Genetic Hierarchies and Cellular Behaviors during Zebrafish Palatogenesis

斑马鱼腭发育过程中的遗传层次和细胞行为

• 批准号: 7841071
• 负责人: JOHANN K EBERHART
• 金额: $ 0.85万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-02 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7841071
• 关键词: Back; Behavior; Biological Models; Brain; Candidate Disease Gene; Cartilage; Cell Proliferation; Cells; Cephalic; Cleft Palate; Cloning; Development; Disease; Dominant-Negative Mutation; Embryo; Ephrin-B1; Figs - dietary; Frontonasal Prominence; Gene Expression; Genes; Genetic; Goals; Human; Image; Individual; Life; Light; Mammals; Maps; Maxilla; Microscopy; Modeling; Molecular; Morphogenesis; Neural Crest; Neural Crest Cell; Optics; Palate; Physical condensation; Population; Role; Signal Transduction; Skeleton; Specific qualifier value; Techniques; Testing; Time; Tissues; Transgenic Organisms; Transplantation; Zebrafish; cell behavior; craniofacial; gene function; human disease; insight; intercalation; loss of function; mutant; novel; oral ectoderm; palatogenesis; progenitor; response; transcription factor

There are hundreds of craniofacial diseases in humans and cleft palate is common among these. The goal of this proposal is to elucidate the signaling interactions and cellular behaviors underlying palatogenesis. The zebrafish provides a useful model system in which to study palatal development. Powerful genetic and cellular techniques are available in the zebrafish for studying gene function as well as cell and tissue signaling interactions. Additionally, the simplified palatal skeleton, consisting of far fewer neural crest palate progenitors than in mammals, and the optic clarity of the zebrafish embryo makes it ideal for analyzing cell behaviors occurring in palatogenesis. I propose to examine predictions of a reciprocal signaling hypothesis, in which signals from neural crest to the oral ectoderm and then back from the oral ectoderm to neural crest induce palatogenesis, and cause elongation of the palate through cell intercalations. In Specific Aim 1,1 examine the role of candidate genes for neural crest-derived signals and oral ectoderm response genes, turned on in the oral ectoderm. I use loss-of-function, gene expression, imaging, and genetic mosaic analyses to test the model that FgflO and Bmp4 signaling from the neural crest turns on pitx2 in the oral ectoderm, which, in turn, promotes palatogenesis. In Specific Aim 2,1 analyze the reciprocal signal, from oral ectoderm to neural crest. I use loss-of-function, imaging, and genetic mosaic analyses as well as construction of inducible transgenic zebrafish lines to test the prediction that Pdgf and Eph/ephrin signaling from the oral ectoderm promotes palatogenesis. In Specific Aim 3,1 determine the cell behaviors that drive elongation of the palate. I use confocal time lapse analysis as well as cloning and characterization of novel zebrafish palate mutants to test the prediction that cell intercalations drive the extension of the zebrafish palate. The results I obtain during the course of these studies will shed light on the genetic and cellular causes of cleft palate. Additionally, two genes I propose to analyze, pitx2 and ephrin-B1, are known to be human craniofacial disease genes. Therefore, my analyses of these genes will provide direct insight into the cause of human disease.

人类中有数百种颅面疾病，在其中很常见。目标 该提议是为了阐明古质发生的信号传导相互作用和细胞行为。这 斑马鱼提供了一个有用的模型系统，可以在其中研究pa骨发展。强大的遗传和 斑马鱼中有细胞技术用于研究基因功能以及细胞和组织 信号交互。此外，简化的pa骨骨骼，包括较少的神经波峰 祖细胞比哺乳动物中的祖细胞，斑马鱼胚胎的视觉清晰度使其非常适合分析细胞 在古质发生中发生的行为。我建议检查相互信号假设的预测， 其中从神经rest到口服外胚层的信号，然后从口服外胚层到神经rest 诱导古氏生成，并通过细胞插入引起口感伸长。在特定的目标1,1 检查候选基因在神经rest衍生的信号和口服外胚层反应基因的作用，即 在口服外胚层中打开。我使用功能丧失，基因表达，成像和遗传镶嵌 分析以测试来自神经rest的FGFLO和BMP4信号传导在口服中的PITX2上的模型 外胚层又促进了古质发生。在特定目标中2,1分析互惠信号，来自口服 外胚层到神经rest。我使用功能丧失，成像和遗传镶嵌分析以及 构建可诱导的转基因斑马鱼线以测试PDGF和EPH/Ephrin信号传导的预测 从口服外胚层促进了古质发生。在特定目标中3,1确定驱动的细胞行为 口感的伸长。我使用共聚焦的时间流逝分析以及新颖的克隆和表征 斑马鱼的pa突变体以测试细胞插入驱动斑马鱼的延伸的预测 口感。在这些研究过程中，我获得的结果将阐明遗传和细胞 a裂的原因。另外，我提出的两个基因分析Pitx2和Ephrin-b1，已知是 人类颅面疾病基因。因此，我对这些基因的分析将直接洞悉 人类疾病的原因。