STRUCTURE AND NEGATIVE REGULATION OF THE CLUSTERIN GENE

丛生蛋白基因，也称为硫酸化糖蛋白（SGP-2）基因， 在睾丸中以组织特异性方式表达和调节，雄性 附属性器官和许多其他组织 的表达 雄激素对大鼠clusterin基因表达有负调控作用 前列腺和cAMP在睾丸细胞。 虽然cAMP的能力， 刺激基因表达已在许多系统中进行了研究，但所知甚少 关于cAMP在几种Leydig肿瘤细胞系中的负面作用， 作为一个独特的模型系统来研究可能的机制 参与cAMP对基因表达的细胞特异性抑制。 在 在本项目中，我们建议研究clusterin基因的结构， 这种表达的负面荷尔蒙控制。 我们的第一个具体目标 是为了表征大鼠丛生蛋白基因的结构，特别是， 该基因的5-侧翼区域。 第二个具体目标是确定 细胞所需的DNA元件和蛋白质因子- 睾丸细胞和卵巢上皮细胞中簇蛋白基因的特异性基础表达， 男性附属性器官 我们将检查是否有一个消极的 雄激素反应元件负责抑制该基因 前列腺腹侧的雄激素 我们的第三个具体目标是调查 参与的机制（转录活性和mRNA稳定性） 负cAMP反应元件和蛋白质因子， 负责cAMP在Leydig肿瘤细胞系中的抑制作用 以及Leydig和Sertoli细胞的原代培养物中。 最后我们将 研究促性腺激素，通过cAMP和雄激素起作用，是否也 在体内对聚集蛋白mRNA和蛋白水平产生抑制作用， 大鼠睾丸间质细胞。 据信，青蒿素能发挥保护作用， 对雄性生殖道细胞和生殖细胞起作用。 的 信息，将获得从这个项目上的激素 该基因的调控将有助于我们理解 clusterin可能参与雄性生殖功能。

The clusterin gene, also known as the sulfated glycoprotein (SGP-2) gene, is expressed and regulated in a tissue-specific manner in the testis, male accessory sex organs, and many other tissues. The expression of the clusterin gene was shown to be negatively regulated by androgens in rat prostate and by cAMP in testicular cells. Although the ability of cAMP to stimulate gene expression has been studied in many systems; little is known about the negative effect of cAMP in several Leydig tumor cell lines can be used as a unique model system to investigate the possible mechanisms involved in the cell-specific inhibition of gene expression by cAMP. In this project, we propose to study the structure of the clusterin gene and the negative hormonal control of this expression. Our first specific aim is to characterize the structure of the rat clusterin gene, in particular, the 5-flanking region of the gene. The second specific aim is to determine the DNA elements and protein factors that are required for the cell- specific basal expression of the clusterin gene in testicular cells and in the male accessory sex organs. We will examine whether a negative androgen-responsive element is responsible for the suppression of this gene by androgens in ventral prostate. Our third specific aim is to investigate the mechanisms (transcriptional activity and mRNA stability) involved in the negative cAMP-responsive elements and the protein factors that are responsible for the inhibitory effect of cAMP in Leydig tumor cell lines and in primary cultures of Leydig and Sertoli cells. Finally, we will investigate whether gonadotropins, acting through cAMP, and androgens also exert inhibitory effects in vivo on clusterin mRNA and protein levels in Leydig cells of rat testis. Clusterin is believed to exert a protective function on cells of the male reproductive tract and on germ cells. The information that will be obtained from this project on the hormonal regulation of this gene will facilitate our understanding of the role that clusterin may play in the male reproductive function.