ACTION OF GONADOTROPINS AND EICOSANOIDS IN LUTEAL CELLS

促性腺激素和二十烷类化合物在黄体细胞中的作用机制是 很复杂，而且还没有完全理解。这些代理的操作可能不会 完全可以通过细胞表面受体的结合和激活来解释 腺苷环化酶。这些药物内化并与受体结合。 存在于一个或多个细胞内细胞器中。在我们的努力中 测定核促性腺激素和前列腺素的生物学意义 结合位点，我们已经成功地证明了 促性腺激素、前列腺素E和前列腺素F2a反应核苷三磷酸酶(NTPase)， 牛黄体中参与核质内信使核糖核酸转移的一种酶 核膜。这些发现提出了一些额外的问题， 这项拨款提案旨在回答这一问题。简而言之， 促性腺激素和前列腺素对离体黄体核mRNA转运的影响 促性腺激素和前列腺素反应的NTPase是否在其中起作用 利用cDNAs探针运输和鉴定转运蛋白 调查过了。PGS还与核膜结合并刺激NTPase 活动。但这一反应还没有得到详细的描述，它 将在目前的研究中完成。腺苷是否会放大 促性腺激素对环磷酸腺苷和孕酮的作用 生产中，还可以放大NTPase对hCG和PGs的反应 检查过了。基础NTPase活性及其对促性腺激素和PGs的反应 在黄体快速生长、结构和功能成熟的时期 妊娠成功挽救的黄体中的退化 将会被研究。黄体切片将与和 没有促性腺激素和前列腺素，核膜就会被分离出来 用于测量NTPase活性。牛黄体核中的NTPase 膜将被溶解和纯化，以回答该酶是否 是否能与促性腺激素和前列腺素结合，以及这些配体是否能刺激 纯化的酶。拟议的实验是我们的 最新的发现。这个项目提供了几个新的想法，可以 潜在地在分子领域贡献了新的概念 促性腺激素和前列腺素在黄体中的作用机制。这样的概念 不一定与细胞表面结合的重要性相冲突 促性腺激素和前列腺素的作用与配体的最终降解 和/或其在黄体细胞溶酶体中的受体。新的概念可以 补充或补充促性腺激素和前列腺素的作用机制 行动。

The mechanism of gonadotropins and eicosanoids action in luteal cells is complex and not fully understood. The action of these agents may not completely be explained by cell surface receptor binding and activation of adenyl cyclase. These agents internalize and associate with receptors present in one or more of the intracellular organelles. In our efforts to determine the biological significance of nuclear gonadotropin and PGs binding sites, we have succeeded in demonstrating the presence of gonadotropin, PGE and PGF2a responsive nucleoside triphosphatase (NTPase), an enzyme involved in nucleocytoplasmic transfer of mRNA, in bovine luteal nuclear membranes. These findings raise a number of additional questions, which this grant proposal aims to answer. Briefly, the effect of gonadotropins and PGs on mRNA transport from isolated luteal nuclei, whether gonadotropin and PGs responsive NTPase plays a role in this transport and identification of transported mRNA using cDNA probes will be investigated. PGs also bind to nuclear membranes and stimulate NTPase activity. But this response has not been characterized in detail and it will be done in the present studies. Whether adenosine, which amplifies gonadotropins action with respect to cyclic AMP and progesterone production, can also amplify NTPase response to hCG and PGs will be examined. Basal NTPase activity and its response to gonadotropins and PGs during periods of rapid luteal growth, structural and functional maturity and regression and in corpora lutea rescued by establishment of pregnancy will be studied. Slices of corpora lutea will be incubated with and without gonadotropins and PGs and then nuclear membranes will be isolated for measurement of NTPase activity. NTPase in bovine luteal nuclear membranes will be solubilized and purified to answer whether this enzyme can bind gonadotropins and PGs and whether these ligands can stimulate the purified enzyme. The proposed experiments are a logical extension of our most recent findings. This project offers several new ideas and can potentially contribute novel concepts in the area of the molecular mechanism of gonadotropin and PGs action in corpus luteum. Such concepts do not necessarily conflict with the importance of cell surface binding in the action of gonadotropins and PGs and eventual degradation of ligands and/or their receptors in lysosomes of luteal cells. The new concepts can be complimentary to or additional mechanisms of gonadotropin and PGs action.