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The indirect antibody peroxidase-antiperoxidase technique was used to determine the laminar and lobular distribution of catecholaminergic afferents in the adult mouse, opossum, and cat cerebellum. A monoclonal antibody to tyrosine hydroxylase (TH) revealed a plexus of thin varicose fibers that exhibited a different density and distribution pattern for each species. In the cat, TH-immunoreactive fibers were sparsely distributed to all laminae, lobules, and nuclei of the cat cerebellum except for an area of elevated density in the ventral folia of lobules V and VI. In the opossum, TH-positive fibers were uniformly and densely distributed in the granule and Purkinje cell layers; they were more abundant in vermal lobules V-VI than in more anterior and posterior lobules, particularly I and X. Numerous TH-immunoreactive fibers were found in all four cerebellar nuclei of the opossum. In the mouse, TH-positive fibers formed a dense plexus within all cerebellar lobules, laminae, and nuclei. The mouse also exhibited numerous TH-immunoreactive Purkinje cells that were localized predominantly within vermal lobules VI-X, the paraflocculus, and flocculus. In addition to the interspecies differences in the distribution of catecholaminergic fibers within the cerebellum, comparison of this plexus to that previously described for serotonin in these species reveals that the relative densities and distribution patterns of catecholaminergic and serotoninergic fibers also vary between species. It is thus hypothesized that in each species a given monoamine has a unique net effect on cerebellar output that is determined by its effects on different neuronal populations within the cerebellum.

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Corticotropin releasing factor (CRF), localized in afferent inputs to the cerebellum, binds to two receptors defined as the Type 1 (CRF-R1) and the Type 2 (CRF-R2alpha). CRF-R1 has been localized to the cerebellum, as has a truncated isoform of CRF-R2alpha. Evidence for the presence of the full length isoform of CRF-R2alpha in the cerebellum is conflicting. We used RT-PCR, immunohistochemical, and physiologic techniques to resolve this conflict. RT-PCR data show low levels of CRF-R2alpha in the vermis and hemisphere of the cerebellum. These observations were confirmed by the Gene Expression Nervous System Atlas (GENSAT) database. A CRF-R2alpha antibody was used to determine the cellular distribution of the receptor in the cerebellum. The vast majority of the receptors are localized to Bergmann glial cells located throughout the cerebellum, as well as astrocytes in the granule cell layer. Neuronal labeling is present in sub-populations of Purkinje cells, Golgi cells, basket cells, and cerebellar nuclear neurons. Physiologic data show that urocortin II, which binds selectively to CRF-R2alpha, increases the firing rate of both Purkinje cells and nuclear neurons; this response can be blocked by the CRF-R2alpha-specific antagonist, antisauvagine-30. The present results confirm that CRF-R2alpha is present in the cerebellum and functions in circuits that modulate the firing rate of Purkinje cells and cerebellar nuclear neurons. A comparative analysis showed that the patterns of distribution of CRF-R1, CRF-R2alpha and CRF-R2alpha-tr are distinct. These data indicate that the CRF family of peptides modulates cerebellar output by binding to multiple CRF receptors.

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