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An analysis of the effects of retinoic acid and other retinoids on the development of adrenergic cells from the avian neural crest.

In the present work, we have investigated the role of all-trans-retinoic acid (all-trans RA), and several other natural and synthetic retinoids, in the development of adrenergic cells in quail neural crest cultures. Dose response studies using all-trans RA and 13-cis RA revealed a dose-dependent increase in the number of adrenergic cells in neural crest cultures. Similar dose response studies using RA isomers and other natural retinoids did not result in the same increases. In order to determine the receptor mediating the effects of all-trans RA in the neural crest, we tested several synthetic analogs which specifically bind to a particular RA receptor (RAR) subtype. We found that the compound AM 580, which activates the RAR-alpha, produced an increase in adrenergic cells similar to that seen with all-trans RA. The compound TTNPB, which activates all RAR subtypes, also resulted in an increase in adrenergic cells. We conclude that the increase in adrenergic cells seen with all-trans RA is mediated by RAR-alpha and possibly RAR-beta. To further define the actions of all-trans RA on the neural crest we incubated cultures with 5-bromo-2'-deoxyuridine (BrdU) to determine whether all-trans RA could affect the rate of proliferation. The results show that while all-trans RA did not increase the fraction of cells incorporating BrdU into their nuclei at early time points (24 h), it did increase BrdU incorporation by tyrosine hydroxylase (TH) positive cells at 5 days in culture. These findings demonstrate that the increase in adrenergic cells seen with all-trans RA in neural crest cultures is likely due to an increase in the proliferation of cells already expressing TH.