LH AND FSH SECRETION BY INDIVIDUAL PITUITARY CELLS

各个垂体细胞分泌 LH 和 FSH

• 批准号: 3234969
• 负责人: JIMMY D NEILL
• 金额: $ 12.03万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-04-01 至 1989-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3234969
• 关键词: antiserum; electron microscopy; estrus; follicle stimulating hormone; gonadotropin releasing factor; hemolysis; histochemistry /cytochemistry; hormone regulation /control mechanism; immunochemistry; immunoperoxidase; luteinizing hormone; pituitary gland; pituitary gonadal axis; plaque assay; secretion

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?

目的是研究功能和形态学的程度 促性腺激素的异质性是FSH和LH的模式的基础 分泌及其对性腺类固醇，GNRH和抑制素的调节 在大鼠的发情周期中。 要解决的主要问题 是垂体中是否有促性腺激素的亚群 腺体以及这些子群是否在功能能力上改变 在发情循环中，以说明前溶前LH和FSH 激增，以及LH和FSH的差分分泌。 我开发了一种检测个人激素分泌的方法 混合文化中的细胞为解决这个问题提供了希望。 它 是反向溶血斑块测定法，是基于补体介导的 蛋白A耦合RBC的裂解与垂体孵育 细胞和激素抗血清。 RBC裂解的结果区域（“斑块”） 垂体细胞周围很容易在显微镜下识别。 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 要进行比较。 具体目的是：1）单个大鼠促性腺激素分泌的LH分泌 使用反向溶血斑块测定，电子显微镜评估， 和免疫细胞化学来回答以下问题：什么 促性腺激素的一部分在各种生理状态下分泌LH？ b。 做相同的促性腺激素同时显示抑制作用和刺激作用 LH分泌的雌激素？ 2）单个老鼠的FSH分泌 促性腺激素将使用斑块测定法评估电子显微镜检查 和免疫细胞化学来回答以下问题：什么 AP细胞的比例是各种生理学期间的FSH分泌剂 和实验状态？ b。在优先的FSH分泌过程中，是FSH 由仅FSH促性腺激素或FSH+LH Gonadotropes分泌？ c。哪个 Gonadotrope细胞类型可作用于优先抑制FSH的作用 分泌？