Time course of enkephalin mRNA and peptides in cultured rat adrenal medulla

Explantation of rat adrenal medullae to organ culture results in dramatic changes in enkephalins and catecholamines that are similar to the changes seen in vivo in response to denervation, which eliminates transsynaptic impulse activity. We have used rapid and sensitive solution hybridization methods to measure preproenkephalin (PPenk) mRNA and total cellular RNA in samples from rat tissues and adrenal medullary explants. The profiles of adrenal medullary PPenk mRNA, enkephalin-containing (EC) peptides, total cellular RNA and catecholamines [epinephrine (epi) and norepinephrine (norepi)] were measured during 14 days of organ culture. After 8 h in culture, total RNA had declined by 60%, epi and norepi declined 80 to 85% and EC peptides by 50% while the amount of PPenk mRNA per gland increased by 400%. Between 8 h and 14 days total RNA and catecholamine levels remained constant while PPenk mRNA increased to a peak of 85 +/- 10 (S.E.M.) pg/gland at 2-4 days, a value that was 80 times greater than the zero time (preculture) values. EC peptide levels lagged behind the increase in PPenk mRNA and reached a peak of 25 +/- 4 (S.E.M.) pmol Met-enkephalin equivalents/gland at 4 days that was 80 times greater than zero time values. Both PPenk mRNA and EC peptides declined in parallel between 4 and 14 days. The ratio of the copies of proenkephalin (Penk) peptide to PPenk mRNA was estimated to be 25,000 at the time of explantation and after 4 days in culture. From steady-state kinetics half-life estimates of 9.6 h for PPenk mRNA and 14.7 h for Penk peptide were obtained.(ABSTRACT TRUNCATED AT 250 WORDS)