LH AND FSH SECRETION BY INDIVIDUAL PITUITARY CELLS

其目的是调查功能和形态的程度 促性腺激素的异质性是卵泡刺激素和促黄体生成素模式的基础 性腺激素、促性腺激素释放激素和抑制素的分泌及其调节 在老鼠的发情周期中。要解决的主要问题 脑下垂体中是否存在促性腺激素亚群 腺体，以及这些亚群的功能能力是否发生变化 在发情周期中，以解释排卵前的黄体生成素和卵泡刺激素 促黄体生成素和促卵泡刺激素的不同分泌。 我开发了一种检测个体荷尔蒙分泌的方法 混合培养中的细胞，这为解决这个问题提供了希望。它 是基于补体介导的反向溶血空斑试验 脑垂体单层孵育的A蛋白偶联红细胞的裂解 细胞和激素抗血清。由此产生的红细胞裂解带(“斑块”) 在显微镜下可以很容易地辨认出脑垂体细胞周围。 随后的电子显微镜(或EM免疫细胞化学) 功能鉴定的细胞允许形态和功能 需要进行比较。 具体目标是：1)个别大鼠促性腺激素分泌黄体生成素 用反向溶血空斑分析，电子显微镜， 和免疫细胞化学来回答以下问题：a.什么 促性腺激素部分在各种生理状态下分泌黄体生成素？B. 同样的促性腺激素是否同时表现出抑制和刺激作用？ 雌激素对黄体生成素分泌的影响？2)大鼠个体卵泡刺激素的分泌 促性腺激素将使用菌斑分析、电子显微镜进行评估。 和免疫细胞化学来回答以下问题：a.什么 AP细胞在不同生理状态下分泌FSH的比例 那么实验州呢？B.在FSH优先分泌期间，是FSH 是由仅有FSH的促性腺激素分泌的，还是由FSH+LH促性腺激素分泌的？C.哪一项 促性腺激素细胞类型抑制素是否作用于优先抑制FSH 分泌物？

The aim is to investigate the extent to which functional and morphological heterogeneity of gonadotropes underlies the patterns of FSH and LH secretion and their regulation by gonadal steroids, GnRH, and inhibin during the estrous cycle of the rat. The major questions to be addressed are whether there are sub-populations of gonadotropes in the pituitary gland, and whether these sub-populations change in functional capacity during the estrous cycle so as to account for the preovoluatory LH and FSH surges, and for the differential secretion of LH and FSH. I have developed a method for detecting hormone secretion by individual cells in mixed cultures which offers hope for resolution of this issue. It is the reverse hemolytic plaque assay and is based on complement mediated lysis of protein A coupled RBC incubated in a monolayer with pituitary cells and hormone antiserum. The resulting zones of RBC lysis ("plaques") surrounding the pituitary cell are easily identified microscopically. Subsequent electron microscopy (or EM immunocytochemistry) on such functionally identified cells allows morphological and functional comparisons to be made. The specific aims are: 1) LH secretion by individual rat gonadotropes will be assessed using the reverse hemolytic plaque assay, electron microscopy, and immunocytochemistry to answer the following questions: a. What fraction of the gonadotropes secrete LH in various physiologic states? b. Do the same gonadotropes show both the inhibitory and stimulatory effects of estrogen on LH secretion? 2) FSH secretion by individual rat gonadotropes will be assessed using the plaque assay, electronmicroscopy and immunocytochemistry to answer the following questions: a. What proportion of the AP cells are FSH secretors during various physiological and experimental states? b. During preferential FSH secretion, is FSH secreted by FSH-only gonadotropes or by FSH+LH gonadotropes? c. Which gonadotrope cell type does inhibin act on to preferentially inhibit FSH secretion?