Neural Crest Cell Survival and Migration in Zebrafish

描述(由申请人提供) 这项建议旨在为应聘者提供为期1-2年的实验室指导研究，A.T.Look博士(Dana Farber)是使用斑马鱼作为研究周围交感神经系统和神经母细胞瘤发育的模型的先驱。这项提议还旨在支持候选人在神经生物学/肿瘤学系新教师任职的头3年内作为独立调查员。本应用的目的是以斑马鱼为模型，剖析Foxd3介导的神经脊细胞存活和迁移的分子组成，并确定这一途径是否促进体内神经脊来源的肿瘤的转移。斑马鱼与人类基因组密切相关，其保守的调节组织和器官发育的分子途径，为进行此类研究提供了一个强大的工具。这位候选人已经鉴定了一个斑马鱼突变体Foxd3，该突变体具有特定的神经脊细胞存活和迁移缺陷，并已确定Snail b转录因子是Foxd3功能的关键调节因子。这一应用的基本假设是，对Foxd3通路的了解将提供对神经脊细胞如何在外来环境中迁移和存活的理解，这是神经脊来源的肿瘤转移所必需的。在目标1中，遗传上位论和生物化学将确定Foxd3是否直接激活神经脊细胞中Snail b的表达，以及Snail b是否分别通过抑制有希望的候选基因PUMA和E-钙粘素的表达来抑制细胞凋亡和细胞迁移。在目标2中，将通过基因芯片分析来自Foxd3突变体和野生型同胞的GFP标记的神经脊细胞，以鉴定和分析Foxd3介导的神经脊细胞存活和迁移的其他下游靶点。在目标3中，Foxd3和蜗牛b将在发育中的神经脊细胞中过表达，以检测它们在已建立的斑马鱼神经脊肿瘤模型中促进转移的潜在作用。在正常发育期间，神经管中的细胞分离，激活生存程序，并迁移到胚胎的远处。这一过程类似于细胞在人类癌症转移过程中的行为。这项应用的目标是找到参与正常胚胎存活和迁移计划的基因，并确定这些基因在癌症转移过程中是否异常激活。

DESCRIPTION (provided by applicant) This proposal is designed to provide the candidate a 1-2 year period of mentored research in the laboratory of Dr. A. T. Look (at the Dana Farber), a pioneer in the use of zebrafish as a model to study development of the peripheral sympathetic nervous system and neuroblastoma. This proposal is also designed to support the candidate as an independent investigator during the first 3 years of a new faculty appointment in a neurobiology/oncology department. The goal of this application is to use the zebrafish as a model to dissect the molecular components of Foxd3-mediated neural crest cell survival and migration and to determine if this pathway promotes metastasis of neural crest-derived tumors in vivo. The zebrafish, with its close synteny to the human genome and its conserved molecular pathways regulating the development of tissues and organs, offers a powerful tool with which to conduct such research. The candidate has identified and characterized a zebrafish mutant Foxd3 line that has specific neural crest cell survival and migration defects and has identified the Snail b transcription factor as a critical mediator of Foxd3 function. The underlying hypothesis of this application is that knowledge of the Foxd3 pathway will provide an understanding of how neural crest cells migrate and survive in foreign environments, a requirement for the metastasis of neural crest-derived tumors. In Aim 1, genetic epistasis and biochemistry will determine if Foxd3 directly activates snail b expression in neural crest cells, and whether Snail b represses apoptosis and cell migration by repressing the expression of the promising candidates, puma and e-cadherin, respectively. In Aim 2, GFP-labeled neural crest cells from Foxd3 mutant and wild-type siblings will be analyzed by microarray to identify and analyze additional downstream targets of Foxd3-mediated neural crest cell survival and migration. In Aim 3, Foxd3 and snail b will be over-expressed in developing neural crest cells to examine their potential roles in promoting metastasis in established zebrafish neural crest tumor models. During normal development, cells from the neural tube detach, activate survival programs and migrate to distant regions of the embryo. This process is similar to the way cells behave during metastasis in human cancers. The goal of this application is to find the genes involved in the normal embryonic survival and migration programs and determine if these genes are aberrantly activated during cancer metastasis.