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Ontogeny of GABA-immunoreactive cells in the primate cerebellar cortex: comparison with somatostatin-immunoreactivity.

The distribution and ontogeny of GABA-immunoreactive cells were studied and compared with that of somatostatin-immunoreactivity in the primate cerebellar cortex. At embryonic day 80, we observed neither GABA-nor somatostatin-immunoreactive cells in the cerebellum. At embryonic day 110, a small number of GABA-immunoreactive cells was detectable in the granular layer only, and these cells seemed to be Golgi cells. At embryonic day 140, although almost all Purkinje cells were somatostatin-immunoreactive, a proportion of these cells was GABA-immunoreactive. At the newborn stage, most of the Purkinje cells were GABA-immunoreactive and almost all of them were also somatostatin-immunoreactive. During the postnatal stages, the number of somatostatin-immunoreactive cells decreased until postnatal day 60. At the adult stage, a large number of Purkinje cell bodies was faintly GABA-immunoreactive and a proportion of Purkinje cell dendrites was GABA-immunoreactive. In the aged animals (28 and 31 years old), a small number of Purkinje cell dendrites was GABA-immunoreactive. These findings suggest that a transition of phenotype from somatostatin to GABA occurred in Purkinje cells during development.

Age Factors↗

Quantitative ultrastructural localization of glutamate dehydrogenase in the rat cerebellar cortex.

Glutamate dehydrogenase is one of the main enzymes involved in the formation and metabolism of the neurotransmitter glutamate. In the present study we investigated the enzyme ultrastructurally in the cerebellar cortex, a region rich in well defined glutamatergic neurons, by pre-embedding immunocytochemical staining (peroxidase-antiperoxidase), as well as by post-embedding immunogold labelling employing a new system for quantitation and for specificity testing under the conditions of the immunocytochemical procedure. A new antiserum against immunologically purified bovine liver glutamate dehydrogenase or antibodies isolated from this by affinity chromatography were used in rats fixed by perfusion with aldehydes. The pre-embedding method displayed peroxidase reaction preferentially in mitochondria of astroglial cells (including the Bergmann glia). Mitochondria of neuronal tissue elements were usually free of peroxidase-reaction product. Extra-mitochondrial staining was not observed. The post-embedding immunogold method was employed to overcome penetration problems and allow semiquantitative analysis of localization and specificity. The highest densities of gold particles were found over the mitochondria in astroglial cell elements (including the Bergmann glia). Mitochondria in cell bodies of Bergmann glia had a lower particle density than those in astrocytic processes. In the latter, analysis of frequency distribution revealed no evidence of a population of mitochondria lacking glutamate dehydrogenase, but suggested the presence of populations with different levels of immunoreactivity. Comparison with the labelling of embedded bovine liver glutamate dehydrogenase indicated that the enzyme constitutes a high proportion (10%) of the total matrix protein of these mitochondria. A weaker but significant labelling was found in oligodendrocytes of the white matter. The labelling of mitochondria in neuronal elements including glutamatergic mossy fibre terminals was of the order of 15% of that in astroglial mitochondria. No difference was detected between glutamatergic neurons (mossy and parallel fibres, granular cells) and non-glutamatergic neurons (Purkinje cells). The particle density over non-mitochondrial areas was very close to background over empty resin. The results, obtained with different methods of tissue and antibody preparation, agree to show that the present form of glutamate dehydrogenase is restricted to mitochondria and preferentially localized in astrocytes.

Animals↗

Expression of proenkephalin mRNA in developing cerebellar cortex of the rat: expression levels coincide with maturational gradients in Purkinje cells.

The cellular localization of proenkephalin (PE) mRNA expression was systematically examined in midsagittal (vermal) sections of the developing rat cerebellar cortex by in situ hybridization. PE mRNA was initially detected in Golgi cells of postnatal day 7 (PND 7) rats and in each group thereafter. Moreover, PND 7 rats also displayed an intense layer of PE mRNA hybridization signal over the Purkinje cell layer. By PND 14, distinct cellular labeling was observed in a subpopulation of Purkinje cells in all lobules of the vermis except lobule III. At PND 7 and 14, the area and level of intensity of Purkinje cell associated PE mRNA hybridization signal followed a gradient that was most intense caudally but then decreased rostrally. At PND 21, the proportion of labeled Purkinje cells and the intensity of PE hybridization signal was evenly dispersed between the anterior and posterior lobules of the cerebellar vermis. PE hybridization signal was not detected in the developing neural cells of the external granular layer or the interneurons of the molecular layer in the vermis. These results indicate that the ontogeny of PE mRNA expression in Purkinje cells is developmentally regulated since levels of expression closely follow the chronological order of settling and maturation of these neurons. Based on prior evidence that endogenous opioids inhibit the growth of Purkinje cell dendrites and dendritic spines, PE expression is likely to be important for Purkinje cell maturation.

Animals↗

Electrical responses of the auditory area of the cerebellar cortex to acoustic stimulation.

Single unit activity from the VI and VII lobuli of the cerebellar vermis cortex was studied following acoustical stimulation with sound signals of different parameters. Cerebellar neurons, as compared to those from the auditory system, showed low selectivity to sound frequency, intensity and duration. However, about 2/3 of the neurons were selectively sensitive to interaural time and intensity differences; about 1/3 of neurons showed a specific response to signals simulating sound motion in a definite direction. Thus, cerebellar neurons seem to be mainly responsive to those sound parameters which are essential for sound localization.

Acoustic Stimulation↗

Propagation of parallel fiber volleys in the cerebellar cortex: a computer simulation.

We have studied the dynamic changes occurring during the propagation of active volleys through the parallel fiber (PF) tract in a portion of molecular layer of the cerebellar cortex, by means of a realistic computer simulation of the fibers (random distribution of PF origin; correlation between length and conduction velocity throughout depth). Surface electrical stimulations as well as mossy fiber stimulations of the PFs were mimicked. From a quantitative analysis of the propagation of the different types of volleys it appears that the constraints enforced by the morphological and functional characteristics of the PFs lead to an important spatial and temporal dispersion of the action potentials during their displacement along the folium. In particular, the time relationship between activity in the deep and superficial fibers as well as the number and location of active fibers at a given time (i.e. locus) will considerably vary during propagation. Moreover, in the case of a mossy fiber stimulation, there exists a striking asymmetry of the volleys travelling on each side of the site of stimulation. Therefore any information coded by the conjunctive activation of more than a few parallel fibers will be spatially and temporally dispersed in such a manner that successive postsynaptic Purkinje cells will receive very different combinations of inputs. This seems in favor of the hypothesis that these neurons function as coincidence detectors.

Action Potentials↗

Cytospectrophotometric study of the kinetics of histochemical reaction of GABA-transaminase in cryostat sections of rat cerebellar cortex.

A modification of the histochemical method for demonstration of GABA-transaminase is proposed. Substrate and cofactor concentrations are chosen on the basis of kinetic investigation in cryostat sections of the rat cerebellar cortex. Enzymatic reactions were measured by quantitative microspectrophotometry. Michaelis constants for alpha-oxoglutarate in the Purkinje cell layer and granular layer (Km 1.7 x 10(-3) M) and white matter (Km 3.8 x 10(-3) M) are found. It is shown that alpha-oxoglutarate in concentrations higher than 5.2 x 10(-3) M (1 mg/ml) suppresses the reaction in sections by competitive inhibition. The advisability of addition of malonate, PMS and cyanide to the incubation medium is confirmed. It is suggested that there are some isoenzymes of GABA-transaminase with predominant localization either in neurons or glia.

Animals↗