NEURAL INFLUENCE ON HYPOPHYSEAL GONADAL FUNCTION

神经对垂体性腺功能的影响

• 批准号: 6592329
• 负责人: HAROLD G SPIES
• 金额: $ 11.11万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6592329
• 关键词: Mammalia; aging; animal tissue; clinical research; endocrine gland /system; hormones; lipids; nervous system; nucleic acids; proteins; reproductive system; structural biology; women's health

The preovulatory release of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) in women and rhesus monkeys is dependent upon enhanced secretion of ovarian estradiol-17? (E). This E-induced GnRH/LH surge is accompanied by changes in hypothalamic and brainstem cells that are identified as neuropeptide Y (NPY)-, norepinephrine (NE)-, acetylcholine (Ach)- and gamma aminobutyric acid (GABA)-containing neurons. Our research has focused on how E influences transcriptional-translational events associated with these peptide and transmitter systems and how these central neural events change with aging, especially the perimenopausal life-stage where E secretion declines. Two isoforms of the E receptor (ER ? and ?) have been reported. To study the tissue distribution of the ER isoforms, we cloned and sequenced two cDNA probes (126 bp and 291 bp) for rhesus monkey ER-?. These cDNA fragments were similar, but not identical, to human ER-? sequences. With pro bes for bo th ER-? and ER-?, we quantified the ER isoforms in several peripheral and central neural male and female tissues by reverse transcription-polymerase chain reaction (RT-PCR). Both the ER-? and ER-? mRNAs were present in male and female reproductive organs. Only ER-? was expressed in liver, frontal cortex, caudate nucleus, locus coeruleus and cerebellum. In some brain regions, i.e., the putamen, internal capsule, hippocampus and paraventricular hypothalamus, ER-? was expressed in the female, but not in the male. In macaque females 10-15 years of age mRNA expression for tyrosine hydroxylase (TH, enzyme for control of NE synthesis) in specific adrenergic neural populations (A1, A2, A6) are upregulated by E treatment. However, after E treatment in perimenopausal monkeys (> 22 years old), expression levels of TH mRNA is dampened in the A6 region of brainstem. We continue to study the relevance of this age- and E-related decline in TH mRNA response in brainstem neurons. We also cloned a 356 bp monkey choline acetyltransferase (CHAT, the enzyme for control of Ach synthesis) cDNA fragment that corresponds to the human CHAT N-1 type sequence 332 to 684. The 5= and 3= primer sequences were TAAGATGGCAGCAAAAACTCCC and ACCGATGACCAGCTGAGGTTT, respectively. To our knowledge, this partial DNA sequence of the rhesus monkey CHAT gene has not been documented previously. The MCHAT RNA probe that was generated from the clone hybridized with CHAT mRNA in several specific hypothalamic and brainstem areas in adult female macaque brain. Quantification of mRNA expression during various physiologic conditions will help establish the relevance of cholinergic neurons during ovulation and anovulation in primates. FUNDING NIH HD16631, HD18185 PUBLICATIONS Pau CY, Pau KYF, Spies HG. Putative estrogen receptor and a mRNA expression in male and female rhesus macaques. Mol Cell Endocrinol 146:59-68, 1998.

促性腺激素释放激素 (GnRH) 的排卵前释放 女性和恒河猴的黄体生成素 (LH) 具有依赖性 卵巢雌二醇17分泌增强？ (E)。 这种E诱发的 GnRH/LH 激增伴随着下丘脑和脑干的变化 被鉴定为神经肽 Y (NPY)-、去甲肾上腺素的细胞 (NE)-、乙酰胆碱 (Ach)- 和 γ 氨基丁酸 (GABA) 含有神经元。 我们的研究重点是E如何 影响与这些相关的转录翻译事件 肽和递质系统以及这些中枢神经事件如何 随着年龄的增长，尤其是围绝经期生命阶段，E 分泌减少。 E 受体的两种亚型（ER ? 和 ?） 被举报。 为了研究 ER 亚型的组织分布， 我们克隆并测序了恒河猴的两个 cDNA 探针（126 bp 和 291 bp） 猴子ER-？。 这些 cDNA 片段与 人类ER-？ 序列。 带有 ER- 的探头？ 和ER-？，我们 量化了几种外周和中枢神经中的 ER 亚型 通过逆转录聚合酶链检测男性和女性组织 反应（RT-PCR）。 两者都是ER-？ 和ER-？ mRNA 存在于 男性和女性的生殖器官。 只有ER-？ 被表达为 肝脏、额叶皮质、尾状核、蓝斑和 小脑。 在某些大脑区域，即壳核、内部 囊、海马和室旁下丘脑，ER-？ 曾是 在女性中表达，但在男性中不表达。 在雌性猕猴中 10-15 岁酪氨酸羟化酶（TH、 特定肾上腺素能神经中控制 NE 合成的酶） 人群（A1、A2、A6）通过 E 治疗上调。 然而， 围绝经期猴子（> 22岁）接受E治疗后， A6 区域 TH mRNA 的表达水平受到抑制 脑干。 我们将继续研究这个时代的相关性 脑干神经元中 TH mRNA 反应的 E 相关下降。 我们也 克隆了 356 bp 的猴胆碱乙酰转移酶（CHAT，该酶 Ach 合成的控制）对应于人类的 cDNA 片段 CHAT N-1 型序列 332 至 684。5= 和 3= 引物序列 分别是 TAAGATGGCAGCAAAAACTCCC 和 ACCGATGACCAGCTGAGGTTT。 据我们所知，恒河猴 CHAT 的部分 DNA 序列 基因以前没有被记录过。 MCHAT RNA 探针 由与 CHAT mRNA 杂交的克隆产生，在几个特定的 成年雌性猕猴大脑的下丘脑和脑干区域。 不同生理过程中 mRNA 表达的定量 条件将有助于建立胆碱能神经元的相关性 在灵长类动物的排卵和无排卵期间。 资助 NIH HD16631， HD18185 出版物 Pau CY、Pau KYF、Spies HG。 假定的雌激素 雄性和雌性恒河猴中的受体和 mRNA 表达。 分子细胞内分泌学 146：59-68，1998。