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.

人类有数百种颅面疾病，其中腭裂最为常见。目标是 该提案旨在阐明腭发育背后的信号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来研究腭发育。强大的基因和 斑马鱼的细胞技术可用于研究基因功能以及细胞和组织 信号相互作用。此外，简化的腭骨骼由更少的腭神经嵴组成 斑马鱼胚胎的光学清晰度使其成为分析细胞的理想选择 腭发育过程中发生的行为。我建议检查互惠信号假说的预测， 其中信号从神经嵴到口腔外胚层，然后从口腔外胚层返回到神经嵴 诱导腭发育，并通过细胞嵌入引起上腭伸长。具体目标 1,1 检查候选基因对神经嵴衍生信号和口腔外胚层反应基因的作用， 在口腔外胚层中开启。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嵴的 FgflO 和 Bmp4 信号传导在口腔中开启pitx2的模型 外胚层，反过来又促进腭发育。在具体目标 2,1 中分析来自口头的相互信号 外胚层至神经嵴。我使用功能丧失、成像和遗传镶嵌分析以及 构建诱导型转基因斑马鱼品系以测试 Pdgf 和 Eph/ephrin 信号传导的预测 来自口腔外胚层的促进腭发育。在特定目标 3,1 中确定驱动的细胞行为 上颚伸长。我使用共焦延时分析以及小说的克隆和表征 斑马鱼上颚突变体来测试细胞嵌入驱动斑马鱼延伸的预测 味觉。我在这些研究过程中获得的结果将揭示遗传和细胞 腭裂的原因。此外，我建议分析的两个基因，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提供直接的见解 人类疾病的原因。