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PubMed1983

Mitochondria in the postnatally developing rat cerebellar cortex: a morphological and biochemical study.

Ultrastructural changes during development in the proliferating cells of the external granular layer and the granule cells of the internal granular layer of the rat cerebellar cortex were examined. Changes in the number of free ribosomes, development of membrane systems, nuclear appearance and the number of cristae and matrix density of mitochondria suggested that both the round and elongated cells of the external granular layer and the granule cells of the internal granular layer were at same stage of maturation at any given age. Quantitatively, the relative volumes of cytoplasm and of mitochondria in the granule cells of internal granular layer were significantly higher than in the proliferating external granular layer cells; however, mitochondrial density and its increase during development were not significantly different in the two cell populations. The activity of the mitochondrial enzyme, succinate dehydrogenase, was also similar in ultrastructurally preserved and metabolically competent perikaryal fractions enriched in replicating external granular layer cells, granule cells and Purkinje cells. These findings emphasize the similarities between proliferating external granular layer cells and granule cells of internal granular layer. They lend support to the view that these cell types at the ages examined, represent a continuum of maturation towards full neuronal differentiation.

Animals↗
PubMed1985

[Cerebellar cortex: simulation of recurrent collateral inhibition with the aid of a set of neuronal automata. I. Model and global results].

In view of a study on the transfer function of the cerebellar cortex a modelization of each of the various subsystems was undertaken. The recurrent collateral inhibition existing between Purkinje cells was mimicked by means of an assembly of neuronal automata (NA) temporally evolving at random through three states ("silent", "tonic" and "phasic" and interacting with simple rules. In such an assembly--by comparison with a control group of independent NA--a drastic modulation of activity appears. First the total of the mean number of state shifts is increased by more than 10%. This increase is clearly hierarchical with a maximal effect for the "silent" state. Second the average duration of each state also varies, although in a contrasted manner. Third, and moreover, a clear spatial cooperativity emerges. Indeed all the individually coupled NA are in the same "silent" state at identical moments for more than 5.5% of the total running time. A cyclic repetition of this spatiotemporal cooperativity is apparent. The emergence of these collective properties--which can not be deduced linearly from the unitary elements--introduces a parameter of order leading to a coherent functioning of the system.

Animals↗
PubMed1985

Basket cell development in the normal and hypothyroid rat cerebellar cortex studied with a monoclonal anti-neurofilament antibody.

We have used a monoclonal antibody against an antigenic determinant of the 210-kdalton neurofilament protein to study basket cell maturation in rat cerebellar cortex. Neurofilament immunoreactivity first appears in basket cells at postnatal day 12 and mature axonal "pinceaux" are present at postnatal day 17. There are large differences in the rate of maturation from lobe to lobe which do not fully correspond to the rate of Purkinje cell differentiation. In hypothyroid rats the expression of the neurofilament antigen by basket cells is almost completely suppressed.

Animals↗
PubMed1995

Developmental expression and functional characterization of the 4C5 antigen in the postnatal cerebellar cortex.

The monoclonal antibody 4C5 recognizes a neuron-specific surface antigen (4C5 antigen) in the CNS and PNS of the rat. In the present study we investigated the expression of 4C5 antigen in the developing cerebellum of the rat and the functional role of this molecule during cerebellar morphogenesis. Immunoblotting and immunohistochemistry in sections of cerebellar cortex revealed an age-dependent decrease in the expression of the 4C5 antigen. In cerebellar primary cell cultures, 4C5 immunoreactivity was detected both on granule and on Purkinje neurons. Granule cell migration was inhibited in cerebellar explants derived from 8-day-old rats and cultured for 2 days in the presence of antibodies against the 4C5 antigen. Electron microscope immunocytochemistry revealed that in 8-day-old rat cerebellum, 4C5 immunoreactivity was localized on the cell bodies of granule neurons in the external and internal granular layers and on parallel fibers in the developing molecular layer as well as at contact sites between these cellular elements. It was not detected on Bergmann glia. These results suggest strongly that the 4C5 antigen is involved in granule cell migration during cerebellar development, possibly via neuron-neuron interactions.

Aging↗
PubMed1985

Monoclonal antibodies reveal sagittal banding in the rodent cerebellar cortex.

We have produced two monoclonal antibodies against polypeptide-associated antigens of developing rat cerebellum. One antibody recognizes an antigen associated with synaptic vesicles and another binds to a polypeptide which is restricted to the cytoplasm of a subset of cerebellar Purkinje cells. Both antibodies reveal the biochemical differentiation of the rodent cerebellar cortex into antigenically distinct sagittal zones.

Animals↗
PubMed1995

NMDA receptor expression in the mouse cerebellar cortex.

A detailed, light microscopic study on the distribution of the N-methyl- D-aspartate receptor subunit 1 (NMDAR1) was carried out with immunohistochemistry and in situ hybridization on the cerebellar cortex of the mouse. With a monoclonal antibody, labeling of Purkinje cell bodies varied from intense to negative, while heavy dendritic staining was limited to the proximal dendrites (unlike the rat, which also had heavily stained distal dendrites). In the granular layer, the cell bodies and and the dendritic shafts of Golgi II cells were only moderately stained, but very intense labeling was associated with granule cell bodies, and with their dendrites and dendritic endings in the glomeruli. The mossy and climbing fibers were negative. In situ hybridization with a cRNA probe showed levels and spatial distributions of NMDAR1 mRNA consistent with the immunolabeling pattern, in that signals were strongest in the granular and Purkinje cell layers and relatively low or absent in the molecular layer and white matter. The findings are consistent with the hypothesis that NMDAR1 may be especially well concentrated at the synaptic target sites of the mossy and climbing fibers. In the mouse, NMDAR1 at the parallel fiber sites associated with Purkinje cell spiny branchlets may differ from the rat in its level of expression or in its molecular configuration.

Animals↗
PubMed2006

[Local blood flow in the dorsal hippocampus and cerebellar cortex in progeny of iodine-deficient rats].

Experimental studies show depressive behavior in rats caused by hypothyroidism and antidepressant effect of thyroid hormones. The present study analyses changes in the hippocampal and cerebellar cortex local blood flow in the progeny of rats suffering from iodine deficiency before mating and during the whole period of gestation. The diet with very low iodine content results in a decrease of local blood flow in both brain structures, but the greatest changes were observed in hippocampus. Addition of the iodine to the diet eliminates the above blood flow changes.

Animals↗
PubMed1991

Cytology and organization of reactive astroglia in human cerebellar cortex with severe loss of granule cells: a study on the ataxic form of Creutzfeldt-Jakob disease.

In order to investigate the cellular basis of human astrogliosis, we have selected the cerebellar cortex because it provides a relatively simple and geometrical organization of both neuronal and glial populations. A pathological system with severe and progressive loss of granule cells was studied: the ataxic form of Creutzfeldt-Jakob disease, where the tissue geometry is minimally disturbed. The quantitative study revealed a drastic reduction in the numerical density of granule cells in the Creutzfeldt-Jakob disease cerebellum, and a significant increase in the numerical density of astrocytes. Karyometric analysis showed that the nuclear area was significantly greater in reactive astroglial cells than in normal astroglia. Glial fibrillary acidic protein immunocytochemistry revealed astroglial hypertrophy, but the geometry and spatial domains of astroglial subtypes were strictly preserved. Vimentin expression was detected in Bergmann glia and in certain astrocytes of the granular layer. Ultrastructural analysis showed that reactive astroglia had large nuclei, with expanded interchromatinic regions which contained clusters of interchromatin granules and nuclear bodies, and prominent reticulate nucleoli. In the cytoplasm, hypertrophied bundles of intermediate filaments were observed, some of them associated with the nuclear envelope. Numerous adhering and gap junctions were also found among reactive astroglial cells. Perivascular glial processes showed a terminal web of intermediate filaments and a conspicuous plasmalemmal undercoat. Interendothelial tight junctions were preserved. Our results suggest that the severe loss of granule cells induces a highly ordered astroglial response which tends to preserve the geometry of the astroglial scaffold, the domains of each astroglial subtype, the neuronal microenvironmental conditions and the efficiency of the blood brain barrier, in order to promote neuron survival.

Astrocytes↗
PubMed1996

Electron microscopic demonstration of intracelluar promethazine accumulation sites by a precipitation technique: application to the cerebellar cortex of the mouse.

A method is described that allows electron microscopic identification of the phenothiazine neuroleptic promethazine after supravital intracardiac injection of high drug concentrations (greater than or equal to 3 %). The cerebellar cortex of the mouse was used for the investigation. This procedure is based on simultaneous fixation of drug and tissue by immersion in a paraformaldehyde-glutaraldehyde solution with the addition of phosphomolybdic acid. The electron microscopic investigation revealed that the drug could easily be identified as an electron-dense precipitate. Subpopulations of neurons exhibited a higher affinity for the drug than others, but no preference for any nerve cell type was detected. Closer study showed that promethazine accumulated primarily within the cytoplasm, where it was bound to the endoplasmic reticulum. Furthermore, promethazine storage was observed within mitochondria and vesicular structures. Drug binding was also seen in the regions of synapses but without any predilection for these structures. Because this precipitation technique also appears to be well-suited for the ultrastructural identification of other drugs, it offers multiple possibilities for application.

Animals↗
PubMed1976

[Repercussions of hypothyroidism on the synaptogenesis of the rat cerebellar cortex: quantitative study of the thyroid hormone effect according to the age (author's transl)].

The corrective action of Thyroxine on the deficit of the synaptic density of the molecular layer of the cerebellar cortex was studed quantitatively, in the rat made hypothyroid by propylthiouracile (P.T.U.). In the normal as in the hypothyroid rat, the increase of the synaptic density does not extend over the fifth postnatal week. In the PTU treated animals, during the 2 first postnatal weeks, the administration of 0,25 mug/d during the third and 0.50 mug/d during the fourth postnatal week brings the synaptic density back to normal. In the PTU treated animals, during the 5 postnatal weeks, the administration during a week only of 0.50 mug/d of LT4 between 1 and 14 days or, of 1 mug/d between 15 and 28 days, increases, but does not return to normal the synaptic density. On the contrary, the administration of 1 mug/d of LT4 during 1 or even 2 consecutive weeks 28 days later, remains without effect on the synaptic density. After the end of the fourth postnatal week, the deficit of the synaptic density becomes irreversible.

Age Factors↗
PubMed1993

Reactive astroglia-neuron relationships in the human cerebellar cortex: a quantitative, morphological and immunocytochemical study in Creutzfeldt-Jakob disease.

In order to investigate the role of neuron-glia interactions in the response of astroglial to a non-invasive cerebellar cortex injury, we have used two cases of the ataxic form of Creutzfeldt-Jakob disease (CJD) with distinct neuronal loss and diffuse astrogliosis. The quantitative study showed no changes in cell density of either Purkinje or Bergmann glial cells in CJ-1, whereas in the more affected CJ-2 a loss of Purkinje cells and an increase of Bergmann glial cells was found. The granular layer in both CJD cases showed a similar loss of granule cells (about 60%) in parallel with the significant increase in GFAP+ reactive astrocytes. GFAP immunostaining revealed greater reactivity of Bergmann glia in CJ-2 than in CJ-1, as indicated by the thicker glial processes and the higher optical density. Granular layer reactive astrocytes were regularly spaced. In both CJD cases there was strict preservation of the spatial arrangement of all astroglial subtypes--Fañanas cells, Bergmann glia and granular layer astrocytes. Reactive Fañanas and Bergmann glial cells and microglia/macrophages expressed vimentin, while only a few vimentin+ reactive astrocytes were detected in the granular layer. Karyometric analysis showed that the increase in nuclear volume in reactive astroglia was directly related with the level of glial hypertrophy. The number of nucleoli per nuclear section was constant in astroglial cells of human controls and CJD, suggesting an absence of polyploidy in reactive astroglia. Ultrastructural analysis revealed junctional complexes formed by the association of macula adherens and gap junctions. In the molecular layer numerous vacant dendritic spines were ensheathed by lamellar processes of reactive Bergmann glia. Our results suggest that quantitative (neuron/astroglia ratio) and qualitative changes in the interaction of neurons with their region-specific astroglial partners play a central role in the astroglial response pattern to the pathogenic agent of CJD.

Astrocytes↗
PubMed1990

Immunocytological localization of the highly polysialylated form of the neural cell adhesion molecule during development of the murine cerebellar cortex.

The expression of the highly polysialylated form of the neural cell adhesion molecule (N-CAM)--the so-called embryonic N-CAM (E-N-CAM)--was investigated in the developing and adult mouse cerebellar cortex by immunohistology and immunocytology at the light and electron microscopic levels. E-N-CAM was never (from embryonic day 14 to postnatal day 15) detectable in the germinal zone of neuroblasts destined to form or forming the external granular layer and was only observed once small cerebellar interneurons had become postmitotic before the beginning of granule cell migration. Granule cells expressed E-N-CAM on cell bodies, axons, and leading and trailing processes also during migration but ceased to reveal detectable levels of E-N-CAM at the end of migration after having reached their final position in the internal granular layer. Other cerebellar cell types, such as Purkinje cells, Bergmann glia, astrocytes, oligodendrocytes, and most prominently, stellate and basket cells, also expressed E-N-CAM, but became E-N-CAM-negative during the third and fourth postnatal weeks, coinciding with overt cessation of cerebellar histogenesis. Thus, except for neuroblasts, E-N-CAM appeared characteristic of growing and moving cellular structures, in agreement with the notion that the highly polysialylated form of N-CAM is less adhesive than the adult form.

Animals↗
PubMed1996

Purkinje cell lineage and the topographic organization of the cerebellar cortex: a view from X inactivation mosaics.

We utilized a strain of mice, derived from a radiation mutagenesis experiment and carrying an activity-attenuated allele of the X-linked enzyme glucose-6-phosphate dehydrogenase (G6PD), to analyze the development of the cell lineage leading to cerebellar Purkinje neurons. Due to random X inactivation during early embryonic development, X- linked genes can be used to distinguish between clonally related populations of cells in X inactivation mosaics. Following histochemical staining for G6PD activity, the numeric proportions of Purkinje cells expressing either the wild-type or the mutant enzyme and the spatial distribution of these cellular phenotypes and their relation to anatomically and genetically defined cerebellar compartments were analyzed. Our data suggest that cerebellar Purkinje neurons originate from a limited pool of some 129 precursors. The size of this pool is different from the one derived from chimeric mice, allowing us to deduce the relative timing of Purkinje cell lineage restriction. Our data also show that Purkinje neurons of distinct lineage are extensively intermingled within the cerebellar cortex. Together, these findings suggest both a role for cell-cell communication in the development of genetically defined cerebellar compartments and a temporal window during which such cellular interactions may take place.

Animals↗
PubMed1984

Immunocytochemical demonstration of alpha-tubulin modification during axonal maturation in the cerebellar cortex.

Previous light microscopic immunocytochemical studies using two monoclonal antibodies that recognise alpha-tubulin (YOL/34 and YL1/2) but differ in their isotypic specificity have shown that the unmyelinated parallel fiber axons in the cerebellar cortex are labeled with only one of the antibodies (YOL/34). We now show that at 10 d postnatally the parallel fibers are labeled with both antibodies, and that during development YL1/2 (but not YOL/34) immunoreactivity disappears progressively from parallel fibers in the lower regions of the molecular layer upwards towards the external germinal layer. By approximately 28 d postnatally, the differential staining pattern of parallel fibers by the antibodies is established throughout the molecular layer. The time course, light microscopic, and ultrastructural staining distribution corresponds to a progressive change in alpha-tubulin immunoreactivity as the parallel fibers form synaptic contacts. This modification of alpha-tubulin (which was not observed in Purkinje cell dendrites or Bergmann glia) may be related to the formation of a basic isotype of alpha-tubulin within parallel fiber axons at maturation.

Animals↗
PubMed1983

Cerebellar cortex ultrastructure in ataxia-telangiectasia.

An ultrastructural study of a biopsy fragment from the cerebellar cortex of a patient with ataxia-telangiectasia was performed. There was a sharp reduction in the number of Purkinje cells and an increase in the number of Golgi epithelial cells. Despite the degeneration of Purkinje cell dendrites, the volumetric density of parallel fiber varicosities and the number of synapses in the molecular layer were only slightly reduced. No qualitative nor quantitative abnormalities were noted in the granular layer. An analysis is made of the severe changes in cerebellar circuitry of this patient and their probable functional implications.

Ataxia Telangiectasia↗
PubMed1987

The polypeptide PEP-19 is a marker for Purkinje neurons in cerebellar cortex and cartwheel neurons in the dorsal cochlear nucleus.

This light and electron microscopic immunocytochemical study shows that the polypeptide PEP-19, a presumptive calcium binding protein specific to the nervous system, represents an excellent marker for cerebellar Purkinje cells and dorsal cochlear nucleus (DCoN) cartwheel cells. The polypeptide clearly reveals the entire populations of both types of neurons, including their complete dendritic and axonal arborizations. Other PEP-19 containing neurons in the two regions display weak immunoreactivity restricted to the cell body or to cell body and principal dendrites. Electron microscopic localization of PEP-19-like immunoreactivity reveals similarities between this polypeptide, parvalbumin, and a 28K vitamin D-dependent calcium binding protein. However, calmodulin, which is expressed in both Purkinje and granule cells, may differ from PEP-19. Similarities between the organization of the cerebellar cortex and the DCoN superficial layers have been known for some time, with several types of neurons in one system having their presumed homologue in the other. These data provide further support for the proposed structural and functional homology between Purkinje and cartwheel neurons, and establishes PEP-19 as a useful marker for examining degeneration of these two neuronal populations in murine cerebellar mutants.

Animals↗
PubMed2002

Compartmentation of the rabbit cerebellar cortex.

The cytoarchitecture of the adult rabbit cerebellum is revealed by using zebrin II/aldolase c immunocytochemistry in both wholemount and sectioned material. Zebrin II is expressed by approximately half of the Purkinje cells of the cerebellar cortex. In most regions these form a symmetrical array of zebrin II positive and negative parasagittal bands. Four transverse expression domains are identified in the vermis: (1) an anterior zone, comprising four narrow bands, one at the midline and three laterally to either side, extending throughout the anterior lobe to the primary fissure; (2) a central zone with broad immunoreactive bands separated by narrow zebrin II negative bands that disappear caudally to leave no apparent compartmentation; (3) a posterior zone with prominent alternating zebrin II positive and negative bands; and (4) a nodular zone in which all Purkinje cells express zebrin II. In the hemispheres a striped topography is found in lobules HVI, HVII, and crus I, and all Purkinje cells are zebrin II+ in the flocculus and paraflocculus. Because of its importance for the classical conditioning of the eyeblink response, we made a detailed analysis of lobule HVI of the hemisphere. The immunocytochemical data show a complex substructure within HVI with three prominent zebrin II positive bands (probably homologous with P4a+, P4b+, and P5+ of rodents) separated by two zebrin II negative regions (P4- and P4b-). Thus, the organization of the rabbit cerebellum is consistent with the patterns described previously for rat, mouse, and opossum and suggests that there may be a common ground plan for the mammalian cerebellum.

Animals↗