CRES GENE IN MALE REPRODUCTION

我们研究的总体目标是确定小说的作用 克雷斯蛋白在男性生殖系统中的表达 功能 克雷斯蛋白表现出一种非常有限的模式， 表达，并在mRNA水平上受到睾丸激素的高度调节。 因素 在检测的25个组织中，雄性特异性克雷斯mRNA/蛋白表达量为1.5%，而雄性特异性CRES mRNA/蛋白表达量为1.5%。 仅局限于头的近端区域 附睾，睾丸的圆形/伸长的精子细胞，以及 脑垂体中的特定细胞 戏剧性的外观和 睾丸和附睾中克雷斯蛋白的消失表明 这种蛋白质可能在一个非常离散的 睾丸和附睾功能的阶段。此外， 将克雷斯蛋白转化为半胱氨酸蛋白酶抑制剂的半胱氨酸蛋白酶抑制剂家族， 所有这些都是分泌性结合蛋白，这表明它也可能 具有蛋白质结合活性。事实上，我们的初步研究表明， 克雷斯蛋白特异性结合近端头的头部 附睾精子在一个非常短暂的相互作用。所以我们的 一种假说是克雷斯蛋白可能是一种调节结合蛋白 精子发生和精子成熟所必需的。具体地说， 所述克雷斯蛋白可以结合精子相关蛋白，可能是 蛋白酶然后，这种结合可能导致结合的“激活”。 伴侣，随后可能是关键的生精和精子 成熟的事件为了验证我们的假设，我们将：1）扩展我们的克雷斯 通过检查细胞和细胞内的蛋白定位研究 克雷斯蛋白在睾丸、附睾、垂体和 在光镜和电镜水平上观察精子; 2）分离精子， 大鼠克雷斯cDNA，并制备大鼠克雷斯抗体用于表征 大鼠克雷斯基因和蛋白; 3）开始克雷斯蛋白表征研究 通过检查克雷斯蛋白的翻译后修饰， 半胱氨酸蛋白酶抑制剂活性;和4）检查克雷斯蛋白与 小鼠和大鼠精子，并启动研究，以确定克雷斯蛋白 结合伴侣这些研究将提供有价值的信息， 一种新的附睾蛋白，可能对精子发育很重要， 成熟

The overall objective of our studies is to determine the role of the novel CRES (cystatin-related epididymal-specific) protein in male reproductive function. The CRES protein exhibits a remarkably restricted pattern of expression and is highly regulated at the mRNA level by testicular factors. Of the 25 tissues examined, the male-specific CRES mRNA/protein has been localized to only the very proximal region of the caput epididymidis, the round/elongating spermatids of the testis, and to specific cells in the pituitary. The dramatic appearance and disappearance of the CRES protein in the testis and epididymis suggests that the protein may perform a specialized role during a very discrete phase of testicular and epididymal function. Furthermore, the homology of the CRES protein to the cystatin family of cysteine protease inhibitors, all of which are secretory binding proteins, suggests that it also may have protein binding activity. Indeed our preliminary studies suggest that CRES protein binds specifically to the head of the proximal caput epididymal spermatozoa in a very transient interaction. Therefore, our hypothesis is that the CRES protein may be a regulatory binding protein necessary for spermatogenic and sperm maturational events. Specifically, the CRES protein may bind to a sperm-associated protein, perhaps a protease. This binding may then result in the "activation" of the binding partner which subsequently may be critical for spermatogenic and sperm maturational events. To test our hypothesis we will: 1) extend our CRES protein localization studies by examining the cellular and intracellular localization of the CRES protein in the testis, epididymis, pituitary, and spermatozoa at the light and electron microscopic levels; 2) Isolate the rat CRES cDNA and prepare a rat CRES antibody for characterization of the rat CRES gene and protein; 3) begin CRES protein characterization studies by examining post-translational modifications of the CRES protein and cystatin activity; and 4) examine the interaction of CRES protein with mouse and rat sperm and initiate studies to identify the CRES protein binding partner. These studies will provide valuable information on a novel epididymal protein which may be important for sperm development and maturation.