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 dissociation. Purkinje cells were identified using antibodies for the calcium-binding proteins calbindin and PEP19. Purkinje cells isolated during the second postnatal week were 15-20 microns in diameter and relatively abundant and displayed thin perisomatic processes. These features were used to identify Purkinje cells with scanning electron microscopy, which revealed extensive membrane infoldings. Golgi and nuclear cells were identified using antibodies against rat-303 antigen. Pale, nuclear, and Purkinje cells were identified using antibodies for rat-302 antigen. Although staining for rat-302 and rat-303 was weak during the second postnatal week, we were able to identify Golgi and pale cells even after tissue dissociation. Isolated Golgi cells were 8-10 microns in diameter and fewer in number than Purkinje cells and did not counterstain with calbindin antibodies. Isolated pale cells were 8-10 microns in diameter, rare, and resistant to calbindin antibodies. Isolated neurons from cerebellar nuclei were not located with either 302 or 303 staining, suggesting that they remained in the tissue. Golgi-Bergmann cells and astrocytes were identified using antibodies for glial fibrillary acidic protein. Isolated glial cells were 12-15 microns in diameter, more numerous than Purkinje cells, and unstained with calbindin antibodies. With phase-contrast 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 isolation, which should facilitate analysis of their endogenous properties.
Hockberger
P E
PE
Department of Physiology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Yousif
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L
Nam
S C
SC
D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000502:physiology*; D002469:Cell Separation; D048429:Cell Size / Q000502:physiology; D002531:Cerebellum / Q000166:cytology*; D005260:Female; D008297:Male; D008855:Microscopy, Electron, Scanning; D008858:Microscopy, Phase-Contrast; D009457:Neuroglia / Q000166:cytology* / Q000648:ultrastructure; D011247:Pregnancy; D011689:Purkinje Cells / Q000166:cytology* / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley