ECM AND CELL MIGRATION IN MULLERIAN DUCT REGRESSION

苗勒管回归中的 ECM 和细胞迁移

• 批准号: 2392455
• 负责人: HARRIET B AUSTIN
• 金额: $ 7.83万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 1999-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2392455
• 关键词: Crocodilia; Mullerian duct inhibiting substance; alternatives to animals in research; cell migration; cell transformation; cell type; embryo /fetus cell /tissue; embryo /fetus tissue /cell culture; extracellular matrix proteins; fallopian tubes; fibroblast growth factor; fluorescent dye /probe; gel electrophoresis; histochemistry /cytochemistry; histogenesis; image processing; immunochemistry; integrins; statistics /biometry; western blottings

The long-range goal of the present study is to elucidate the mechanisms involved in Mullerian duct regression using crocodilians as a model system. The Mullerian ducts, which are the embryonic oviducts, develop in vertebrates of both sexes and then regress in the male due to the actions of the testicular hormone, MIS (Mullerian inhibiting substance). In mammals, regression is characterized by dramatic changes in the extracellular matrix (ECM); and work in mammals, birds and crocodilians suggests that it involves dedifferentiation and migration of the ductal cells to other locations in the embryo. The proposed research will investigate how alterations in the ECM cause the changes associated with regression and whether fibroblast growth factor ( FGF) contributes to this process. Therefore, this study has the potential to make a significant contribution to the current understanding of some of the fundamental processes that characterize both normal tissue morphogenesis as well as malignancy. Three sets of experiments are planned. First, migration of the ductal cells in vivo will be confirmed since it has only been observed in vitro. Second, the role of specific ECM molecules such as laminin and fibronectin will be examined using immunohistochemistry to identify changes in these components before and during regression. In addition, ECM antibodies will be applied to the ducts in vitro to observe the effect of blocking cell binding to the ECM. This experiment will provide information about which ECM molecules are directly involved in promoting cell migration. Third, because basic fibroblast growth factor (bFGF) has been shown to induce mesoderm as well as to promote cell migration, and it is released from basement membranes following their degradation, its possible function as the inducer of epithelial transformation and migration during regression will be investigated using immunohistochemistry to identify when and where it is present. Antibodies to FGF also will be applied to the ducts in vitro to block its effects.

本研究的长期目标是阐明其机制 参与使用鳄鱼作为模型的苗勒管回归 系统。 苗勒管（胚胎输卵管）发育 在两性脊椎动物中，然后由于以下原因在雄性中退化 睾丸激素 MIS（苗勒管抑制物质）的作用。 在哺乳动物中，退化的特点是身体的急剧变化 细胞外基质（ECM）；并在哺乳动物、鸟类和鳄鱼身上开展工作 表明它涉及导管的去分化和迁移 细胞转移到胚胎的其他位置。 拟议的研究将 研究 ECM 的改变如何导致相关的变化 回归以及成纤维细胞生长因子 (FGF) 是否有助于 这个过程。 因此，这项研究有潜力做出 对当前某些领域的理解做出了重大贡献 表征正常组织形态发生的基本过程 以及恶性肿瘤。 计划进行三组实验。 一、导管的迁移 由于仅在体外观察到，因此体内细胞将得到证实。 其次，特定ECM分子的作用，例如层粘连蛋白和 将使用免疫组织化学检查纤连蛋白以鉴定 这些成分在回归之前和期间的变化。 此外， ECM抗体将在体外应用于导管以观察 阻断细胞与 ECM 结合的作用。 本实验将提供 有关哪些 ECM 分子直接参与促进的信息 细胞迁移。 第三，因为碱性成纤维细胞生长因子（bFGF）具有 已被证明可以诱导中胚层并促进细胞迁移，并且 它在降解后从基底膜中释放出来，其 可能作为上皮转化诱导剂发挥作用 回归期间的迁移将使用以下方法进行调查 免疫组织化学以确定其存在的时间和地点。 FGF 抗体也将应用于体外导管以阻断其 影响。