FETAL MEMBRANE MATRIX METALLOPROTEINASES IN MONKEYS SPONTANE & ACTH INDUCE LABO

排卵前促性腺激素释放激素（GnRH）的释放 女性和恒河猴的促黄体生成激素（LH）依赖于 卵巢雌二醇-17的分泌增强后， （E）。 这种E诱导 促性腺激素释放激素/促黄体生成素激增伴有下丘脑和脑干的变化 细胞被鉴定为神经肽Y（NPY）-，去甲肾上腺素 (NE)-、乙酰胆碱（Ach）-和γ-氨基丁酸 （GABA）神经元。 我们的研究集中在E 影响与这些基因相关的转录-翻译事件， 肽和递质系统以及这些中枢神经事件 随着年龄的增长而变化，特别是在围绝经期生活阶段， 分泌下降。 E受体的两种亚型（ER？和？）有 被举报。 为了研究ER亚型的组织分布， 我们克隆了恒河猴的两个cDNA探针（126 bp和291 bp）并进行了序列测定 猴子急诊室。 这些cDNA片段类似，但不相同， 人类急诊室？ 序列的 两个人都有问题-？ 和ER-？，我们 定量ER亚型在几个外周和中枢神经 通过逆转录聚合酶链测定雄性和雌性组织 反应（RT-PCR）。 两个急诊室？ 而ER-？ mRNA存在于 男性和女性生殖器官。 只有呃-？ 与会者表示 肝脏、额叶皮质、尾状核、蓝斑和 小脑 在某些大脑区域，即，内壳核 囊，海马和室旁下丘脑，ER-？ 是 在雌性中表达，但在雄性中不表达。 在雌性猕猴中 10-15岁酪氨酸羟化酶（TH， 用于控制NE合成的酶）在特定肾上腺素能神经元中 群体（A1，A2，A6）通过E处理上调。 然而，在这方面， 在围绝经期猴（> 22岁）中E治疗后， TH mRNA的表达水平在A6区被抑制， 脑干 我们继续研究这个时代的相关性， 脑干神经元TH mRNA反应的E相关下降。 我们也 克隆了一个356 bp的猴胆碱乙酰转移酶（CHAT， Ach合成的控制）cDNA片段，该cDNA片段对应于人 CHAT N-1型序列332至684。 5=和3=引物序列 分别为TAAGATGGCAGCAAAAACTCCC和ACCGATGACCAGCTGAGGTTT。 据我们所知，恒河猴CHAT的部分DNA序列 基因以前没有记录。 MCHAT RNA探针， 从与CHAT mRNA杂交的克隆中产生的几种特异性 成年雌性猕猴脑中的下丘脑和脑干区域。 在各种生理过程中mRNA表达的定量 条件将有助于建立胆碱能神经元的相关性 在灵长类动物排卵和不排卵期间。 资助NIH HD 16631， 出版物Pau CY，Pau KYF，Spies HG. 推定雌激素 受体和mRNA的表达。 Mol Cell Endocrinol 146：59-68，1998.

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.