Nerve growth factor and dexamethasone specify the catecholaminergic phenotype of cultured rat chromaffin cells: dependence on developmental stage.

Antibodies to epinephrine (E) and bovine phenylethanolamine N-methyltransferase (PNMT) have been used to monitor the regulation of the E- and PNMT-immunoreactive chromaffin cell phenotypes by dexamethasone (DEX) and nerve growth factor (NGF). The cells were isolated from 1-, 10- and 30-day-old (D1, D10 and D30) rat adrenal glands and grown for 4 days on a polyornithine substratum. DEX (10(-5)M) supported the survival of 90% of the cells from all postnatal ages studied. In contrast, only 45% (D1), 33% (D10) and 60% (D30) of the chromaffin cells had survived after 4 days in control cultures or when treated with NGF (100 ng/ml). Throughout this study numbers of E-immunoreactive cells were approximately 10% larger than those of cells stained by anti-PNMT antibodies irrespective of the treatments applied. 55% of the cells isolated at D1 and 79% of the cells at D10 were stained by anti-E antibodies. The proportion of E-positive cells was constant in D1 cultures carried for 4 days, while E-immunoreactive cells dropped to 63% in cultures from D10. At D1 and D10 DEX and NGF had opposite effects on the portions of E-positive cells, DEX increasing and NGF decreasing their relative numbers, 66% of the chromaffin cells isolated at D30 displayed E-specific immunoreactivity. DEX caused a significant increase (to 74%), while both NGF-treated and control cultures exhibited a decrease in the relative numbers of E-immunoreactive cells.(ABSTRACT TRUNCATED AT 250 WORDS)