REGULATION OF STEROID HORMONE RECEPTOR ACTIVITY

类固醇激素受体活性的调节

• 批准号: 3230526
• 负责人: THOMAS G. MULDOON
• 金额: $ 10.62万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-04-01 至 1988-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3230526
• 关键词: androgens; bromocriptine; calcium; cyclic GMP; electrofocusing; estradiol; estrogen inhibitor; hormone analog; hormone regulation /control mechanism; luteinizing hormone; mammary gland; pituitary gland; prolactin; pyridoxal phosphate; radiotracer; steroid hormone biosynthesis; ultracentrifugation; uterus disorder

Several aspects of the mechanism of estrogen action dealing with regulation of estrogen receptor (ER) number and functionality will be investigated, using uteri, liver, anterior pituitary and hypothalamus from rats and mammary tissue of mice. The overall dynamics of fluctuating distribution of receptors will be studied as a function of occupied and unoccupied ER species in the intracellular compartments, including microsomes, at intervals after administration of various steriod hormone stimuli. Effects of ATP and/or molybdate, protein/RNA synthesis inhibitors, and dose-related ER processing, will be assessed. LH release patterns will be determined over the course of these receptor dynamics studies. The ER acceptor capability of microsomes will be explored as a possible means of detecting differences in ER complexes "activated" in response to binding of antiestrogens, androgens and 4-mercuriestradiol (4ME). The effects of 4ME, pyridoxal-5' phosphate and estradiol on microsomal RNP particles and cytosol initiation factors will be examined for dose- and time-dependency. Differences in mouse mammary gland ER nature and steroidal specificity in response to estrogen treatment will be investigated in the presence of protease inhibitors and with an eye toward existence of an estrone receptor. Prolactin augmentation and suppression will be tested as modifiers of ER and progesterone receptor activity, using mammary tissue from MTV+ and MTV strains of mice. In continued study of LHRH-ER interplay in the anterior pituitary, isolated ER-rich gonadotropes will be used. Calcium and cGMP will be examined as mediators of the action, and the kinetics of responsiveness to LHRH agonist/antagonist will be re-evaluated in these cells in suspension or culture. LHRH receptor distribution and microsomal component involvement will be studied. ER complexes activated in various ways excluding ligand binding will be assessed as competitors of 3H-estradiol-receptor complex binding to DNA, to measure the contribution of the steroid molecule itself to the interaction. The activated complexes will be physicochemically compared to see whether they are discernibly different. Intercalating agents, antiestrogen-receptor complexes, androgen-receptor complexes and unlabeled estradiol-receptor complexes will be used as putative competitors for DNA binding. Various sources of nucleotide sequences, including fragments of cloned estrogen-inducible genes, will be examined for binding specificity. These studies are expected to contribute to our understanding of the ways in which hormone responsiveness is subject to acute or chronic changes in the cellular environment.

雌激素作用机制涉及调控的几个方面 将研究雌激素受体(ER)的数量和功能， 用大鼠子宫、肝脏、垂体前叶和下丘脑 小鼠的乳腺组织。波动分布的总体动态 受体的数量将作为占用和未占用内质网的函数进行研究 细胞内隔室中的物种，包括微生物体， 给予不同类固醇激素刺激后的间隔时间。效应 ATP和/或钼酸盐、蛋白质/RNA合成抑制剂和剂量相关 急诊室处理，将进行评估。将确定黄体生成素的释放模式 在这些受体动力学研究的过程中。ER的接受者 微粒子的能力将被探索为一种可能的检测手段 内质网复合体“激活”与受体结合的差异 抗雌激素、雄激素和4-汞雌二醇(4ME)。4ME的影响， 吡哆醛-5‘-磷酸和雌二醇对微粒体RNP颗粒的影响 将检查胞浆启动因子的剂量和时间依赖性。 小鼠乳腺ER性质和类固醇特异性的差异 对雌激素治疗的反应将在以下情况下进行调查 蛋白酶抑制剂，并着眼于雌酮的存在 受体。催乳素的增强和抑制将被测试为 雌激素受体和孕激素受体活性修饰剂，使用乳腺组织 来自MTV+和MTV品系的小鼠。在LHRH-ER相互作用的继续研究中 在垂体前叶，将使用分离的富含内质网的促性腺激素。 钙和cGMP将作为这一作用的中介物质进行检查，而 对LHRH激动剂/拮抗剂的反应动力学将重新评估 在悬浮或培养的这些细胞中。促性腺激素释放激素受体的分布和 我们将研究微粒体组分的参与。Er络合物被激活 以各种方式排除配体结合将被评估为竞争对手 ~3H-雌二醇受体复合体与DNA的结合，以测量其贡献 类固醇分子本身的相互作用。活化的络合物 将进行物理化学比较，看看它们是否可辨别 不一样。插层剂，抗雌激素受体复合体， 雄激素受体复合体和未标记的雌二醇受体复合体 被用作DNA结合的假定竞争对手。各种来源的 核苷酸序列，包括克隆的雌激素诱导的片段 基因，将被检查结合的特异性。这些研究是 希望有助于我们理解荷尔蒙 反应性受制于细胞的急性或慢性变化。 环境。