Molecular biologic analysis of gonadotropin genes in patients with ovulation disorders caused by pituitary dysfunction

垂体功能障碍所致排卵障碍患者促性腺激素基因的分子生物学分析

基本信息

批准号：
08457440
负责人：
SUGANUMA Nobuhiko
金额：
$ 5.12万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

项目摘要

Female reproductive function is regulated by hypothalamic-pituitary-ovarian system. Pulsatile released gonadotropin-releasing hormone (GnRH) from hypothalamus stimulates the secretion of pituitary gonadotropins (FSH and LH) that not only induce ovarian follicle growth and ovulation but also produce ovarian steroid hormones (estrogen and progesterone) . The syntheses of these hormones are regulated each other through the feed-back mechanism. Abnormalities in this system causes ovulation disorders. In the patients with pituitary dysfunction, abnormalities in the following parts are speculated : 1) Response to GnRH (GnRH receptor), 2) Signal transduction after binding of GnRH to GnRH receptor, 3) Regulatory elements for gonadotropin gene expression, and 4) Gonadotropin genes. Thus, we tried to detect the causes of ovulation disorders by pituitary dysfunction in the present research project.Six female patients with ovulation disorders by pituitary dysfunction were prepared at Nagoya University Hospital. Lymphocytes were collected from the patients, and DNAs were extracted by guanidium method. Genes of GnRH receptor, LHbeta-subunit, and 300-bp upstream of gonadotropin alpha-subunit were amplified by PCR technic using these DNA as templates. The amplified DNA was cloned to plasmid through TA cloning and sequenced. As results, no abnormalities were found in the above genes in all the patients.Additionally, new findings in anomalous LH and relationship between structure and function of gonadotropin subnits were gotten. Moreover, gonadotropin effects to some gene expression in oocyte and granulosa cells and clinical significance of GnRH agonist were also studied.

女性生殖功能受下丘脑 - 垂体 - 伏雄叶的调节。从下丘脑释放促性腺激素释放激素（GNRH）的脉冲运动刺激垂体促性腺激素（FSH和LH）的分泌，不仅会诱导卵巢卵泡生长和排卵，还产生卵巢固醇激素（雌激素和叶酸）。这些激素的合成通过喂养机制相互调节。该系统中的异常会导致排卵障碍。在垂体功能障碍的患者中，猜测以下部分异常：1）对GNRH的反应（GNRH受体），2）2）GNRH与GNRH受体结合后的信号转导，3）3）调节性元素的促性腺素基因表达和4）Gonadotropin基因。因此，我们试图在本研究项目中检测出垂体功能障碍的排卵障碍原因。在纳戈亚大学医院制备了垂体功能障碍的六种女性排卵疾病的女性患者。从患者中收集淋巴细胞，并通过鸟翼法提取DNA。 GNRH受体，LHBETA-SUBUNIT和300 bp上游的促性腺激素α-亚基的基因使用这些DNA作为模板进行了放大。通过TA克隆将放大的DNA克隆至质粒并测序。结果，在所有患者中未发现上述基因中的异常。此外，还研究了促性腺激素对卵母细胞和颗粒细胞中某些基因表达的作用以及GnRH激动剂的临床意义。