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PubMed1997

Immunocytochemical localization of nicotinic acetylcholine receptor in the rat cerebellar cortex.

Although nicotinic acetylcholine receptors (nAChRs) have been reported to function in the cerebellar cortex, nicotinic acetylcholine receptor subtypes contributing to nicotinic currents in the cortex have not been well characterized at the subcellular level. This study deals with immunocytochemical localization of the nicotinic acetylcholine receptor alpha4 subunit using a monoclonal antibody against the alpha4 subunit. Alpha4-LI (alpha4-like immunoreactivity) was detected in the cell bodies of molecular, Purkinje cell and granular layers. In particular, the cell bodies of Purkinje cells were extensively immunostained. In Purkinje cells, alpha4-LI was found in perikarya mainly associated with rough endoplasmic reticulum, plasma membrane, and cytoplasmic matrix. At higher magnification, the immunoreaction product was densely localized along with somatic plasma membranes at the axo-somatic synapse and the plasma membranes at extrasynaptic regions of cell bodies. Alpha4-LI was also found in the axon terminals which form synapses with Purkinje cells. In the granular layer, somatic cell membranes of granular cells were immunostained. These morphological observations suggest that alpha4-containing nAChRs contribute nicotinic currents reported in Purkinje cells, and that presynaptic alpha4-containing nAChRs regulate the release of neurotransmitters on the axon terminals found near Purkinje cells.

Animals↗

PubMed1998

Apoptosis of undifferentiated progenitors and granule cell precursors in the postnatal human cerebellar cortex correlates with expression of BCL-2, ICE, and CPP32 proteins.

Naturally occurring apoptotic cells have been demonstrated in the postnatal cerebellum of rodents (Wood et al. [1993] Neuron 11:621-632; Krueger et al. [1995] J. Neurosci. 15:3366-3374). The nature of these cells differs among species: they are considered to be granule cells in mouse and astrocytes in rat. We labeled proliferating and apoptotic cells in the postnatal human cerebellar cortex by using antibodies against the Ki-67/proliferating cell nuclear antigen and the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method for fragmented DNA. We also immunocytochemically detected some proteins encoded by genes modulating apoptosis and specific markers of neuronal/glial differentiation. Proliferating cells were observed from birth to 4 months, representing 31-35% of cells within the external granular layer (EGL). Apoptotic cells were detected during the first 3 months and corresponded to 5-7% of EGL cells. Much lower percentages were calculated in other cortical layers and white matter. The balance between proliferation and apoptosis was quantitatively favorable to the latter during the first postnatal week. Expression of BCL-2, CPP32, and interleukin-1beta-converting enzyme (ICE) proteins was spatially and developmentally regulated in parallel with apoptosis. Apoptotic cells were often CPP32/ICE immunoreactive but negative for BCL-2. Some apoptotic cells were positive for vimentin and, less frequently, for alpha-internexin or type-III beta tubulin, but never expressed the glial fibrillary acidic protein. This study demonstrates that apoptosis is a significant phenomenon in early postnatal development of human cerebellar cortex and shares some of the regulatory mechanisms described in other vertebrates.

Antibodies↗

PubMed1995

Developmental changes of ganglioside expressions in postnatal rat cerebellar cortex.

We previously described the differential distribution of gangliosides in adult rat brain as detected by specific antibodies. We report here the distribution of gangliosides during the development of postnatal rat cerebellum by an immunofluorescence technique with mouse monoclonal antibodies (mAbs). Eleven mAbs that specifically recognize each ganglioside were used. Our study revealed that the expression of each ganglioside changed dramatically during the development. GD3 and O-Ac-GD3 were expressed intensely in the external granular layer at 1, 5, and 10 days, whereas GD2 was firstly detected in the internal granular layer at 5 days and GD1b WAS diffusely detected throughout all layers of the cerebellar cortex at early postnatal days. GD2 and GD1b were more intensely expressed in the granular layer at 20, 30, and 80 days, suggesting that premature granule cells expressed GD3 and its derivative, O-Ac-GD3, whereas mature granule cells express GD2 and GD1B intensely. On the other hand, GM1 was exclusively detected in the external granular layer and the molecular layer at 1 and 5 days. The staining sites spread gradually from these outer layers into the internal granular layer and the white matter after 10 days. The positive cells in the external granular layer and the molecular layer appeared to be Bergmann glial cells and their radially ascending cytoplasmic processes. The intensity of the staining in these specialized astroglial cells decreased gradually during postnatal days. In contrast, the expression of GQ1b was very faint at birth, but gradually increased during the development and was detected intensely in the internal granular layer, particularly in the cerebellar glomeruli in adulthood, suggesting that GQ1b expression may be associated with synapse-related structures. The developmental changes of the expression of other gangliosides were also recognized in the postnatal rat cerebellum. These results suggest that specific gangliosides may play an important role in regulating the early events responsible for the orderly formation of the cerebellar cortex.

Animals↗

PubMed1978

A comparison of the inhibitory effects of taurine and GABA on identified Purkinje cells and other neurons in the cerebellar cortex of the rat.

The microiontophoretic application of taurine and GABA was studied in the cerebellar cortex of the rat. Both taurine and GABA produced a dose-dependent depression of spike frequency of cerebellar neurons. GABA (2-42 nA, mean 27 nA) induced an inhibition of spike discharge on all 138 cells tested, including 29 Purkinje cells. Taurine (60-200 nA, mean 108 nA) induced an inhibition of spike discharge on 93 of the 106 cerebellar neurons tested, including inhibition on 22 of 25 Purkinje cells. Iontophoretic application of bicuculline and picrotoxin antagonized the inhibitory effects of both GABA and taurine on Purkinje cells as well as on cerebellar neurons in general. Strychnine did not antagonize the inhibition of either GABA or taurine. Simultaneous application of taurine and GABA produced a synergistic inhibitory effect on the firing rate of Purkinje cells. Taurine, in contrast to GABA, appeared to be more depressant when applied in the Purkinje cell dendritic zone than when applied near the soma. The data are discussed in terms of taurine functioning as a neurotransmitter in the cerebellum of the rat and having receptor sites distinct from those for GABA.

Aminobutyrates↗

PubMed2005

Thyroid hormone regulates Galphai1 gene expression in the rat cerebellar cortex during post-natal development.

Thyroid hormone regulates the expression of G protein in tissues such as fat and heart. In the brain, very little information is available relative to the regulation by thyroid hormone of G proteins. Here, we show that the expression of the Galphai1 gene is induced by thyroid hormones in the rat cerebellum during development. Hence, the levels of Galphai1 transcripts and protein were decreased in the cerebellum of hypothyroid neonates. In situ hybridization studies showed that the neurons of the cerebellar cortex, particularly Purkinje cells, were affected. Surprisingly, and in contrast with the in vivo stimulatory effect described above, thyroid hormone repressed the activity of the rat Galphai1 promoter in vitro, suggesting that the effect of this hormone in the cerebellum is indirect. In this regard, we present data suggesting that the transcription factor C/EBPbeta could be implicated. First, there are active CEBP binding sites in the Galphai1 promoter. Second, we have found a diminished DNA binding activity of hypothyroid nuclear proteins to a Galphai1 promoter sequence containing a C/EBP binding site. Third, this complex is likely to contain C/EBPbeta protein as it is displaced by specific anti-C/EBPbeta antibodies. Finally, there is a significant decrease in the C/EBPbeta protein content in the hypothyroid cerebellar cortex.

Animals↗

PubMed1989

The GABAergic neurones of the cerebellar nuclei in the rat: projections to the cerebellar cortex.

The presence of gamma-aminobutyric acid (GABA) in the neurones of the cerebellar nucleocortical pathway is here reported. The pathway was identified by retrograde tracer and the GABA content was revealed immunohistochemically. It was found that most of the neurones giving rise to the reciprocal, non-reciprocal and symmetrical projections are indeed GABA-immunoreactive. They were observed in all the subdivisions of the nucleus medialis, of the nucleus interpositus and of the nucleus lateralis sending axons respectively to the sagittal zones A, C1-3 and D of the cerebellar cortex. The nucleus vestibularis lateralis and the related sagittal zone B were devoid of such projections.

Animals↗

PubMed1983

Responses of fastigial single neural units to transfolial electrical stimulation of the cerebellar cortex in the cat.

In anesthetized cats, we recorded the responses of single neural units in the fastigial nucleus to electrical stimulation of the posterior vermal surface of the cerebellar cortex. Stimulus frequencies of 0.3 Hz, 10 Hz, and 200 Hz were used and the responses were similar at each frequency. Two principal responses were found: suppression-activation sequences and suppression followed by return to baseline. The former were found in the caudal portions of the nucleus and the latter along with the majority of nonresponsive units were found in the rostral portions. An area of overlap was found in the central portion of the nucleus.

Animals↗

PubMed1977

The pathogenesis of acute selective necrosis of the granular layer of the human cerebellar cortex.

PH determinations by means of a sturdy, spear-electrode inserted in the cerebellar tissue immediately after necropsy were carried out in 60 patients suffering from different disorders and dying in hospital, and from 25 individuals killed instantly by violent accidents. A correlation was found between the presence of NGL and low pH levels in the cerebellum (average pH 5.56); subjects without NGL changes showed an average pH OF 6.08. No correlation was found between the pH levels and the time after death, or between pH and any specific disease. The so-called acute selective necrosis of the granular layer of the human cerebellar cortex (NGL), is considered from the present studies to be a phenomenon of postmortem origin. Its relationship to certain specific diseases can be explained by the fact that in these conditions there is a higher postmortem acidity, known to promote autolysis.

Aged↗

PubMed2004

Compartmentation of the mouse cerebellar cortex by sphingosine kinase.

Classic cerebellar anatomy is based on the characteristic array of lobes and lobules. However, there is substantial evidence to suggest that more fundamental architecture is built around arrays of parasagittal stripes, which encompass both the inputs and outputs of the Purkinje cells (PCs). Sphingosine kinase (SPHK) is an enzyme that converts sphingosine (Sph) into sphingosine-1-phosphate (S1P). Recent reports have indicated that ceramide, Sph, and S1P play a role in cell survival, growth, and differentiation in several cell types, including neurons. In this study, we examined the localization of SPHK in the mouse cerebellum by using immunohistochemistry. Anti-SPHK immunoreactivity appeared in the cerebellar molecular layer and the PC membranes. The staining pattern is striped. In the molecular layer, the staining pattern probably reflects dendritic spines and dendrites. By electron microscopy, peroxidase reaction product was deposited within dendrites especially along the plasma membranes near spines. Seen at higher magnification, the staining was in and near the postsynaptic complexes. By double immunostaining, the striped pattern of SPHK expression was shown to be identical to that revealed by anti-zebrin II, although the subcellular distribution within PC's is not. This is the first demonstration of the cerebellar compartmentation of an enzyme related to lipid metabolism, and as such, it provides an insight into the roles of SPHK and formation of S1P. The selective expression of SPHK in the zebrin II-immunoreactive PCs may explain their resistance to cell death when ceramide metabolism is disrupted, as in the acid sphingomyelinase knockout model of Niemann-Pick type A/B disease.

Amino Acid Sequence↗

PubMedDate not supplied

[Possibility of derivation of convulsive potentials in the cerebellar cortex].

Explore the source record for details and available documents.

Cerebellar Cortex↗

PubMed1977

Ultrastructural pattern of acetylcholinesterase distribution in the cerebellar cortex of the quail.

The ultrastructural localization of acetylcholinesterase (AChE) was studied in the cerebellar cortex of the quail by means of histochemical method. The greater amount of AChE was detected at leve of the molecular layer in the intracellular spaces between parallel fibers and between parallel fibers and dendritic terminals. Many neurons showed intracellular localization of enzyme activity: the AChE positive neurons were all Golgi cells, most stellate the basket cells and different aliquots of Purkinje and granule cells. The enzymatic activity was usually localized in the cisternae of endoplasmic reticulum, in the nuclear envelope (but this last localization was not present in Purkinje cells- and sometimes in the Golgi apparatus; reaction granules were usually scarce in the different dendritic branches ramifying in the molecular layer. On the basis of the ultrastructural pattern of AChE distribution, some considerations are developed on the methodological aspects concerning the reliability of histochemical methods, the differences recorded at light and electron microscope level, the problems related to extracellular localization of enzyme, the difficulty of establishing a precise correlation between AChE localization in a cerebellar neuron and its possible cholinergic and/or cholinoceptive nature.

Acetylcholinesterase↗

PubMed1986

Responses of cerebellar cortex to electrical stimulation of the glossopharyngeal nerve in the frog.

In the frog cerebellar cortex, electrical stimulation of the glossopharyngeal (IXth) nerve induced negative field potentials with a peak latency of 57 ms whose distribution was bilateral with ipsilateral predominance. The site where the maximum negativity was induced by IXth nerve stimulation was histologically located within the molecular layer near the Purkinje cell layer. In extracellular recording, electrical stimulation of the IXth nerve induced complex and/or simple spike discharges of Purkinje cells. Such evoked potentials and unitary spikes in the cerebellum were attributed to the excitation of the IXth nerve afferents of higher threshold which are mainly composed of fibers sensitive to taste stimulation. These results suggest that gustatory information projects to the cerebellum, as well as those of other kinds of senses, such as touch, visual and auditory sensation.

Animals↗

PubMed1988

Quantitative study of granule and Purkinje cells in the cerebellar cortex of the rat.

The numerical densities of granule and Purkinje cells in the cerebellar cortex of the rat were determined by stereological methods. The density of Purkinje cells in our fixed material was 1,018 +/- 39 per mm2 (mean +/- s.e.m.) of Purkinje cell layer and that of granule cells 1.92 +/- 0.03 x 10(6) per microliter of granular layer. The total area of Purkinje cell layer was 332 mm2 and the volume of granular layer was 48 microliters. The rat cerebellum therefore contains 3.38 x 10(5) Purkinje cells and 9.2 x 10(7) granule cells, from which there are 274 granule cells for each Purkinje cell. The density of granule cells and the density of parallel fibers in the molecular layer observed in a companion study indicate that the average length of a parallel fiber is around 5 mm.

Animals↗

PubMed1993

Synaptic and non-synaptic immunolocalization of GABA and glutamate acid decarboxylase (GAD) in cerebellar cortex of rat.

The existence of a large number of GABA receptors in the cerebellar molecular layer, and the observation of numerous punctate immunoreactive deposits of GABA synthesizing enzyme (GAD) throughout this layer, could indicate the existence of numerous axon terminals that may be involved in neurotransmission modulated by GABA. These axon terminals may be different from those considered classically as cerebellar GABAergic axon terminals. Therefore, we have reinvestigated the localization of GABA- and GAD-immunoreactivities in the cerebellar cortex of the rat with the PAP method, using different antisera obtained from rabbits immunized with GABA, baclofen and GAD. The results observed in our investigation have demonstrated GABA- and GAD-immunoreactivities in the axon terminals considered classically as GABAergic, as well as in others which, until now, have not been considered GABAergic. This fact leads us to think that the distribution of GABA or molecules structurally similar to GABA is far more extended than previously thought in the cerebellum. We have also observed both GABA- and GAD-immunoreactivities within dendrites and glial cells. These facts suggest us a possible extrasynaptic release of GABA.

Animals↗

PubMed1992

Ultrastructural localization of inositol 1,4,5-trisphosphate 3-kinase in rat cerebellar cortex.

Subcellular localization of inositol 1,4,5-trisphosphate 3-kinase in the rat cerebellar cortex was studied immunohistochemically using a monoclonal antibody. Electron microscopy revealed intense immunoreactivity in the dendritic spines of Purkinje cells forming synapses with the parallel fibers, climbing fibers and recurrent collaterals of Purkinje cell axons. The labelling was associated with the hypolemmal cisternae, surrounding matrix and plasmalemma including the postsynaptic densities. Weaker immunoreactivity was present in the dendritic spines of basket cells and in certain segments of Purkinje cell recurrent collaterals. The postsynaptic regions of the dendritic trunks of Purkinje and basket cells were negative. These results indicate that inositol 1,4,5-trisphosphate 3-kinase is distributed amongst the spines of various synaptic relations with different electrophysiological properties, and that axon terminals of certain cell types are another functional site for the enzyme.

Animals↗

PubMedDate not supplied

[The structural-functional organization of the endoplasmic reticulum in the neurons of the rat cerebellar cortex].

Electron microscopic investigation of neuronal elements in the cerebellar cortex of rats revealed the existence of septal reticulum in axonal collaterals of Purkinje cells, in axons of basket cells and in mossy fibers. It was shown that this reticulum acts as a calcium depot. The presence in neuroplasm of the developed system of evenly distributed reticulum, which forms relatively isolated compartments and acts as the calcium depot may account for local regulation of rearrangements in the cytoskeleton. Wide presentation of the septal reticulum in the nervous cells of both invertebrates and vertebrates indicates functional determination of this structure.

Animals↗

PubMed2001

Purkinje cells in the adult cat cerebellar cortex possess a perineuronal net of proteoglycans.

The Purkinje cells in the adult cat cerebellar cortex were found to possess perineuronal proteoglycans which could be stained with our fine cationic iron colloid and Fujita's highly concentrated aldehyde fuchsin, and digested by chondroitinase ABC/keratanase/ heparitinase and hyaluronidase. The Purkinje cells are surrounded by some collagenous elements which are stained with Gömöri's ammoniacal silver and digested by collagenase. The Purkinje cells also express nerve cell surface glycoproteins which are labeled with lectin Vicia villosa agglutinin and digested by a double treatment with collagenase and endo-alpha-N-acetylgalactosaminidase. Sole digestion by endo-alpha-N-acetylgalactosaminidase never erased the lectin labeling of the nerve cell surface glycoproteins. These findings suggest that the collagenous elements mediate the linkage of the perineuronal proteoglycans to the nerve cell surface glycoproteins. It is presumed that in mice and rats, the perineuronal nets of proteoglycans and nerve cell surface glycoproteins of the Purkinje cells are so thin or coarse that they can not be sufficiently visualized under the light microscope.

Animals↗

PubMed1990

Deposition and role of thrombospondin in the histogenesis of the cerebellar cortex.

The patterns of deposition of thrombospondin (TSP), a trimeric extracellular matrix glycoprotein, were determined during the initial establishment of the external granule cell layer and the subsequent inward migration of granule cells forming the molecular and (internal) granule cell layers. The early homogeneous deposition of TSP became restricted to the rhombic lip in the region of granule cell exit from the neuroepithelium, and was present between migrating granule cells. During the later inward migration of granule cells, little TSP was associated with dividing granule cells; it was enriched in premigratory granule cells. With the cessation of migration, TSP was lost except in association with fasciculating axons in the molecular layer where staining persisted briefly. At the EM level, TSP was associated with the leading process of granule cells as they associated with Bergmann glial cells and migrated through the molecular layer. TSP was present within granule cell axons; Purkinje cells and their dendrites, as well as Bergmann glial fibers and endfeet were negative for TSP. When anti-TSP antibodies were added to explant cultures of cerebellar cortex during active granule cell migration, a dose-dependent inhibition of migration was observed. In control cultures, granule cells migrated into the (internal) granule cell layer, while granule cells exposed to anti-TSP antibodies were arrested within the external granule cell layer. These results suggest that TSP plays an important role in the histogenesis of the cerebellar cortex by influencing granule cell migration.

Aging↗