Immunocytochemical analysis of chromaffin cell proliferation in vitro.
The bromodeoxyuridine (BrdU) incorporation technique for immunocytochemical labeling of S-phase nuclei was optimized for the study of chromaffin cell proliferation. Sequential fixation in ethanol followed by paraformaldehyde, and the use of DNAse to render incorporated BrdU accessible to antibody, permitted permanent double staining for BrdU and tyrosine hydroxylase. The efficacy of the technique was demonstrated in microcultures of dissociated neonatal rat adrenal glands, in which chromaffin cells exhibited proliferative responses to nerve growth factor and fibroblast growth factor similar to those previously demonstrated by autoradiography. Growth factor responsiveness was observed in both serum-containing and serum-free medium.
Department of Pathology, Tufts University School of Medicine, Boston, Massachusetts 02111.
A S
AS
Tischler
L A
LA
Ruzicka
J C
JC
Riseberg
D000311:Adrenal Glands / Q000166:cytology* / Q000187:drug effects; D000818:Animals; D000911:Antibodies, Monoclonal; D001973:Bromodeoxyuridine; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002838:Chromaffin System / Q000166:cytology* / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D003851:Deoxyribonucleases; D005346:Fibroblast Growth Factors / Q000494:pharmacology*; D007150:Immunohistochemistry; D009414:Nerve Growth Factors / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D016196:S Phase; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis
D000911:Antibodies, Monoclonal; D003470:Culture Media; D009414:Nerve Growth Factors; D005346:Fibroblast Growth Factors; D014446:Tyrosine 3-Monooxygenase; D003851:Deoxyribonucleases; D001973:Bromodeoxyuridine