ZEBRAFISH GENES INVOLVED IN EARLY EMBRYONIC DEVELOPMENT

为了了解人类遗传疾病的基础，并开始 开发出治疗人类发育异常的程序， 有必要建立适当的动物模型， 研究旨在揭示正常和 异常发育分子遗传学描述的进展 无脊椎动物如D. melanogaster 和C.优雅线虫得益于其短的世代时间。阐明 脊椎动物胚胎发育的分子机制，类似于 需要分析。然而，对分子遗传基础的研究 脊椎动物的发育受到世代时间长的阻碍， 在哺乳动物的情况下，不可接近的胚胎（即，在子宫内发育）。 在这里提出的调查中，对早期的 脊椎动物的发展将开始使用斑马鱼作为模型 系统这种生物相对于 哺乳动物系统，包括小尺寸，繁殖力，和自然的， 光学透明胚胎的体外发育。此外还 可能产生和分析雌核发育和单倍体后代， 产生稳定的种系转化体。可能涉及的遗传位点 在早期胚胎发育中， 设计为响应内源性增强子样的转基因标记 控制位于整合位点附近的元件。独特的 由这些整合的“增强子检测器”显示的表达模式将 提供了重要的线索，以功能的标记基因。的 这些转基因导致插入突变的可能性， 还将评估感兴趣的表型。转基因本身 提供允许快速恢复的杂交和/或遗传标记 侧翼序列。标记的位点将进一步 以南方和北方分析为特征， 识别可能在以下方面发挥关键作用的单个基因： 发展

In order to understand the basis of human genetic disease and to begin to develop procedures by which human developmental anomalies can be treated, it is necessary to develop appropriate animal models and to perform basic research designed to uncover the molecular mechanisms of normal and abnormal development. Progress toward a molecular-genetic description of the embryonic development of invertebrate organisms such as D. melanogaster and C. elegans has been aided by their short generation times. To elucidate the molecular mechanisms by which vertebrate embryos develop, a similar analysis is required. However, inquiries into the molecular-genetic basis of vertebrate development have been hindered by long generation times, and in the case of mammals, inaccessible embryos (i.e., in utero development). In the investigations proposed here, a molecular-genetic analysis of early vertebrate development will be initiated using the zebrafish as a model system. This organism has many experimental advantages relative to mammalian systems, including small size, fecundity, and the natural in vitro development of optically transparent embryos. In addition, it is possible to produce and analyze gynogenetic and haploid progeny, and to produce stable germ-line transformants. Genetic loci likely to be involved in early embryonic development will be identified with the aid of a transgene marker designed to respond to endogenous enhancer-like controlling elements located near the site of integration. The unique expression patterns exhibited by these integrated "enhancer detectors" will provide important clues as to the function of the marked genes. The potential of these transgenes to result in insertion mutations with interesting phenotypes will also be evaluated. The transgenes themselves provide hybridization and/or genetic markers allowing the rapid recovery of flanking sequences at the identified locus. Marked loci will be further characterized by Southern and Northern analysis, allowing the identification of individual genes likely to play crucial roles in development.