ECM AND CELL MIGRATION IN MULLERIAN DUCT REGRESSION

苗勒管回归中的 ECM 和细胞迁移

• 批准号: 2203437
• 负责人: HARRIET B AUSTIN
• 金额: $ 6.94万
• 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 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2203437
• 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这种机制。 以鳄鱼为模型研究苗勒管回归 系统。作为胚胎输卵管的米勒管发育 在两性脊椎动物中，然后在雄性脊椎动物中由于 睾丸激素的作用，MIS(苗勒氏抑制物质)。 在哺乳动物中，退化的特征是 细胞外基质(ECM)；在哺乳动物、鸟类和鳄鱼中工作 提示它涉及导管的去分化和迁移 细胞转移到胚胎中的其他位置。拟议的研究将 调查ECM中的更改如何导致与以下各项相关的更改 成纤维细胞生长因子是否参与退行性变 这一过程。因此，这项研究有可能使 对当前对一些 两种正常组织形态发生的基本过程 以及恶意性。 计划进行三组实验。第一，导管的迁移 体内的细胞将得到证实，因为它只在体外观察到。 第二，特定的ECM分子的作用，如层粘连蛋白和 纤维连接蛋白将通过免疫组织化学进行检测，以确定 这些成分在回归前和回归过程中的变化。此外, 将ECM抗体应用于体外培养的导管，观察 阻断细胞与细胞外基质结合的作用。这项实验将提供 关于哪些ECM分子直接参与促进 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和促进细胞迁移，以及 它在降解后从基底膜中释放出来，其 可能作为上皮细胞转化的诱导剂和 将使用以下工具调查回归过程中的迁移 免疫组织化学以确定它出现的时间和地点。 抗成纤维细胞生长因子的抗体也将应用于体外培养的导管，以阻断其 效果。