HORMONE ACTION ON GABA/GLUTAMATE BALANCE OF GNRH CELLS

描述 父母的资助将使用形态测量，免疫组织化学，定量 突触学和原位杂交技术研究雌激素的作用 关于灵长类GnRH神经元的输入。本申请是一种 相同目的的延伸，使用预嵌入双标记 GnRH和六种递质之一的光和EM免疫细胞化学 抑制性(GABA、儿茶酚胺、β-内啡肽)或兴奋性 (甘丙肽、P物质、神经肽Y)释放GnRH。此外, 包埋后GABA和谷氨酸的免疫细胞化学将用于 量化它们各自对GnRH神经元的输入。显微照片将会是 对与GnRH阳性接触的染色和未染色的部位进行定量 阴性细胞，使用体视学方法进行量化 每组实验动物。这些团体将由#年的灵长类组成。 月经中期内源性或模拟雌激素激增，以及外科手术 或者是自然更年期的动物，其中一些被雌激素取代。类似 实验将在去卵巢的大鼠身上进行，用雌二醇治疗 或者控制。总的来说，灵长类动物的治疗和组织获取 将在美国实验室进行，并在两个实验室进行老鼠实验。这个 将在异地进行免疫细胞化学和分析。

DESCRIPTION The parent grant will use morphometry, immunohistochemistry, quantitative synaptology and in situ hybridization to investigate the effect of estrogen on the input to GnRH neurons of primates. The present application is an extension of the same aims, with the use of pre-embedding double labeled light and EM immunocytochemistry for GnRH and one of six transmitters that are either inhibitory (GABA, catecholamines, beta-endorphin) or excitatory (galanin, substance P, neuropeptide Y) to GnRH release. In addition, postembedding immunocytochemistry for GABA and glutamate will be used to quantitate their respective inputs to GnRH neurons. The micrographs will be quantified for stained and unstained boutons contacting GnRH positive and negative cells, using a stereological method for quantification in 5 experimental animals per group. The groups will be comprised of primates in endogenous or simulated estrogen surge of mid menstrual cycle, and surgical or naturally menopausal animals, some replaced with estrogen. Similar experiments will be performed on ovariectomized rats, treated with estradiol or control. In general, the treatments and tissue procurements in primates will be performed at the US lab, and rat experiments at both labs. The immunocytochemistry and analysis will be performed at the foreign site.