Stimulation of adrenergic development in neural crest cultures by a reconstituted basement membrane-like matrix is...

Previous work (Maxwell and Forbes: Development 101:767-776, 1987) has shown that an overlay of reconstituted basement membrane-like (RBM) gel dramatically increased the number of catecholamine-positive (CA+) cells which differentiated in neural crest cultures. We report here that this increase was inhibited when cultures were grown for 7 days in the presence of agents that elevate cAMP, such as 8-bromo-cAMP and 3-isobutyl-1-methylxanthine. The action of 8-bromo-cAMP was dose dependent with a half-maximal effect at about 50 microM. The development of CA+ cells was dramatically reduced when 8-bromo-cAMP was present from days 0-4 in vitro, but was relatively unaffected if 8-bromo-cAMP was present from days 4-7 in vitro. The development of tyrosine hydroxylase immunoreactive cells was also inhibited by 8-bromo-cAMP. The addition of 8-bromo-cAMP increased the number of melanocytes and resulted in either no change or only modest reductions in the number of E/C8 and neurofilament immunoreactive cells, indicating that the effect on CA+ cell ontogeny was selective. In contrast to the effect of 8-bromo-cAMP, addition of 8-bromo-cGMP did not inhibit CA+ cell development in the presence of the RBM gel nor did it stimulate CA+ cell development in the absence of the RBM gel overlay. Our results suggest that cAMP may be an important regulator of phenotypic expression in at least some neural crest cell lineages.(ABSTRACT TRUNCATED AT 250 WORDS)