Cholinergic innervation of the cerebellum of rat, rabbit, cat, and monkey as revealed by choline acetyltransferase activity and immunohistochemistry

The cholinergic innervation of the cerebellar cortex of the rat, rabbit, cat and monkey was studied by immunohistochemical localization of choline acetyltransferase (ChAT) and radiochemical measurement of regional differences in ChAT activity. Four antibodies to ChAT were used to find optimal immunohistochemical localization of this enzyme. These antibodies selectively labeled large mossy fiber rosettes as well as finely beaded terminals with different morphological characterization, laminar distribution within the cerebellar cortex, and regional differences within the cerebellum. Large “grape‐like” classic ChAT‐positive mossy fiber rosettes that were distributed primarily in the granule cell layer were concentrated, but not exclusively located in three separate regions of the cerebellum in each of the four species studied: (1) The uvula‐nodulus (lobules 9 and 10); (2) the flocculus‐ventral paraflocculus, and (3) the anterior lobe vermis (lobules 1 and 2). No intrinsic cerebellar neurons were labeled. No cells in either the inferior olive (the origin of cerebellar climbing fibers) or in the locus coeruleus (an origin of noradrenergic fibers) were ChAT‐positive. Thin, finely beaded axons, similar to cholinergic axons of the cerebral cortex of the rat, were observed in both the granule cell layer and molecular layer of the cerebellar cortex of the rat, rabbit and cat. The regional differences in ChAT‐positive afferent terminations in the cerebellar cortex was for the most part confirmed by regional measurements of ChAT activity in the rat, rabbit, and cat. The three cholinergic afferent projection sites correspond to regions of the cerebellar cortex that receive vestibular primary and secondary afferents. These data imply that a subset of vestibular projections to the cerebellar cortex are cholinergic.