HORMONE ACTION ON GABA/GLUTAMATE BALANCE OF GNRH CELLS

描述 父母补助金将使用形态测量学，免疫组织化学，定量 突触学和原位杂交研究雌激素的作用 对灵长类动物GnRH神经元的影响。 本申请是 扩展相同的目的，使用预嵌入双标记 光和EM免疫细胞化学促性腺激素释放激素和六种递质之一， 是抑制性的（GABA、儿茶酚胺、β-内啡肽）或兴奋性的 （甘丙肽、P物质、神经肽Y）对GnRH释放的影响。 此外，本发明还提供了一种方法， 植入后GABA和谷氨酸的免疫细胞化学将用于 定量它们各自对GnRH神经元的输入。 显微照片将是 定量接触GnRH阳性的染色和未染色终扣， 阴性细胞，使用体视学方法在5 每组实验动物。 这些小组将由灵长类动物组成， 月经周期中期的内源性或模拟雌激素激增，以及手术 或自然绝经的动物，有些被雌激素取代。 类似 实验将在卵巢切除的大鼠上进行，用雌二醇治疗 或控制。 一般来说，灵长类动物的治疗和组织增生 将在美国实验室进行，并在两个实验室进行大鼠实验。 的 免疫细胞化学和分析将在外来部位进行。

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.