ROLE OF THE TETRASPANIN EMP2 IN EARLY REPRODUCTION

四跨膜蛋白 EMP2 在早期繁殖中的作用

• 批准号: 6536336
• 负责人: CARMEN J WILLIAMS
• 金额: $ 7.93万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-03 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6536336
• 关键词: animal breeding; biological signal transduction; egg /ovum; embryo implantation; embryogenesis; fertility; fertilization; gene expression; gene targeting; genetically modified animals; immunofluorescence technique; in vitro fertilization; laboratory mouse; membrane proteins; messenger RNA; northern blottings; polymerase chain reaction; protein localization; protein structure function; reproduction; uterus; vertebrate embryology; western blottings

The long term objective of this research is to further our understanding of the molecular mechanisms of fertilization, preimplantation embryo development , and implantation. Such studies are critical for the advancement of two clinically important areas: infertility and contraceptive development. To this end, we are using the mouse as a model system to study the reproductive biology that underlies these processes. We have recently identified on the surface of mouse eggs, embryos, and uterine epithelium, a novel tetraspanin protein called epithelial membrane protein 2 (EMP2). Tetraspanin proteins interact with cell surface integrins, and are emerging as critical regulators of many cellular processes, including cell adhesion, proliferation, and differentiation. EMP2 is known to interact with intergrin beta1, an intergrin thought to be critical for fertilization and possibly involved in implantation. The surface localization of EMP2, as well as known roles of previously identified tetraspanins, suggest a function for EMP2 in cellular interactions important for these early processes of reproduction. The first aim of this proposal is to characterize the expression pattern of EMP2 mRNA and protein in oocytes, embryos, and uterine tissue. This will be done using RT-PCR, immunofluorescence, and immunoblotting techniques. The second aim is to determine if inhibition of EMP2 expression and/or function results in inhibition of fertilization, preimplantation embryo development, or implantation. This will be done using both dominant negative and RNA interference approaches. In addition to improving our understanding of molecules relevant for the signal transduction pathways important in early reproduction, results of these studies may also have implications for clinical practice of assisted reproduction, as well as contraceptive development.

本研究的长期目标是进一步了解受精、着床前胚胎发育和着床的分子机制。这些研究对于不育和避孕发展这两个重要临床领域的进展至关重要。为此，我们正在使用小鼠作为模型系统来研究这些过程背后的生殖生物学。我们最近在小鼠卵子、胚胎和子宫上皮表面发现了一种新的四联蛋白，称为上皮膜蛋白2 （EMP2）。四联蛋白与细胞表面整合素相互作用，并成为许多细胞过程的关键调节因子，包括细胞粘附、增殖和分化。已知EMP2与间连蛋白β 1相互作用，而间连蛋白β 1被认为对受精至关重要，可能与着床有关。EMP2的表面定位，以及先前确定的四跨蛋白的已知作用，表明EMP2在细胞相互作用中具有重要的功能，对这些早期生殖过程很重要。本研究的第一个目的是确定EMP2 mRNA和蛋白在卵母细胞、胚胎和子宫组织中的表达模式。这将使用RT-PCR、免疫荧光和免疫印迹技术来完成。第二个目的是确定抑制EMP2表达和/或功能是否会抑制受精、着床前胚胎发育或着床。这将使用显性阴性和RNA干扰方法来完成。除了提高我们对早期生殖中重要的信号转导途径相关分子的理解外，这些研究的结果也可能对辅助生殖的临床实践以及避孕措施的开发具有启示意义。

项目成果

Epithelial membrane protein-2 regulates surface expression of alphavbeta3 integrin in the endometrium.

上皮膜蛋白 2 调节子宫内膜中 αvβ3 整合素的表面表达。

• DOI: 10.1016/j.ydbio.2005.09.003
• 发表时间: 2005
• 期刊: Developmental biology.
• 作者: Wadehra,Madhuri;Forbes,Ashley;Pushkarna,Natasha;Goodglick,Lee;Gordon,LynnK;Williams,CarmenJ;Braun,Jonathan
• 通讯作者: Braun,Jonathan