Analysis of Signal Transduction of Growth and Differentiation of Avian Granulosa cells

禽颗粒细胞生长和分化的信号转导分析

基本信息

批准号：
05806035
负责人：
MORI Makoto
金额：
$ 1.22万
依托单位：
Shizuoka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05806035/
关键词：
Quail Aves Follicle Ovulation Steroid Granulosa Ovary 卵巣顆粒膜細胞プロゲステロンホルモン鳥類

项目摘要

Granulosa cells of follicles in avian species, such as chicken and Japanese quail, . proliferate during follicular growth, and differentiate to secrete progesterone in response to luteinizing hormone (LH), when the follicle becomes the largest and reaches to the time of ovulation. In order to study the mechanism and signal transduction pathway of growth and differentiation, quail granulosa cells were cultured with various gonadotrophins or growth factors, such as epidermal growth factor (EGF).The culture medium was a mixture of McCoy's 5a and Ham's F12 (1 : 1) supplemented with glutamine, HEPES,Penicilline and Streptomycin. The concentration of fetal calf serum was 10% during the initial 6 hr of the culture, and then it was reduced to 1%. In order to evaluate the LH response of the cells after the culture, the cells were incubated with or without LH for 3 hr in Krebs-Ringer solution supplemented with bovine serum albumin, and progesterone production was measured in the medium during this 3 hr of incubation by the specific radio immunoassay.When the granulosa cells were cultured with follicle stimulating hormone (FSH) for 72 hr, the response to LH was increased while the cell numbers were not significantly increased. When the granulosa cells were cultured with EGF for 72 hr, the cell numbers increased dramatically, but the LH response of the cells, which was evaluated by the production of progesterone, was diminished. These results suggested that the growth and differentiation of the granulosa cells during follicular maturation was controlled by both FSH and EGF.

鸟类（例如鸡和日本鹌鹑）的卵泡颗粒细胞。在卵泡生长过程中增殖，并在卵泡达到最大并达到排卵时，响应黄体生成素（LH）而分化并分泌黄体酮。为了研究生长和分化的机制和信号转导途径，将鹌鹑颗粒细胞与各种促性腺激素或生长因子如表皮生长因子（EGF）一起培养。培养基为McCoy's 5a和Ham's F12的混合物（1 ：1）补充谷氨酰胺、HEPES、青霉素和链霉素。培养最初6小时内胎牛血清浓度为10%，随后降至1%。为了评估培养后细胞的 LH 反应，将细胞在有或没有 LH 的情况下在补充有牛血清白蛋白的 Krebs-Ringer 溶液中孵育 3 小时，并在这 3 小时的孵育期间测量培养基中的孕酮产量当颗粒细胞与卵泡刺激素（FSH）一起培养72小时时，对LH的反应增加，但细胞数量没有明显增加。当颗粒细胞与 EGF 一起培养 72 小时时，细胞数量急剧增加，但细胞的 LH 反应（通过黄体酮的产生来评估）却减弱。这些结果表明，卵泡成熟过程中颗粒细胞的生长和分化受到 FSH 和 EGF 的控制。