MOLECULAR GENETICS OF PANCREAS DEVELOPMENT

这项工作的长期目标是了解胰腺癌的细胞和分子机制 在脊椎动物胚胎发生过程中的发育。在过去的2.5年里，我们进行了筛选。 使用转基因斑马鱼系调节胰腺发育的基因， 内胚层和内胚层衍生的器官。我们还开发了全血免疫染色 技术，并利用新的转基因株系来表征这些突变体。重要的是，我们执行了 对斑马鱼野生型胰腺发育的详细分析（Field等人，2003 b），一项研究， 促进了斑马鱼系统的使用，以鉴定和/或测试与该过程有关的基因。 具体来说，我们发现斑马鱼胰腺是由两个芽融合而成的，一个是后部芽 它只包含胰岛细胞和一个产生外分泌细胞和胰腺细胞的前部细胞。 管道。胰管又产生新的胰岛细胞。这种内分泌和 外分泌谱系应有助于他们的调查。 我们建议继续进行研究，具体目标如下。 1)分析我们筛选出的胰腺突变体。 2)进一步表征6个突变体，包括详细的表型分析和受影响的分离。 基因 3)分析周围组织在胰腺发育和β细胞分化中的作用。我们 将详细分析最初的胰岛细胞（形成后芽的细胞）的出现， 目标是识别调节这一过程的信号。这些信息将有助于设计细胞 将干细胞分化为β细胞的培养方案。我们还将测试信号的假设， 从侧板中胚层控制前胰芽的发育，以及 新的胰岛细胞从胰管中出现。 4)继续开发试剂和技术，以方便使用的斑马鱼作为快速筛选 基因调控胰腺发育和β细胞分化的工具。

The long-term goals of this work are to understand cellular and molecular mechanisms of pancreas development during vertebrate embryogenesis. Over the past 2.5 years, we have conducted ascreen.for genes regulating pancreas development using a transgenic zebrafish line expressing GFP in the endoderm and endoderm-derived organs. We have also developed wholemount immunostaining techniques and utilized novel transgenic lines to characterize these mutants. Importantly, we carried out a detailed analysis of wild-type pancreas development in zebrafish (Field et al., 2003b), a study that has facilitated the use of the zebrafish system to identify and/or test genes implicated in this process. Specifically, we found that the zebrafish pancreas is formed by the fusion of two buds, a posterior one that contains only islet cells and an anterior one that gives rise to the exocrine cells and the pancreatic duct. The pancreatic duct in turn gives rise to new islet cells. This initial separation of endocrine and exocrine lineages should facilitate their investigation. We propose to continue our studies with the following specific aims. 1) Analyze the pancreas mutants identified and recovered from our screen. 2) Further characterize 6 mutants including detailed phenotypic analysis and isolation of the affected gene. 3) Analyze the role of the surrounding tissues in pancreas development and beta cell differentiation. We will analyze in detail the emergence of the initial islet cells (those that form the posterior bud) with the goals of identifying the signals that regulate this process. This information will be useful for designing cell culture protocols to differentiate stem cells into beta cells. We will also test the hypotheses that signals from the lateral plate mesoderm control the development of the anterior pancreatic bud, as well as the emergence of new islet cells from the pancreatic duct. 4) Continue to develop reagents and techniques to facilitate the use of the zebrafish as a rapid screening tool for genes regulating pancreas development and beta cell differentiation.