Brain calcium/calmodulin-dependent protein kinase type II, a multimeric 600-650 kDa enzyme composed of alpha- (50 kDa) and beta/beta' (60 and 58 kDa) subunits, may be formed by alpha- and beta-subunits combining in variable proportions in different types of neurons. This study presents evidence, using cerebella from mutant mice, that the alpha-subunit displays a restricted localization in the rodent cerebellum, being detectable only in Purkinje cells. Immunocytochemical analysis of normal rat cerebellum with an antibody selective for the alpha-subunit confirmed that this subunit was detectable only in Purkinje cells. In contrast, the beta/beta'-subunits appeared to be present in all types of cerebellar mutants examined. These results indicate that different cells of the cerebellum express distinct isozymic forms of the multifunctional calcium/calmodulin-dependent protein kinase type II. It appears that Purkinje cells primarily contain an isoenzyme formed by both alpha- and beta/beta'-subunits, and that non-Purkinje cells contain an isoenzyme formed primarily by beta/beta'-subunits.