ZEBRAFISH NEUROGENESIS-- EMBRYOLOGY AND GENETICS

描述（申请人的摘要逐字复制）：本申请的目的 我们的建议是了解神经系统的后部区域 （后脑和脊髓）最初在脊椎动物中确定，使用 斑马鱼作为一个范例。斑马鱼是一个很好的模型， 神经系统发育的早期关键阶段， 胚胎学和遗传学分析。这项研究的核心是一组基因， 我们通过消减克隆分离出来的， 在原肠胚形成期间，当神经模式已经 开始，但在神经分化发生之前。的一些基因 分离的含有DNA结合基序，而有些是新的，这表明我们 富含调节基因。该应用程序有三个部分。第一、 使用这些基因作为标记，负责后神经的组织 将使用开发的外植体测定法确定诱导， 在我的实验室里。第二，诱导所需的因素 将使用斑马鱼突变体和显性突变体分析后部神经组织。 阴性蛋白质第三，通过分离基因的子集的功能， 将处理减法。基因将被放置在斑马鱼的遗传 以确定它们是否对应于已知的突变。功能获得 分析将询问新的NLZ基因如何调节后部神经模式。 该应用程序将定义细胞相互作用和基因所需的 正常的后脑和脊髓发育正常的大脑发育和 健康需要整合神经系统的所有部分，这表明， 后神经系统的缺陷可能会导致或加剧 其他脑区的功能障碍。这里要研究的基因也可能有所帮助 明确神经管先天缺陷的遗传基础

DESCRIPTION (Applicant's abstract reproduced verbatim): The goals of this proposal are to understand how posterior regions of the nervous system (hindbrain and spinal cord) are initially determined in vertebrates, using the zebrafish as a paradigm. The zebrafish is a wonderful model, allowing analysis of the early, crucial stages of nervous system development, by both embryological and genetic analysis. Integral to this study is a set of genes that we isolated by subtractive cloning, and that is expressed exclusively in the posterior of the embryo during gastrulation, when neural patterning has begun but before neural differentiation has occurred. Some of the genes isolated contain DNA binding motifs, while some are novel, suggesting that we have enriched for regulatory genes. This application has three parts. First, using these genes as markers, the tissues responsible for posterior neural induction will be determined, using explant assays that were developed for zebrafish in my laboratory. Second, the factors required for induction of posterior neural tissue will be analyzed using zebrafish mutants and dominant negative proteins. Third, the function of a subset of genes isolated by subtraction will be addressed. Genes will be placed on the zebrafish genetic map to determine whether they correspond to known mutations. Gain of function assays will ask how the novel nlz gene modulated posterior neural patterning. This application will define the cell interactions and genes required for normal hindbrain and spinal cord development. Normal brain development and health require integration of all parts of the nervous system, suggesting that deficits in the posterior nervous system may contribute to or exacerbate malfunction of other brain regions. The genes to be studied here may also help define the genetic basis of neural tube birth defects.