Prostaglandins and female reproductive function

女性生殖内分泌学是我实验室进行的一般研究项目。反刍动物（牛）怀孕的识别和建立是最重要的组成部分，已经由NSERC GDP资助了26年。前列腺素是女性生殖功能的关键调节因子，特别是参与子宫和卵巢之间的交流，以调节发情周期的持续时间。我们已经提出PGE2和PGF2a而不是单独的PGF2a决定黄体溶解或维持妊娠时的CL。在目前的资助周期中，我们将确认我们新发现的PGF合成酶AKR1B1的作用，它与PGE合成酶的调节和反馈相互作用，以实现PGE/PGF比例的严格控制。我们将通过极化子宫内膜上皮细胞培养研究PG通过2类转运蛋白SLCO和ABCC蛋白在细胞膜和组织中的转运及其在妊娠识别中的作用。虽然原生PGE2和PGF2a将通过我们验证的EIA方法进行分析，但PGDH局部与全身前列腺素失活的分析将依赖于新开发的LC/MS系统，以同时分析细胞培养基、血液和子宫组织中多达12种PG代谢物的存在。通过细胞培养获得的结果将在体内条件下使用子宫内膜外植体进行验证。在我们计划的研究中使用的验证技术包括qRT-PCR，免疫组化，Western blotting，极化上皮细胞在膜插入物上的培养，siRNA和选定基因的瞬时转染。

Endocrinology of female reproduction is the general research program conducted in my laboratory. Recognition and establishment of pregnancy in ruminants (cattle) is the most important component and has been funded by NSERC GDP for 26 years. Prostaglandins are key regulators of female reproductive function and particularly involved in the communication between the uterus and the ovary to regulate the duration of the estrous cycle. We have proposed that PGE2 and PGF2a rather than PGF2a alone determines luteolysis or maintenance of the CL at the time of recognition of pregnancy. In the present grant cycle, we will confirm the role of our newly identified PGF synthase AKR1B1, its regulation and feed-back interaction with PGE synthases to effect a tight control of the PGE/PGF ratio. We will study the transit of PGs across cell membranes and tissues through the 2 class of PG transporters SLCO and ABCC proteins using polarized endometrial epithelial cell cultures and its involvement during recognition of pregnancy. While native PGE2 and PGF2a will be assayed by our validated EIA assay, analysis of local vs. systemic inactivation of prostaglandins by PGDH will rely on a newly developed LC/MS system to analyze simultaneously the presence of up to 12 PG metabolites in cell culture medium, blood and uterine tissues. The results obtained with cell cultures will be validated using endometrial explants conditioned in vivo. The validated techniques used in our planned research include qRT-PCR, IHC, Western blotting, culture of polarized epithelial cells on membrane inserts, siRNA, and transient transfection of selected genes.