Role of retinoic acid in mouse neural crest cell development in vitro

This study examines the role of retinoic acid (RA) in mouse neural crest cell development in culture. We have compared the effects of RA on the developmental behavior of mouse neural crest cells from different axial levels, that is the mesencephalic (cranial) and sympathoadrenal (trunk) levels by colony assay using antibodies against cell-type-specific markers. In control cultures in the absence of RA, the efficiency of colony formation by cranial neural crest cells was considerably lower than in colony cultures of trunk neural crest cells. Pulse-labelling experiments using 5-bromo-2'-deoxyuridine (BrdU) showed that the proportion of neural crest cells that incorporated BrdU was significantly smaller in day 5 cultures of cranial neural crest cells in the absence of RA compared to cultures from the trunk level. However, BrdU-incorporation matched the labelling observed in trunk crest cell cultures when RA was added to the culture medium. The efficiency of colony formation and the proportion of BrdU-incorporated cells in trunk crest cell cultures was similar in the presence and absence of RA. The results suggest that in the early stages of in vitro development, RA has a larger impact on proliferation and/or survival of cranial neural crest cells than of trunk neural crest cells. Moreover, the data indicate that RA significantly affects melanogenesis and the differentiation of serotonergic neurons in mouse neural crest cell cultures from both axial levels. Whereas melanogenesis was suppressed by RA treatment, serotonergic neurogenesis was promoted. Double-labelling experiments with antibodies against BrdU and serotonin (5-HT) indicated that RA selectively promoted proliferation of these neurons at a later stage of in vitro development. Furthermore, RA acted upon both committed cells and multipotent cells, Based on the results, we conclude that RA plays multiple critical roles in mouse neural crest cell development.