ZEBRAFISH GENES INVOLVED IN EARLY EMBRYONIC DEVELOPMENT

为了了解人类遗传病的基础并开始 制定可以治疗人类发育异常的程序， 有必要建立适当的动物模型，并进行基本的 旨在揭示正常和正常的分子机制的研究 发育不正常。分子遗传学研究进展 黑腹鱼等无脊椎动物的胚胎发育 而线虫则得益于它们较短的世代周期。为了澄清 脊椎动物胚胎发育的分子机制类似于 分析是必需的。然而，对分子遗传学基础的探讨 脊椎动物的发育受到了漫长的世代的阻碍，而且 在哺乳动物的情况下，无法获得的胚胎(即在子宫发育中)。 在这里提出的研究中，早期的分子遗传学分析 脊椎动物的发育将以斑马鱼为模型 系统。相对于其他生物，这种生物在实验上有很多优势 哺乳动物系统，包括体型小、繁殖力强和自然界 光学透明胚胎的体外发育。此外，它是 可能产生和分析雌核发育和单倍体后代，并 产生稳定的生殖系转化体。可能涉及的遗传基因座 在早期胚胎发育中，将通过一种 针对内源性增强子样作反应的转基因标记 位于整合地点附近的控制元素。独一无二的 这些集成的“增强子探测器”所表现出的表达模式将 为标记基因的功能提供重要线索。这个 这些转基因导致插入突变的可能性 有趣的表型也将被评估。转基因本身 提供杂交和/或遗传标记，允许快速恢复 识别出的基因座上的侧翼序列。标记的轨迹将进一步 以南方和北方分析为特征，允许 识别可能在生物信息学中起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.