ZEBRAFISH NEUROGENESIS-- EMBRYOLOGY AND GENETICS

描述(申请人的摘要逐字复制)：本报告的目标 建议是了解神经系统后部是如何 (后脑和脊髓)最初在脊椎动物中确定，使用 斑马鱼作为一种范例。斑马鱼是一个很棒的模型，可以进行分析 神经系统发育的早期、关键阶段，由两者 胚胎学和遗传学分析。这项研究的一个组成部分是一组基因 我们是通过差减克隆分离出来的，它只在 原肠形成过程中胚胎的后部，此时神经模式具有 已经开始，但在神经分化发生之前。其中一些基因 分离的含有DNA结合基序，而有些是新的，这表明我们 丰富了调控基因。这个应用程序有三个部分。第一, 利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分析来确定诱导 我实验室里的斑马鱼。第二，诱导的因素 后部神经组织将使用斑马鱼突变体和显性突变体进行分析 负性蛋白质。第三，通过以下方式分离的基因子集的功能 我们将讲解减法。基因将被放置在斑马鱼的基因上 映射以确定它们是否对应于已知的突变。函数的增益 分析将询问新的nlz基因是如何调节后脑的神经模式的。 这个应用程序将定义细胞相互作用和所需的基因 后脑和脊髓发育正常。正常的大脑发育和 健康需要神经系统所有部分的整合，这表明 后部神经系统的缺陷可能导致或加剧 其他脑区功能障碍。这里将要研究的基因也可能会有所帮助。 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.