Characterization of genes required for border cell migration in Drosophila

果蝇边界细胞迁移所需基因的表征

基本信息

批准号：
8866193
负责人：
Lauren M. Anllo
金额：
$ 4.31万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8866193
关键词：
Abnormal Cell Actins Address Adhesions Affect Animals Anterior Antibodies Architecture Behavior Behavioral Biological Assay Biological Models Cancer Biology Cell Adhesion Cell Culture Techniques Cell Polarity Cells Complex Culture Media Cyclic AMP Cyclic AMP-Dependent Protein Kinases Defect Destinations Development Disease Drosophila genus E-Cadherin Epithelial Cells Epithelium Exhibits Fertility Follicle Stimulating Hormone G-Protein-Coupled Receptors Gene Expression Genes Genomics Health Homologous Gene Human Human Development Image Immigration LH Receptors Life Ligands Link Location Malignant Neoplasms Mammalian Cell Maps Membrane Migration Assay Modeling Molecular Morphology Motion Mutation Neoplasm Metastasis Nervous system structure Nurses Oocytes Oogenesis Organ Organism Ovary Pathway interactions Pattern Phenotype Play Process Proteins RNA Interference Regulation Reporter Rickets Role Running Signal Transduction Speed System Testing Time Traction Wing Work Wound Healing analog beta catenin bursicon cancer cell cell motility egg epithelial to mesenchymal transition experience human disease insight malignant breast neoplasm migration monolayer mutant nervous system development novel prevent receptor research study stem cell niche success tissue culture wound

项目摘要

DESCRIPTION (provided by applicant): During normal development of an organism, cells migrate from their original location to form specific organs, to populate stem cell niches and to form the complex nervous system. Abnormal cell migration is also observed in invasive cancer situations. In order to reveal the underlying mechanisms of cell migration, model systems such as the Drosophila ovary have been important in the past. Nevertheless, many questions about the regulation of cell migration are still unresolved, and this proposal addresses the role of a novel factor, Rickets, that we have found to be important for normal border cell migration in the Drosophila egg chamber. Rickets encodes a G-protein coupled receptor, and we have observed that border cells mutant for rk often fail to reach their destination at the anterior end of the oocyte. This proposal aims to investigate the underlying molecular causes of the migration defects and to understand the pathway in which Rk acts to promote cell migration. In Aim 1, I will identify the cellular architecture that is altered in the mutant cells. This will involve testng the level and distribution of important cell adhesion regulators, such as E-cadherin and Beta-Catenin, and polarity markers such as PAR-1 and PAR-3 in the mutant cells. I will also test whether Rk affects JAK/STAT signaling. In Aim 2, I will perform live imaging approaches to determine precisely what aspect of cell migration is altered in the mutant cells. I will focus on te transition of the border cells from epithelial to migratory morphology, on the detachment of the cells from the epithelium, on the speed and actual motion of the cells during migration (tumbling versus running) and on the extension of actin rich protrusions from the migrating cells. I will als test whether PKA and cAMP are acting downstream of Rk in the border cells. In Aim 3, I will test whether the effects or Rk on migration can also be observed in mammalian tissue culture. For this I will use an established system of breast cancer-derived cells that can be manipulated by RNAi approaches. A previously described scratch assay will allow me to determine whether mammalian cells with knockdown levels of Rk exhibit the same defects in migration as the mutant border cells in Drosophila. The proposed work will therefore define a new input into cell migration in Drosophila, and provide novel insights into this important developmental mechanism that are relevant to human development and disease.

描述（由申请人提供）：在生物体的正常发育过程中，细胞从其原始位置迁移以形成特定的器官，填充干细胞生态位并形成复杂的神经系统。在侵袭性癌症情况下也观察到异常细胞迁移。为了揭示细胞迁移的潜在机制，果蝇卵巢等模型系统在过去一直很重要。然而，有关细胞迁移调节的许多问题仍未得到解决，该提案解决了一种新因子“佝偻病”的作用，我们发现它对于果蝇卵室中正常边界细胞迁移很重要。佝偻病编码一种 G 蛋白偶联受体，我们观察到 rk 突变的边缘细胞常常无法到达卵母细胞前端的目的地。该提案旨在研究迁移缺陷的根本分子原因，并了解 Rk 促进细胞迁移的途径。在目标 1 中，我将识别突变细胞中发生改变的细胞结构。这将涉及测试突变细胞中重要的细胞粘附调节剂（例如 E-钙粘蛋白和 Beta-Catenin）以及极性标记（例如 PAR-1 和 PAR-3）的水平和分布。我还将测试 Rk 是否影响 JAK/STAT 信号传导。在目标 2 中，我将采用实时成像方法来准确确定突变细胞中细胞迁移的哪些方面发生了改变。我将重点关注边界细胞从上皮形态到迁移形态的转变、细胞与上皮的分离、细胞在迁移过程中的速度和实际运动（翻滚与奔跑）以及富含肌动蛋白的突起的延伸来自迁移的细胞。我还将测试 PKA 和 cAMP 是否在边界细胞中的 Rk 下游发挥作用。在目标 3 中，我将测试 Rk 对迁移的影响是否也可以在哺乳动物组织培养中观察到。为此，我将使用一个已建立的乳腺癌衍生细胞系统，该系统可以通过 RNAi 方法进行操作。先前描述的划痕测定将允许我确定具有 Rk 敲低水平的哺乳动物细胞是否表现出与果蝇中突变边界细胞相同的迁移缺陷。因此，拟议的工作将定义果蝇细胞迁移的新输入，并为与人类发育和疾病相关的这一重要发育机制提供新的见解。