LH AND FSH SECRETION BY INDIVIDUAL PITUITARY CELLS

目的是调查功能和形态 促性腺激素的异质性是FSH和LH模式的基础 分泌及其由性腺类固醇激素、GnRH和Escherichin调节 在大鼠的发情周期中。 需要解决的主要问题 垂体中是否有促性腺激素亚群 腺体，以及这些亚群的功能能力是否发生变化 以解释排卵前LH和FSH 激增，以及LH和FSH的差异分泌。 我发明了一种检测个体激素分泌的方法 混合培养中的细胞，这为解决这一问题提供了希望。 它 是反向溶血空斑试验，基于补体介导的 与垂体单层孵育的蛋白A偶联红细胞的溶解 细胞和激素抗血清。 所产生的RBC溶解区（“斑块”） 在显微镜下很容易识别。 随后的电子显微镜（或EM免疫细胞化学）对这种 功能鉴定的细胞允许形态和功能 要进行比较。 具体的目的是：1）单个大鼠促性腺激素分泌LH， 使用反向溶血空斑试验，电子显微镜， 和免疫细胞化学来回答以下问题：a.什么 促性腺激素在不同生理状态下分泌LH的比例？ B. 相同的促性腺激素是否同时表现出抑制和刺激作用 对LH分泌的影响 2)单个大鼠的FSH分泌 促性腺激素将使用空斑试验、电子显微镜 和免疫细胞化学来回答以下问题：a.什么 在各种生理过程中，AP细胞的比例是FSH分泌者。 实验性国家？ B.在FSH优先分泌期间，FSH 是由FSH促性腺激素分泌还是由FSH+LH促性腺激素分泌？ C.这 促性腺激素细胞类型是否影响FSH的优先抑制作用 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?