Exocrine Pancreas & Intestinal Epithelial Development

描述(由申请人提供)： 外分泌胰腺和肠在脊椎动物中起着重要的作用 生理学。对于这两个器官来说，细胞更新也有一个重要的动态平衡 角色。在肠道中，绒毛上皮细胞连续地 取而代之的是居住在 在每个绒毛周围的地窖里。在胰腺中，具有多能性 祖细胞位于导管上皮内，被认为是一种 分化的内分泌和腺泡细胞的来源，可以取代丢失的细胞 在特定的实验和病理条件下。身份识别 调节消化系统细胞谱系特征和分化的基因 器官干细胞前体细胞对与普通人类相关的研究很重要 胰腺癌和结肠癌等疾病。基因分析是一种 识别调控发育过程的基因的有效手段， 尤其是器官发生。在本应用程序中，我们概述了 家系特征与细胞分化的分子特征 脊椎动物斑马鱼发育中的外分泌胰腺和肠道 唯一适合这一目的的模型系统。外分泌胰腺和 斑马鱼仔鱼受精后5天肠道功能 符合标准诱变技术的阶段。此外，祖先和 两个器官的分化细胞在整个过程中都很容易被看到。 使用器官特异性标记物的发育。此应用程序的第一个目标是 对调节血统的基因进行系统的突变筛选 胰腺和肠道外分泌过程中的特异化和细胞分化 发展。利用原位和组织化学分析，我们将鉴定基因 调节外分泌胰腺的生长和分化，以及 肠的杯状细胞和内分泌细胞谱系的发育。 例如Screen的可行性在最近的一项试点研究中得到了证明。这个 第二个目标是在胰腺外分泌和 斑马鱼幼体的肠道，通过产生斑马鱼的胰腺和肠道 EST数据库。这一目标将与以下组织成员合作实现 斑马鱼基因组群落。第三个目标是绘制突变和EST的图谱 利用减数分裂和物理作图技术。这一目标将允许 特定目的恢复突变候选基因的鉴定 1，为基因敲除实验提供靶点数据库。第四次 这项应用的目的是探索新基因的功能。 具体目标2使用基于吗啡的基因敲除策略。

DESCRIPTION (Provided by Applicant): The exocrine pancreas and intestine play essential roles in vertebrate physiology. For both organs cell renewal also has an important homeostatic role. In the intestine, cells of the villus epithelium are continuously replaced by the differentiated progeny of stem cell progenitors that reside within the crypts surrounding each villus. In the pancreas, multipotent progenitors reside within the ductular epithelium and believed to serve as a source of differentiated endocrine and acinar cells that can replace cells lost under specific experimental and pathological conditions. Identification of the genes that regulate cell lineage specification and differentiation of digestive organ stem cell progenitors is important to research related to common human diseases such as cancers of the pancreas and colon. Genetic analysis is an efficient means to identify genes that regulate developmental processes, particularly organogenesis. In this application we outline a strategy for the molecular characterization of lineage specification and cell differentiation in the developing exocrine pancreas and intestine of the zebrafish, a vertebrate model system uniquely suited to this purpose. The exocrine pancreas and intestine of zebrafish larvae function at five days post‑fertilization, a stage amenable to standard mutagenesis techniques. Further, progenitors and differentiated cells of both organs are easily visualized throughout development using organ specific markers. The first Aim of this application is to perform a systematic mutagenesis screen for genes that regulate lineage specification and cell differentiation during exocrine pancreas and intestinal development. Using in situ and histochemical analysis we will identify genes that regulate growth and differentiation of the exocrine pancreas as well as development of the goblet and enteroendocrine cell lineages of the intestine. The feasibility of such as screen was proven in a recent pilot study. The second Aim is to identify molecular markers in the exocrine pancreas and intestine of larval zebrafjsh by generating a zebrafish pancreas and intestine EST database. This aim will be performed in collaboration with members of the zebrafish genome community. The third Aim is to map the mutations and ESTs using meiotic and physical mapping techniques. This aim will allow identification of candidate genes for the mutations recovered in Specific Aim 1, and provide a databank of targets for gene knockdown experiments. The fourth aim of this application is to explore the function of novel genes identified in Specific Aim 2 using a morpholino based gene knockdown strategy.