The Function of the Endothelin Family in Neural Crest Development: An In Vitro S

内皮素家族在神经嵴发育中的功能：体外研究

• 批准号: 6433633
• 负责人: William J Pavan
• 金额: --
• 依托单位: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6433633
• 关键词: cell differentiation; developmental genetics; endothelin; gene complementation; gene expression; gene mutation; genetic strain; hormone receptor; hormone regulation /control mechanism; in situ hybridization; laboratory mouse; melanocyte; neural plate /tube; polymerase chain reaction; tissue /cell culture

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,

小鼠突变体致死斑点（LS）编码血管活性肽内皮素3（EDN3），Piebald（S）基因座编码其受体内皮素受体B（EDNRB）。这些基因中的任何一个突变的纯合子均表现出Aganglionic Megaclon和白色斑点外套，这是由于缺乏神经克雷斯特衍生的肠神经神经节和黑色结肠和皮肤的黑素细胞。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相对于突变体和正常胚胎中其他神经rest标记的时间。我们已经表明，这两个基因在小鼠胚胎发育过程中很早就表达，在神经rest衍生的细胞中，表达模式在各种神经rest突变体中都会改变。我们已经确定，在我们开发的体外系统中，外源添加的EDN3改变了黑素细胞的增殖和分化。我们发现EDN3可以成功替代该系统中的TPA，并导致黑素细胞增加50倍。我们发现，在此测定中，相关家庭成员（EDN1和EDN2）仅替换EDN3。我们还表明，EDN3的作用取决于使用化学抑制剂的受体EDNRB对神经rest细胞的存在。 - 生物技术研究，癌症研究，消化疾病，基因图（非人类），神经科学，小儿研究，