Genetic and cellular analysis of germ cell transepithelial migration

生殖细胞跨上皮迁移的遗传和细胞分析

• 批准号: 7799133
• 负责人: JESSICA SEIFERT
• 金额: $ 5.17万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2010-06-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7799133
• 关键词: Behavior; Cadherins; Cancer Biology; Cells; Developmental Biology; Drosophila genus; Embryo; G-Protein-Coupled Receptors; Genetic; Germ Cells; Goals; Immunohistochemistry; Immunology; Lead; Malignant Neoplasms; Midgut; Mutation; Primary Neoplasm; Primordium; Process; Property; Protein Region; Recycling; Regulation; Role; Site; Structure; System; Testing; Tissues; Work; cancer cell; cell motility; in vivo; migration; named group; next generation; novel; tumor

DESCRIPTION (provided by applicant): Understanding the regulation of cell migration is essential to the fields of developmental biology, immunology and cancer biology. However we lack a fundamental understanding1 of the strategies employed by migrating cells in vivo. The long-term goal of this proposal is to use Drosophila germ cell migration as an in vivo system to study cell migration and invasive behavior. Tre1 is a G-protein coupled receptor (GPCR) that is required specifically for germ cells to migrate through the posterior midgut primordium. Our hypothesis is that Tre1 regulates germ cell transepithelial migration by polarizing germ cells and regulating the localization of DE-cadherin. To test this hypothesis we will first determine the extent of germ cell polarization by Tre1 using ultra structural analysis and immunohistochemistry in wild type and trel backgrounds. We will also determine if polarization is a direct consequence of restricted Tre1 localization and/or activation. Secondly, we will test whether Tre1 is responsible for directed transport, targeted recycling and/or targeted degradation of DE-cadherin, leading to restricted DE-cadherin localization. Lastly, we will perform structure-function analysis of DE-cadherin to identify regions of the protein responsible for localization. We will use this region as bait to identify downstream effectors of Tre1 that are responsible for DE-cadherin localization. Through this work we will determine the mechanism by which Tre1 regulates germ cell polarization and DE-cadherin localization, allowing us to characterize a novel role for GPCRs in invasive migration. Germ cells are a special population of cells that are responsible for producing the next generation and as such they have special properties, for instance they are able to migrate through the tissue from one site to another early in the embryo. During cancer, some cells from the primary tumor gain the ability to migrate away from the original tumor and form additional tumors at new sites. By studying the processes occurring in germ cells that allow them to migrate we will be able to uncover how mutations in cancer cells lead to their motile behavior.

描述（申请人提供）：了解细胞迁移的调控对发育生物学，免疫学和癌症生物学领域至关重要。然而，我们对细胞在体内迁移所采用的策略缺乏基本的了解。本课题的长期目标是利用果蝇生殖细胞迁移作为体内系统来研究细胞迁移和侵袭行为。Tre1是一种g蛋白偶联受体（GPCR），是生殖细胞通过后中肠原基迁移所必需的。我们的假设是Tre1通过极化生殖细胞和调节DE-cadherin的定位来调节生殖细胞经上皮迁移。为了验证这一假设，我们将首先使用超结构分析和免疫组织化学在野生型和trel背景下确定Tre1对生殖细胞极化的程度。我们还将确定极化是否是Tre1受限定位和/或激活的直接结果。其次，我们将测试Tre1是否负责DE-cadherin的定向运输、靶向回收和/或靶向降解，从而导致DE-cadherin定位受限。最后，我们将对de -钙粘蛋白进行结构-功能分析，以确定负责定位的蛋白质区域。我们将使用这个区域作为诱饵来鉴定Tre1下游的效应物，这些效应物负责de -钙粘蛋白的定位。通过这项工作，我们将确定Tre1调节生殖细胞极化和DE-cadherin定位的机制，使我们能够表征gpcr在侵袭性迁移中的新作用。生殖细胞是一种特殊的细胞群，负责产生下一代，因此它们具有特殊的特性，例如，它们能够在胚胎早期通过组织从一个位置迁移到另一个位置。在癌症期间，来自原发肿瘤的一些细胞获得了从原始肿瘤迁移并在新部位形成额外肿瘤的能力。通过研究生殖细胞中允许它们迁移的过程，我们将能够揭示癌细胞的突变是如何导致它们的运动行为的。