Gene expression of gonadotropin and its producing pituitary cells

促性腺激素及其产生垂体细胞的基因表达

• 批准号: 06454019
• 负责人: KATO Yukio
• 金额: $ 4.35万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454019/
• 关键词: reproduction; pituitary; hormone; gonadotropin; gene expression; transcription factor; DNA binding protein; 生殖刺激ホルモン; 転写調節; ゴナドトロピン; ブタ

In this project that focused the molecular biological study on the gene expression of gonadotropin and its producing pituitary cells, following results were obtained.1.The transcription factors, cJun and cFos, were cloned from the anterior pituitary.2.The involvement in the GnRH-dependent modulation of mRNA levels of cJun and cFos were clarified by quantitative RT-PCR.3.According to the identify of pituitary cJun and cFos, yeast two-hybrid system that could search the interaction factor (s) for cJun were performed. From the screening of 40,000 cDNA clones, a new pituitary transcription factor, ATF4, was obtained, revealing that pituitary cJun forms heterodimer with ATF4 and participates with modulation of pituitary gene expressions.4.A pituitary specific transcription factor, Pit-1, that is essential for tissue specifc expression of GH gene as well as differentiation of pituitary somatoroph, mammotroph thyrotroph, was cloned from porcine pituitary and its nucleotide sequence was determined.5.Yeast one-hybrid system to identify factors that recognize regulatory sequence of hormone genes were constructed and 25 clones were obtained as candidates. Re-transformation to yeast cells showed that they are false positives. The cloning is now in progress.

本课题主要针对促性腺激素及其产生垂体细胞的基因表达进行分子生物学研究，取得了以下成果：1.从垂体前叶克隆了转录因子cJun和cFos。2.GnRH的参与通过定量RT-PCR阐明了cJun和cFos mRNA水平的依赖性调节。3.根据垂体cJun和cFos的鉴定，酵母双杂交执行了可以搜索 cJun 相互作用因子的系统。通过4万个cDNA克隆的筛选，获得了一个新的垂体转录因子ATF4，揭示垂体cJun与ATF4形成异二聚体，参与垂体基因表达的调节。 4.垂体特异性转录因子Pit-1，即从猪中克隆出对于 GH 基因的组织特异性表达以及垂体生长细胞、促乳腺激素细胞的分化至关重要5.构建了识别激素基因调控序列的酵母单杂交系统，获得了25个候选克隆。重新转化酵母细胞表明它们是假阳性。克隆工作正在进行中。

项目成果

Kato, Y.: "Gene structure of eukaryote" Gakkai-shupan Centar, Molecular Biology of Hormone. Vol 1. 18 (1996)

Kato, Y.：“真核生物的基因结构”Gakkai-shupan Centar，激素分子生物学。

玄浩一郎: "性成熟と配偶子形成に働くホルモン" 化学と生物. 33. 784-792 (1995)

Koichiro Gen：“影响性成熟和配子发生的激素”化学与生物学 33. 784-792 (1995)。

加藤幸雄: "組換え体性腺刺激ホルモンの生産と応用" 獣医畜産新報. 49. 39-43 (1996)

加藤幸雄：“重组促性腺激素的生产和应用”《兽医与畜牧杂志》49. 39-43 (1996)。

加藤幸雄: "「ホルモンの分子生物学(日本比較内分泌学会編)」シリーズ 第3巻 生殖とホルモン 第2章ゴナドトロピン" 学会出版センター(印刷中),

加藤幸雄：《激素的分子生物学（日本比较内分泌学会编）》丛书第3卷《生殖与激素》第2章促性腺激素》学会出版中心（印刷中），

加藤幸雄: "ゴナドトロピン遺伝子の発現調節機構" Hormone Frontier in Gynecology. 1. 19-29 (1994)

加藤幸雄：“促性腺激素基因表达调控机制”妇科激素前沿。1. 19-29 (1994)。