Transcriptional silencing of glucocorticoid-inducible phenylethanolamine N-methyltransferase expression by sequential signaling events.

Specific arrays and timing of environmental cues including glucocorticoids, neurotrophic factors and intracellular messengers influence phenotype expression in developing chromaffin cells or sympathetic neurons. Although the two lineages are closely related, only adrenergic chromaffin cells express phenylethanolamine N-methyltransferase (PNMT), the enzyme that synthesizes epinephrine, while neurons and noradrenergic chromaffin cells are PNMT-negative. It remains unclear to what extent the ability to express PNMT is determined by environmental cues versus intrinsic heterogeneity already present in ganglionic and adrenal precursors. Mouse pheochromocytoma (MPC) cell lines are a model for studying adrenergic differentiation. In two MPC lines that exhibit up to 1000-fold induction of PNMT mRNA by dexamethasone, pretreatment with glial cell line-derived neurotrophic factor (GDNF) and/or the cyclic AMP analog cpt-cAMP markedly blunts or abrogates PNMT inducibility. PNMT suppression occurs without apparent neuronal differentiation in one of the MPC lines and in normal adult mouse chromaffin cell cultures. Our results establish transcriptional suppression by cAMP as a mechanism for regulating PNMT expression in both normal and neoplastic mouse chromaffin cells. However, contrast between large increases in PNMT mRNA levels and low stimulation of promoter activity suggests that modulation of mRNA degradation also plays an important role. Clarification of mechanisms that regulate these two processes in MPC cells may provide insight into developmental mechanisms governing expression and maintenance of the adrenergic phenotype.