Selective retrograde labelling of the rat olivocerebellar climbing fiber system with D-[3H]aspartate

Selective retrograde labelling of the olivocerebellar climbing fiber system with D-[3H]aspartate has been observed in the rat, and the results have implications for the identification of a transmitter candidate as well as the neuroanatomical understanding of these cerebellar afferents. Microinjections of D-[3H]aspartate (50 nl, ca 10-2 M) were made into various parts of cerebellar cortex. Survival times were 6, 12 or 24 h. Pronounced diffusion of the tracer resulted in large injection sites. Within the zone of injection, glial elements were labelled over background. Most granule cells exposed to the tracer were unlabelled; the small numbers demonstrating labelling were believed to have been injured by the micropipette penetration. Beneath injection sites, large numbers of well-labelled nerve fibers appeared in the white matter and could be followed through the brainstem to the contralateral inferior olive, where labelled perikarya were found. After the inferior olivary neurons had been effectively destroyed with 3-acetylpyridine, evidence of cerebellar afferent labelling with D-[3H]aspartate was missing. Retrograde labelling of the olivocerebellar system was also observed after superfusion of the vermis with D-[3H]aspartate at concentrations in the range of Km for high affinity uptake (10(-5) or 10(-4) M, for 2 h). Mossy fiber or monoaminergic afferents to the cerebellum were never labelled with D-[3H]aspartate. The distribution of labelled cells in the olivary subnuclei after injections in different cerebellar areas was in line with the olivocerebellar organization previously described in the cat. Moreover, it was demonstrated that fibers from the different subnuclei follow different routes through the brainstem towards the cerebellum. Labelling of climbing fiber collaterals in uninjected parts of cerebellum indicated that some of the retrogradely migrating D-[3H]aspartate was directed in anterograde direction at axonal branching points. Collaterals were demonstrated in all deep cerebellar and Deiters' nuclei, and the results of intranuclear injections suggested that virtually every olivary neuron sends collaterals to these nuclei. Intracortical collaterals were organized in sagittal zones. Midline injections into the anterior lobe and VI lobule labelled collaterals in several zones of the posterior lobe spinal area and uninjected parts of the anterior lobe vermis. Hemispheral injection into copula pyramidis labelled collaterals in two prominent bundles in the anterior lobe.(ABSTRACT TRUNCATED AT 400 WORDS)