IDENTIFICATION OF ACUTELY ISOLATED CELLS FROM DEVELOPING RAT CEREBELLUM

Immunocytochemical staining was used to identify nerve and glial cells from postnatal rat cerebelli in situ and following tissue dissociatio n. Purkinje cells were identified using antibodies for the calcium-bin ding proteins calbindin and PEP19. Purkinje cells isolated during the second postnatal week were 15-20 mum in diameter and relatively abunda nt and displayed thin perisomatic processes. These features were used to identify Purkinje cells with scanning electron microscopy, which re vealed extensive membrane infoldings. Golgi and nuclear cells were ide ntified using antibodies against rat-303 antigen. Pale, nuclear, and P urkinje cells were identified using antibodies for rat-302 antigen. Al though staining for rat-302 and rat-303 was weak during the second pos tnatal week, we were able to identify Golgi and pale cells even after tissue dissociation. Isolated Golgi cells were 8-10 mum in diameter an d fewer in number than Purkinje cells and did not counterstain with ca lbindin antibodies. Isolated pale cells were 8-10 mum in diameter, rar e, and resistant to calbindin antibodies. Isolated neurons from cerebe llar nuclei were not located with either 302 or 303 staining, suggesti ng that they remained in the tissue. Golgi-Bergmann cells and astrocyt es were identified using antibodies for glial fibrillary acidic protei n. Isolated glial cells were 12-15 mum in diameter, more numerous than Purkinje cells, and unstained with calbindin antibodies. With phase-c ontrast optics, glial cells appeared flatter than neuronal cell types and had acentric nuclei. These results demonstrate that specific cell types in developing rat cerebellum can be identified after acute isola tion, which should facilitate analysis of their endogenous properties. (C) 1994 Academic Press, Inc.