THE FUNCTION OF THE ENDOTHELIN FAMILY IN NEURAL CREST DEVELOPMENT: AN IN VITRO SY

小鼠致死性斑点突变体（ls）编码血管活性肽内皮素3 (EDN3)，斑纹位点(s)编码其受体内皮素受体B （EDNRB）。这些基因突变纯合的小鼠表现出神经节巨克隆和白色斑点，这是由于在结肠和皮肤的受影响部分缺乏神经嵴衍生的肠神经节和黑素细胞。为了了解这些基因在黑素细胞发育中的作用，我们正在使用原位杂交来确定这些基因在突变和正常胚胎中相对于其他神经嵴标记的表达时间。我们已经证明，这两个基因在小鼠胚胎发育过程中在神经嵴衍生细胞中表达得非常早，并且在各种神经嵴突变体中表达模式发生改变。我们已经确定外源性添加EDN3改变促进黑色素细胞在体外系统的增殖和分化，我们已经开发。我们发现EDN3可以成功地取代TPA在这个系统中，并导致黑素细胞增加50倍。我们发现，在相关家族成员（EDN1和EDN2）中，只有EDN1可以替代EDN3。我们还表明，EDN3的作用依赖于其受体EDNRB在神经嵴细胞上的存在，通过使用化学抑制剂。-生物技术研究、癌症研究、消化系统疾病、基因定位（非人类）、神经科学、儿科研究；

The mouse mutant lethal spotting (ls) encodes the vasoactive peptide endothelin 3 (EDN3) and the piebald (s) locus encodes its receptor, endothelin receptor B (EDNRB). Mice homozygous for mutations in either of these genes exhibit aganglionic megaclon and a white spotted coat due to the lack of neural-crest derived enteric ganglia and melanocytes in affected portions of the colon and skin. In order to understand the role of these genes in melanocyte development, we are using in situ hybridization to determine the time that these genes are expressed relative to other neural crest markers in mutant and normal embryos. We have shown that both genes are expressed very early during mouse embryonic development in neural crest derived cells and expression patterns are altered in various neural crest mutants. We have determined that exogenously added EDN3 alters promotes proliferation and differentiation of melanocytes in an in vitro system that we have developed. We have found that EDN3 can successfully replace TPA in this system and results in a 50 fold increase in melanocytes. We have found that of the related family members (EDN1 and EDN2) only EDN1can substitute for EDN3 in this assay. We have also shown that the effect of EDN3 is dependent upon the presence of its receptor EDNRB on neural crest cells by using chemical inhibitors. - biotechnology research, cancer research, digestive diseases, gene mapping(non-human), neuroscience, pediatric research,