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Role of growth factors in catecholaminergic expression by neural crest cells: in vitro effects of transforming growth
1Department of Anatomy and Cell Biology, Columbia University, College of Physicians and Surgeons, New York, New York 10032.
Abstract:
The differentiation of neural crest cells into catecholaminergic neurons is dependent upon both intrinsic properties and signals from the embryonic microenvironment. In tissue culture, the development of catecholaminergic traits is dependent upon factors present in chick embryo extract (CEE). This dependency suggests that soluble growth factors affect catecholaminergic differentiation in vivo. We have studied the role of CEE-derived factors and the potentially related influence of characterized growth factors on catecholaminergic phenotypic expression in avian neural crest cells. In this report, we show that CEE-derived factors and transforming growth factor beta1 (TGF-beta 1) differentially influence catecholaminergic phenotypic expression as well as melanogenesis. TGF-beta 1 substituted for CEE-derived factors and supported the in vitro differentiation of tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH) immunoreactivities, as well as catecholamine biosynthesis and storage. Differentiation of catecholaminergic cells was dependent on factors present in 10% CEE during the first 1-4 days in culture suggesting an initial critical period for exposure. One day of initial exposure to either CEE-derived factors or TGF-beta 1 was sufficient to support the subsequent expression of catecholaminergic phenotypic characteristics. The time course of responsiveness to TGF-beta 1 was different than for CEE-derived factors. Neural crest cells remain responsive to TGF-beta 1 for at least 5 days, which is past the critical period for CEE-derived factors. Bioassay of CEE shows that endogenous levels of TGF-beta are less than or equal to 0.5 ng/ml. Immunoprecipitation of TGF-beta from CEE or blockade by neutralizing antibodies did not result in a loss of catecholaminergic differentiation by neural crest cells. Although CEE supports melanogenesis under all of the growth conditions tested, TGF-beta 1 was found to be inhibitory.