Oviductal regulation of gamete interaction and reproductive function

配子相互作用和生殖功能的输卵管调节

• 批准号: RGPIN-2015-04943
• 负责人: Kan, Frederick
• 金额: $ 1.75万
• 依托单位: Queen's University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=679914
• 关键词: Oviductal; regulation; gamete; interaction; reproductive

The mammalian oviduct undergoes physiological and biochemical modifications during late follicular development and estrus. These hormonally regulated changes optimize the oviductal microenvironment for fertilization and early embryonic development. A major glycoprotein secreted by oviductal cells, known as oviductin, is conserved across a number of mammalian species. In the hamster model used in our laboratory, oviductin is an O-glycosylated protein that undergoes cyclic variation of glycosylation during the estrous cycle. We have published extensively on the biosynthesis, secretion, localization and biochemical properties of hamster oviductin (HamOVGP1). During the tenure of my last NSERC DG, we demonstrated, for the first time, the ability of HamOVGP1 to enhance capacitation by potentiating tyrosine phosphorylation of a subset of sperm proteins known to play specific roles in sperm function. Our results further support an important role for oviductin in the oviductal milieu during the early events of fertilization. Recently, we have successfully produced recombinant hamster oviductin (rHamOVGP1) in HEK293 cells. Our recent work supported by NSERC reaffirms our laboratory in the forefront and as an authority in the field of mammalian oviductin. Based on our previous studies showing localization of oviductin to the cell surface of a subpopulation of epithelial cells of the hamster endometrium, we hypothesize that a putative oviductin receptor is present at the cell surface of certain uterine cells and that this receptor may play an important role in uterine receptivity. We plan 1) to examine the role of glycocomponents of rHamOVGP1 in enhancing sperm capacitation through enhancement of tyrosine phosphorylation of sperm proteins, and to assess the effect of glycan and glycopeptide derivatives of rHamOVGP1 in sperm-egg binding and fertilization; 2) to determine the mechanism of how rHamOVGP1 mediates signal transduction pathways that regulate protein tyrosine phosphorylation, and 3) to identify and characterize hamster oviductin receptor and to produce recombinant receptor for testing its possible role in uterine receptivity. To date, the mechanism of oviductin regulating the enhancement of capacitation and sperm-egg binding remains an enigma. Using the rHamOVGP1 that we successfully produced during the tenure of my last NSERC DG, it is now possible to unravel the mechanism of oviductin that regulates some of the key events that lead to successful fertilization and the possible role of oviductin in early implantation. It is anticipated that results obtained from this study will lead to the development of a suitable "drug cocktail" therapy utilizing oviductin and its glycan derivatives for improving the insemination procedures involving farm animals in the Canadian agriculture industry as well as IVF procedures involving rodents in research laboratories.***

哺乳动物输卵管在卵泡发育后期和发情期经历生理和生化变化。这些卵巢调控的变化优化了受精和早期胚胎发育的输卵管微环境。输卵管细胞分泌的一种主要糖蛋白，称为输卵管蛋白，在许多哺乳动物物种中是保守的。在我们实验室使用的仓鼠模型中，输卵管素是一种O-糖基化蛋白，在发情周期中经历糖基化的周期性变化。我们已经发表了大量关于仓鼠输卵管蛋白（HamOVGP 1）的生物合成，分泌，定位和生化特性。在我的最后一个NSERC DG任期内，我们首次证明了HamOVGP 1通过增强已知在精子功能中发挥特定作用的精子蛋白亚组的酪氨酸磷酸化来增强获能的能力。我们的研究结果进一步支持了输卵管蛋白在受精早期的输卵管环境中的重要作用。最近，我们已经成功地在HEK 293细胞中产生重组仓鼠输卵管蛋白（rHamOVGP 1）。我们最近的工作得到了NSERC的支持，这再次证明了我们的实验室在哺乳动物输卵管蛋白领域的前沿和权威地位。基于我们以前的研究显示本地化的输卵管素的细胞表面的一个亚群的上皮细胞的仓鼠子宫内膜，我们假设一个假定的输卵管素受体存在于某些子宫细胞的细胞表面，这种受体可能在子宫容受性中发挥重要作用。我们计划：1）研究rHamOVGP 1的糖组分通过增强精子蛋白的酪氨酸磷酸化而增强精子获能的作用，并评估rHamOVGP 1的聚糖和糖肽衍生物在精卵结合和受精中的作用; 2）确定rHamOVGP 1如何介导调节蛋白酪氨酸磷酸化的信号转导途径的机制，3）仓鼠输卵管素受体的鉴定和鉴定，以及重组受体的制备，以研究其在子宫容受性中的可能作用。迄今为止，输卵管素调节获能和精卵结合增强的机制仍是一个谜。使用我们在我的最后一个NSERC DG任期内成功生产的rHamOVGP 1，现在可以解开输卵管蛋白调节一些导致成功受精的关键事件的机制，以及输卵管蛋白在早期着床中的可能作用。预计从本研究中获得的结果将导致开发一种合适的“药物鸡尾酒”疗法，利用输卵管素及其聚糖衍生物改善加拿大农业中涉及农场动物的授精程序以及研究实验室中涉及啮齿动物的IVF程序。