Organogenesis of the Pharynx in C Elegans

描述（由申请人提供）：本提案中描述的实验将解决胚胎中塑造肠道器官发生的调节过程。我的实验室对前肠或咽的器官发生有着长期的兴趣，特别关注其转录调控过程。C.线虫是唯一适合这种分析，因为它提供了巨大的实验优势：透明度，单细胞分辨率，遗传学，快速转基因方法，短代时间，易于生物化学生长，简单的解剖学，强大的RNAi和日益强大的基因组学。在目标1中，我们将探索PHA-4与其靶基因的关联，以测试细胞命运特化的模型，并界定体内PHA-4结合的调节。我们将使用染色质免疫沉淀（ChIP）和体内成像来确定在不同时间和不同遗传背景下与PHA-4结合的基因。在目标2中，我们将测试这样的假设，即母亲的环境感知被传达给她的后代，这反过来又调节消化道发育，生长和/或生理。我们将分析咽部发育的野生型与PHA-4突变体，已妥协的化学感觉在特定的细胞或在特定的阶段。我们对本提案中描述的技术拥有丰富的经验，我们将与该领域的专家合作开发任何新技术。考虑到蠕虫和哺乳动物之间的保守性，我们发现的过程可能与理解人类器官发生和影响器官发育或功能的疾病（如癌症，糖尿病和代谢综合征）有关。

DESCRIPTION (provided by applicant): The experiments described in this proposal will address the regulatory processes that sculpt gut organogenesis in the embryo. My lab has a longstanding interest in organogenesis of the foregut, or pharynx, with a particular focus on its transcriptional regulatory processes. C. elegans is uniquely suited for this analysis because it offers tremendous experimental advantages: transparency, single-cell resolution, genetics, rapid transgene methodology, short generation time, ease of growth for biochemistry, simple anatomy, robust RNAi and increasingly powerful genomics. In Aim 1 we will explore PHA-4 association with its target genes to test models of cell-fate specification and to delimit regulation of PHA-4 binding in vivo. We will use Chromatin Immunoprecipitation (ChIP) and in vivo imaging to define genes bound by PHA-4 at different times and in different genetic backgrounds. In Aim 2, we will test the hypothesis that environmental sensing by the mother is communicated to her progeny, which in turn modulates digestive tract development, growth and/or physiology. We will analyze pharyngeal development in wild-type vs. pha-4 mutants that have been compromised for chemosensation in particular cells or at specific stages. We have extensive experience with the techniques described in this proposal, and we will collaborate with experts in the field for any new technology. Given the conservation between worms and mammals, the processes we uncover will likely be relevant for understanding human organogenesis and diseases that affect organ development or function such as cancer, diabetes and metabolic syndrome.