TISSUE SPECIFIC EXPRESSION AND FUNCTION OF ME-RABP

在附睾精子成熟时，获得受精的能力 在体内，并在射精前储存。维生素A对人体健康很重要 附睾功能，因为它需要维持上皮细胞 结构。视黄酸受体α基因的靶向突变 (RARpha)导致附睾上皮鳞状化生 并导致不孕症(1)。类维甲酸是疏水不稳定的 化合物在水环境中运输，结合到特定的 载体蛋白。我们发现，一种限制性的上皮细胞 附睾区(头端远端)分泌雄激素依赖性激素 附睾腔内视黄酸结合蛋白的表达 风管。我们已经克隆、测序并定位在2号染色体上 小鼠E-RABP基因。我们已经证明了5‘侧翼区的5kb 在猫记者面前连接的基因靶标高得惊人 头部的基因表达水平。我们的假设是Me-RABP 对于维甲酸在体内的动态平衡和运输是必不可少的 它的区域特异性表达是由 附睾转录因子。我们的目标是建立转基因 小鼠模型1)识别和刻画最小序列 Me-RABP基因表达的区域特异性和2)对 干扰Me-RABP基因表达以确定Me-RABP的功能 蛋白。本研究的具体目的是：1)确定角色 雄激素受体在Me-Me的区域特异性表达 附睾腺内有RABP基因。2)确定监管要素 以及与附睾体相关的结合蛋白--以及 Me-RABP基因的区域特异性表达。3)确定 利用Me-RABP的缺失和替换变体实现Me-RABP的功能 RABP基因在转基因小鼠模型中的表达。该项目的完成将 为未来的附睾体研究提供有价值的工具 功能。Me-RABP启动子可用于在体内干扰其他 附睾基因使人们能够分析这些基因的功能作用 附睾体中的基因。Me-RABP基因敲除小鼠可用于重新检测 在异位部位(尾部或输精管)表达我-RABP，允许 确定区域特异性基因的功能重要性 在附睾部的表达。这项建议是我们长期计划的一部分 目的了解基因在细胞中表达的机制。 附睾症。这项建议是我们长期目标的一部分，以了解 附睾为人类提供最佳环境的机制 雄性配子，并有助于产生可生育的射精。 最终目的是为理性的人提供基本的知识 治疗某些形式的男性不育。

In the epididymis spermatozoa mature, gain the ability to fertilize in vivo and are stored prior to ejaculation. Vitamin A is important in epididymal function since it is required to maintain epithelial structures. Targeted mutation of the retinoic acid receptor alpha (RARalpha) results in squamous metaplasia of the epididymal epithelium with resulting infertility (1). The retinoids being hydrophobic labile compounds are transported in a aqueous environments bound to specific carrier proteins. We have found that the epithelium of a restricted region of the epididymis (distal caput) secretes an androgen dependent retinoic acid binding protein (E-RABP) in the lumen of the epididymal duct. We have cloned, sequenced and localized on chromosome 2 the murine E-RABP gene. We have shown that 5 kb of the 5'flanking region ligated in front of the CAT reporter gene targets a surprisingly high level of gene expression in the caput. Our hypothesis is that mE-RABP is essential for the homeostasis and trafficking of retinoids within the epididymis and that its region-specific expression is determined by epididymal transcription factors. Our goals are to establish transgenic mouse models 1) to identify and characterize the minimal sequences conferring region specificity to mE-RABP gene expression and 2) to disrupt mE-RABP gene expression to determine the function of the mE-RABP protein. The specific aims of this study are: 1) To determine the role of the androgen receptor in the region-specific expression of the mE- RABP gene within the epididymis. 2) To identify the regulatory elements and their associated binding proteins involved in the epididymis-and region-specific expression of the mE-RABP gene. 3) To determine the function of mE-RABP using deletion and substitution variants of the mE- RABP gene in transgenic mouse models. Completion of the project will provide valuable tools for future studies designed to probe epididymal function. The mE-RABP promoter can be used to disrupt in vivo other epididymal genes allowing one to analyze the functional role of these genes in the epididymis. The mE-RABP knock-out mice can be used to re- express mE-RABP in an ectopic site (cauda or vas deferens), allowing one to determine the functional importance of region-specific gene expression in the epididymis. This proposal is part of our long-term goal to understand the mechanisms by which the gene expression in the epididymis. This proposal is part of our long-term goal to understand the mechanisms by which the epididymis provides the optimal milieu for male gametes and contributes to the production of a fertile ejaculate. The ultimate goal is to provide the basic knowledge for the rational treatment of some forms of male infertility.