OVARIAN CATECHOLESTROGENS

该提案涉及儿茶酚雌激素的自分泌/旁分泌作用， 猪的卵泡 我们的数据表明，这些新的 化合物作为局部卵巢调节剂具有很大的潜力， 在卵巢卵泡中合成， 卵巢滤泡细胞 在预计的支持期间，我们将 旨在更详细和全面地了解这一假定的 监管系统在每一个几个点，可以决定他们的 冲击 儿茶酚雌激素的生物合成将进一步研究， 了解关键的生物合成酶的性质和调节 雌激素-2-羟化酶（E-2-H）。 为了解决这个问题，荷尔蒙 将在培养的颗粒层和卵泡膜中研究这种酶的调节 细胞和卵泡组织从促性腺激素处理秋千。 水平 儿茶酚雌激素本身也将在卵泡液中测定， 和培养基。 卵巢E-2-H及其mRNA的性质将是 用抗体和cDNA探针研究。 一旦得到验证，这些试剂将 导致对酶的生物合成及其调节的详细研究。 下一个是 我们将研究儿茶酚雌激素代谢的意义， 卵巢，因为这些类固醇的降解对其水平很重要， 行动上 对于这些研究，我们将通过儿茶酚-O- 甲基转移酶（COMT），其位置和激素控制。 我们还 将确定2-O-甲基-雌二醇的作用机制，主要 COMT的产品 最后，我们将确定细胞位点， 儿茶酚雌激素对卵巢颗粒细胞的作用机制。 在这些研究中，我们将评估儿茶酚雌激素对 活性和生物合成的酶的生物合成 通路 特别是，儿茶酚雌激素对环磷酸腺苷的影响 生成，并在调节HMG-CoA-还原酶和胆固醇侧- 将测定链断裂。 这些研究预计将建立 这些化合物在排卵前发育中的独特作用 毛囊

This proposal addresses autocrine/paracrine actions of catecholestrogens in the porcine ovarian follicle. Our data have shown that these novel compounds have substantial potential as local ovarian regulators since they are synthesized in the ovarian follicle and act to promote differentiation of ovarian follicular cells. In the projected period of support, we will aim at a more detailed and integrated understanding of this putative regulatory system at each of several points which may determine their impact. The biosynthesis of catecholestrogens will be further studied by understanding the nature and regulation of the key biosynthetic enzyme -- estrogen-2-hydroxylase (E-2-H). To approach this issue, the hormonal regulation of this enzyme will be studied in cultured granulosa and theca cells and in follicular tissue from gonadotropin-treated swing. Levels of the catecholestrogens themselves will also be determined in follicular flui and culture medium. The nature of the ovarian E-2-H and its mRNA will be studied with antibodies and cDNA probes. Once validated, such reagents wil lead to detailed studies of enzyme biosynthesis and its regulation. Next, we will study the significance of the metabolism of catecholestrogens in th ovary, since degradation of these steroids is important to their levels and action. For these studies, we will examine CE metabolism via catechol-O- methyltransferase (COMT), its location and hormonal control. Further, we will determine the mechanism of action of 2-O-methyl-estradiol, the chief product of COMT. Finally, we will determine the cellular locus and mechanism of action of the catecholestrogens on ovarian granulosa cells. In these studies, we will evaluate the effects of catecholestrogens on the activity and biosynthesis of the enzymes of the progestin biosynthetic pathway. In particular, effects of catecholestrogens on cyclic AMP generation, and in regulation of HMG-CoA-reductase and cholesterol side- chain cleavage will be determined. These studies are expected to establish a unique role for these compounds in the development of the preovulatory follicle.