TISSUE SPECIFIC EXPRESSION AND FUNCTION OF ME-RABP

精子在附睾中成熟，获得受精能力， 体内，并在射精前储存。 维生素A是重要的， 附睾功能，因为它需要维持上皮细胞 结构. 视黄酸受体α的靶向突变 （RAR α）导致附睾上皮鳞状化生 导致不孕症（1）。类维生素A是疏水不稳定的 化合物在水环境中运输， 载体蛋白 我们已经发现，限制性上皮细胞 附睾区域（远端头）分泌雄激素依赖性 附睾腔内的维甲酸结合蛋白（E-RABP） 管道。 我们已经克隆，测序并定位在2号染色体上， 鼠E-RABP基因。 我们已经证明，5 '侧翼区的5 kb 连接在CAT报告基因的前面，靶向一个令人惊讶的高水平的 头端的基因表达水平。 我们假设mE-RABP 对于类维生素A在体内的稳态和运输至关重要。 附睾，其区域特异性表达由 附睾转录因子 我们的目标是建立转基因 小鼠模型1）识别和表征最小序列 赋予mE-RABP基因表达的区域特异性，和2） 破坏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 雄激素受体的区域特异性表达的mE- 附睾内的RABP基因。 2)确定监管要素 及其相关的参与附睾的结合蛋白-以及 mE-RABP基因的区域特异性表达。 3)确定 使用mE-RABP的缺失和取代变体的mE-RABP的功能 转基因小鼠模型中的RABP基因。建成后将 为将来的研究提供了有价值的工具，旨在探讨附睾 功能 mE-RABP启动子可用于在体内破坏其他细胞因子。 附睾基因允许人们分析这些基因的功能作用， 附睾中的基因 mE-RABP基因敲除小鼠可用于重新构建 在异位部位（尾部或输精管）表达mE-RABP， 确定区域特异性基因的功能重要性， 在附睾中的表达。 这一计划是我们长期 目的是了解基因表达的机制， 附睾 这项提议是我们长期目标的一部分， 附睾提供最佳环境的机制 男性配子，并有助于生产一个肥沃的射精。 最终目的是为理性者提供基础知识 治疗某些形式的男性不育症。

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.