Prostaglandins and the regulation of female reproductuve function

在过去的二十年里，NSERC的资金允许开发出几个来自不同动物物种子宫的细胞培养模型，以研究当地在胚胎-母体沟通过程中涉及的机制，特别是关于母体识别怀孕的机制。在不同物种中，子宫内膜细胞唯一最显著和最保守的特性是它们产生特定前列腺素(PG)的能力。性激素、滋养细胞干扰素(IFNtau)、催产素和血小板激活因子(PAF)可以调节单个细胞内PG的产生。在反刍动物中，由子宫内膜上皮细胞产生的PGF2f负责黄体的破坏和周期的恢复。另一方面，PGE2具有抗黄体溶解作用，具有不同的作用，有利于妊娠的建立。在确认怀孕时，胚胎必须调节子宫前列腺素的产生，以防止黄体溶解和回到新的周期。我们发现，在上皮细胞中，催产素刺激基础PGF2f的产生，而IFNtau则将其转化为PGE2。在过去的五年中，我们已经鉴定了牛子宫内膜的功能性PGF合成酶是醛糖还原酶AKR1B5，这是一种具有新功能的旧酶。我们还首次提出了新发现的PG转运蛋白PGT在PGs在生殖系统中的内分泌作用中的作用。最后，我们发现PG受体在子宫和黄体中的表达与生物合成和运输蛋白的表达模式非常匹配。我们的发现导致了一种新的范式，在母亲识别怀孕的过程中调节PG的行为。我们希望利用下一个授权期为PG在牛生殖系统中的作用提供一个新的综合观点。我们将利用不同生理条件下在体内收集的细胞培养和组织来研究这些系统。PGs的产生将通过EIA进行分析，由sn-6和omega-3脂肪酸产生的PG代谢物将通过GC/MS-MS进行分析。通过实时定量RT-PCR和Northern分析进行信使RNA的表达，通过Western blotting和IHC进行蛋白质水平的测定。

Over the last twenty years, NSERC funding allowed to develop several cell culture models derived from the uterus of different animal species to study the mechanisms involved locally during embryo-maternal communication especially with regard to maternal recognition of pregnancy. Across species, the single most remarkable and best conserved property of endometrial cells is their ability to produce specific prostaglandins (PG). PG production within a single cell can be modulated by sex steroids, trophoblastic interferons (IFNtau), oxytocin and platelet activating factor (PAF). In ruminants, PGF2fÑ produced by the endometrial epithelial cells is responsible for the destruction of the corpus luteum and the return to the cycle. On the other hand, PGE2 is antiluteolytic and plays different roles that favor the establishment of pregnancy. At the time of recognition of pregnancy, the embryo must regulate the production of uterine PGs to prevent luteolysis and the return to a new cycle. We have found that in epithelial cells basal PGF2fÑ production was stimulated by oxytocin but switched to PGE2 by IFNtau. Over the last five years, we have identified the functional PGFsynthase of the bovine endometrium as the aldose reductase AKR1B5, an old enzyme with a new function. We have also been the first to propose a role for the newly discovered PG transporter PGT in endocrine action of PGs in the reproductive system. Finally we have shown that the expression of PG receptors in the uterus and corpus luteum closely matches the pattern of expression of biosynthetic and transport proteins. Our findings lead to a new paradigm for the regulation of PG action during maternal recognition of pregnancy. We want to use the next granting period to provide a novel integrated view of PG action in the bovine reproductive system. We will study these systems using cell cultures and tissues collected in vivo under different physiological conditions.  The production of PGs will be analyzed by EIA and the full array of PG metabolites produced from sn-6 and omega-3 fatty acids by GC/MS-MS. Expression of messenger RNA will be done by real time RT-PCR and Northern analysis and protein levels by Western blotting and IHC.