EXPRESSION OF NEUROPEPTIDE Y MRNA IN ESTROGEN TREATED RHESUS MACAQUES

雌激素处理的恒河猴中神经肽 Y mRNA 的表达

• 批准号: 6277379
• 负责人: HAROLD G SPIES
• 金额: $ 7.42万
• 依托单位: OREGON REGIONAL PRIMATE RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6277379
• 关键词: Mammalia; Primates; biological products; clinical research; endocrine gland /system; male; nervous system; nutrition; nutrition related tag; reproductive system; women's health

Neuropeptide Y (NPY) modulates cardiovascular, feeding and reproductive functions. Peripheral neurohumoral inputs from these systems are integrated and transformed into efferent signals in the brainstem. However, detailed mapping of NPY-expressing cells in the brainstem has not been established in primates. We have previously shown that central infusion of NPY stimulates a massive release of hypothalamic gonadotropin-releasing hormone (GnRH), suggesting that increased NPY gene expression, synthesis and release may be responsible in part for the initiation and/or maintenance of the estradiol-17 (E2)-induced preovulatory GnRH surge in primate species. Bi-directional neurohumoral communication between the brainstem and hypothalamus has been well established, and E2 receptor binding has been demonstrated in the monkey brainstem. Thus, we hypothesize that exogenous E2 treatment, which is known to induce a preovulatory-like surge of luteinizing hormone (LH), will increase the expression of NPY mRNA in the brainstem. To test this hypothesis, we utilized the in situ hybridization method to identify and quantitate NPY mRNA in ovariectomized rhesus macaques without or with E2 treatment. Individual brains were fixed with paraformaldehyde before (n=5) or at 20 h (n=3) and 30 h (n=5) after the E2 treatment time intervals that span the initiation and peak plasma LH surge. In the brainstem, hybridized images of NPY mRNA were found in the nucleus of the solitary tract, the dorsal motor nucleus of the vagus nerve, the nucleus of the spinal tract of trigeminal nerve, the lateral reticular nucleus, the reticular formation, the locus coeruleus, the nucleus of raphe and the lateral tegmentum surrounding the periaqueductal grey. To compare the expression of NPY mRNA between E2 treated and untreated animals, hybridized signals were transferred onto Cronex X-ray films and digitized with NIH image software. Signals/cell were quantified by a computer-assisted image analysis program. Examination of the locus coeruleus shows that the optical densities of NPY mRNA expression after 20 h (113 q 21) and 30 h (128 q 17) were significantly (p< 0.01) higher than those in E2-untreated animals (23 q 11). Comparison of NPY mRNA expression in other brain areas is in progress. These data suggest that estrogen may enhance gene expression in brainstem NPY neurons. The up-regulated mRNA expression in the locus coeruleus may be an important component in initiating and/or sustaining the preovulatory GnRH release in the hypothalamus.

神经肽Y（NPY）调节心血管、摄食和 生殖功能。 外周神经体液输入来自这些 系统被集成并转换为传出信号， 脑干 然而，在胰腺癌中表达NPY的细胞的详细定位， 脑干尚未在灵长类动物中建立。 我们先前已经 表明中枢神经肽Y的注入刺激了大量的 下丘脑促性腺激素释放激素（GnRH），这表明， 可能是神经肽Y基因表达、合成和释放增加 部分负责发起和/或维持 雌二醇-17（E2）诱导灵长类动物排卵前GnRH激增。 脑干和小脑之间的双向神经体液通讯 下丘脑已经很好地建立，E2受体结合已经 在猴子脑干中得到证实。 因此，我们假设， 外源性E2治疗，这是已知的诱导排卵前样 促黄体生成素（LH）的激增，会增加NPY的表达 脑干中的mRNA。 为了验证这一假设，我们使用了 用原位杂交方法检测了脑组织中NPY mRNA的表达 切除卵巢的恒河猴，没有或有E2治疗。 将个体脑用多聚甲醛固定，然后（n=5）或 E2处理时间间隔后20 h（n=3）和30 h（n=5）， 跨越血浆LH峰的起始和峰值。 在脑干， 结果显示，NPY mRNA的杂交图像出现在细胞核内。 孤束，迷走神经背侧运动核， 三叉神经脊束外侧网状核 核，网状结构，蓝斑， 中缝和中脑导水管周围灰质周围的外侧被盖。 比较E2处理组与未处理组之间NPY mRNA的表达 动物，杂交信号转移到Cronex X射线胶片上 并用NIH图像软件进行数字化。 定量信号/细胞 通过计算机辅助图像分析程序。 审查 蓝斑显示NPY mRNA的光密度 20 h（113 q21）和30 h（128 q17）后的表达， 显著（p< 0.01）高于E2未处理的动物（23 问题11）。 比较其他脑区的NPY mRNA表达， 中求进工作总 这些数据表明，雌激素可能会增强基因表达， 在脑干神经肽Y神经元中的表达。 上调的mRNA表达 在蓝斑可能是一个重要的组成部分， 和/或维持下丘脑中排卵前GnRH的释放。