neurons at the ultrastructural level.

Animals↗
PubMed1992

Fine structure of rat septohippocampal neurons: I. Identification of septohippocampal projection neurons by retrograde tracing combined with electron microscopic immunocytochemistry and intracellular staining.

In this report the normal dendritic organization and fine structure of identified septohippocampal projection neurons is described as a prerequisite for a time course analysis of retrograde changes in these neurons following axotomy (see Naumann et al., J. Comp. Neurol. 325:219-242, 1992). Septohippocampal projection neurons were retrogradely labeled by injection of the fluorescent tracer Fluoro-Gold into the hippocampus. Next, retrogradely labeled cells in Vibratome sections of the medial septum/diagonal band complex were intracellularly stained with the fluorescent dye Lucifer Yellow (LY). Photooxidation of LY resulted in a stable electron-dense reaction product, which allowed us to study these double-labeled neurons by electron microscopy. Another series of sections containing retrogradely labeled neurons were immunostained for choline acetyltransferase (ChAT) or parvalbumin (PARV). In this way the fine structure of two different chemically characterized subpopulations of septohippocampal neurons could be compared with that of the LY-injected neurons. Intracellular filling of retrogradely labeled neurons with LY stained the cell body and the entire dendritic arbor. Essentially, three classes of neurons could be distinguished, i.e., bipolar cells, multipolar neurons, and an intermediate group. All these neurons displayed smooth, often varicose dendrites lacking spines. Mainly located close to the midline, there was a group of cells with only very few if any LY-stained dendrites. In the electron microscope, the double-labeled neurons were easily identified by numerous electron-dense lysosomes associated with transported Fluoro-Gold and the diffuse reaction product resulting from photooxidation. They displayed fine-structural characteristics as previously described for cholinergic neurons. In fact, our fine-structural analysis of ChAT-positive Fluoro-Gold-labeled neurons, but also of back-filled PARV-positive cells, gave very similar results. All these neurons had infolded nuclei, abundant cytoplasmic organelles, and a few axosomatic synapses. Thus, a plain electron microscopic study does not allow one to distinguish between subpopulations of septohippocampal projection neurons.

Animals↗
PubMed1992

Fine structure of rat septohippocampal neurons: II. A time course analysis following axotomy.

Previous light microscopic immunocytochemical studies with antibodies against transmitter-synthesizing enzymes have suggested that septohippocampal neurons undergo retrograde degeneration following transection of their axons by cutting the fimbria-fornix. However, a fine-structural analysis of the degeneration process in these cells is lacking so far. Here we have identified septohippocampal neurons by retrograde tracing with Fluoro-Gold. Thereafter, the fimbria-fornix was transected bilaterally. Fine-structural changes in prelabeled septohippocampal neurons were then studied after varying survival times up to 10 weeks. Examination under the fluorescence microscope of Vibratome sections through the septal region revealed numerous retrogradely labeled cells after all survival times following axotomy. These neurons were then intracellularly injected with the fluorescent dye Lucifer Yellow in order to stain their dendritic arbor. Many cells were found after each survival time that displayed characteristics of septohippocampal neurons in control rats (see Naumann et al., J Comp Neurol 325:207-218, 1992). In addition, increasing with survival time, there were many shrunken neurons with a reduced dendritic arbor. Representative examples of both normal appearing and shrunken neurons were photoconverted for subsequent electron microscopic analysis. Relatively few signs of neuronal degeneration were found at each survival time analyzed. The majority of cells, including the heavily shrunken ones, displayed fine-structural characteristics of normal neurons. However, a few degenerating neurons and reactive glial cells were present in all survival stages. We conclude that axotomized septohippocampal projection neurons cease the expression of transmitter-synthesizing enzymes and shrink, but many more cells survive for extended periods of time without target-derived neurotrophic factor than was assumed in previous light microscopic studies.

Animals↗
PubMed1992

Reduction of posttraumatic transneuronal "early gene" activation and dendritic atrophy by the N-methyl-D-aspartate receptor antagonist MK-801.

The removal of a major hippocampal afferent system, the glutamatergic fibers from the entorhinal cortex, results in transneuronal changes in postsynaptic inhibitory neurons using gamma-aminobutyric acid (GABA) as a neurotransmitter. This study shows that these transneuronal alterations are reduced by the selective N-methyl-D-aspartate (NMDA) receptor antagonist (+)-MK-801. Thus, systemic injection of (+)-MK-801 prior to and after entorhinal lesion abolishes the retraction of distal dendrites from the termination zones of degenerating entorhinal fibers and reduces the swelling of their distal segments. Also, entorhinal lesion results in the appearance of c-fos protein-like immunoreactivity in hippocampal neurons and glial cells, which again is blocked by (+)-MK-801 administration. These data suggest that NMDA receptor-mediated neurotoxicity due to postlesional glutamate elevation results in early gene expression and in transneuronal dendritic changes. Similar processes may play a role in Alzheimer's disease, since there is a severe degeneration of the glutamatergic entorhino-hippocampal projection in this neurodegenerative disorder.

Afferent Pathways↗
PubMed1992

Parvalbumin-containing nonpyramidal neurons in intracortical transplants of rat hippocampal and neocortical tissue: a light and electron microscopic immunocytochemical study.

Previous immunocytochemical studies have shown that GABAergic nonpyramidal neurons of the rat hippocampus survive in intracerebral transplants. However, information is still lacking about the dendritic organization and the input synapses of these cells as well as their capacity to express the calcium-binding protein parvalbumin (PARV) under transplant conditions. In the present study, a monoclonal antibody against PARV was used to examine the dendritic morphology and the synaptic organization of parvalbumin-containing GABAergic neurons in hippocampal and dentate transplants. In addition, parvalbumin-containing nonpyramidal neurons were studied in neocortical transplants to compare the differentiation of grafted allocortical and neocortical nonpyramidal neurons. Tissue blocks of hippocampus and fascia dentata and of the parietal neocortex were taken from late embryonic rats (E 21 and E 16, respectively) and were transplanted into a cavity in the somatosensory cortex of young adult rats. After 3.5 or 7 months survival, the recipient brains were fixed by perfusion and immunostained for PARV. As in the hippocampal formation in situ, PARV-containing neurons in the hippocampal transplants were observed within and in the vicinity of the pyramidal and granule cell layer. In neocortical transplants, PARV-immunoreactive cells were distributed in all parts of the transplant with dendrites extending in various directions. In both hippocampal and neocortical transplants, immunoreactive dendrites were smooth and displayed the characteristic regular varicosities known from in situ studies of these cells. Numerous unlabeled terminals as well as a few immunoreactive boutons established synapses on the immunoreactive dendrites. PARV-positive terminals formed the typical pericellular baskets around the immunonegative cell bodies of pyramidal neurons and granule cells in the transplants. They established symmetric synapses with cell bodies and proximal dendrites. Synapses on axon initial segments were absent or rare. Our results demonstrate that allocortical as well as neocortical nonpyramidal neurons transplanted to the neocortex of adult recipients survive transplantation, express the calcium-binding protein parvalbumin, and develop a cell-specific morphology.

Animals↗
PubMed1992

Identified septohippocampal neurons survive axotomy: a fine-structural analysis in the rat.

Previous studies have indicated that interruption of the connections between the medial septum and hippocampus by cutting the axons results in degeneration and death of the projecting septal neurons. However, in these studies cell death has been inferred primarily from the loss of immunoreactivity for transmitter-specific enzymes. In the present study, we labeled septohippocampal projection neurons by retrograde tracing and then cut their axons. Subsequent intracellular injection of prelabeled cells revealed the morphology of the soma and dendrites and allowed us to examine the ultrastructure of these neurons. A large number of septohippocampal neurons survived even 10 weeks after axotomy, suggesting that axotomized septohippocampal neurons survive for considerable periods beyond the time at which they stop expressing transmitter-specific immunoreactivity. Survival of axotomized neurons is a prerequisite for pharmacological interference aimed at reactivating transmitter expression, axonal re-growth, and the eventual reintegration into functionally relevant circuitries.

Animals↗
PubMed1992

Reorganization of input synapses of parvalbumin-containing neurons in the rat fascia dentata following entorhinal lesion.

Removal of ipsilateral entorhinal afferents to the fascia dentata results in a retraction of postsynaptic parvalbumin (PARV)-containing dendrites of GABAergic neurons from the outer molecular layer. This study analyzes the reorganization of input synapses of these identified neurons following deafferentation. The density of synaptic input (total length of synaptic membrane specializations) of PARV-immunostained dendrites increased by 34% in the outer molecular layer of the fascia dentata 8 days, and by 21% 55 days following lesion when compared with unoperated controls. Eight days postlesion this increase was mainly due to an enlargement of synaptic membrane specializations of single terminals whereas 55 days after the lesion there was an increase in the number of synapses on the identified dendrites. Our results suggest expansion of terminals of remaining afferent systems (i.e. commissural fibers) in the early postlesional period and reactive synaptogenesis (i.e. de novo formation of synaptic contacts) on PARV-positive dendrites after long survival time. This increased innervation may be of functional importance as it might compensate for the reduction of the receptive field of the PARV-positive, supposedly inhibitory neurons in the postlesional dentate gyrus.

Animals↗