HOX GENES IN THE ADULT MOUSE EPIDIDYMIS AND VAS DEFERENS

精子在附睾中的成熟需要上皮基因产物，以特定区域的模式表达。这项建议的长期目标是发现成人附睾节段功能维持的潜在机制。我们假设HOX基因是这一过程的主要调节者。众所周知，HOX基因在胚胎的节段性模式中起着至关重要的作用，但成人的功能尚不清楚。我们将使用RT-PCR和Northern分析来研究选定的HOX基因在成年小鼠附睾体中的表达水平。具体而言，Hoxa-11蛋白的区域差异和细胞定位将分别用Western印迹分析和免疫组织化学方法进行研究。同样的方法将被用于检测Meis 1，已知的HOX基因的DNA结合辅助因子，以及细胞黏附分子L1-CAM，Hoxa-11蛋白的潜在下游目标。Hoxa-11、辅因子和下游靶在成年大鼠附睾中的相互作用将用凝胶迁移率漂移试验进行分析。这些研究探索了一种新的、特定的途径，与维持附睾部的功能分段有关。这一信息将为附睾功能提供新的见解，对于理解男性生育和不育非常重要。

Sperm maturation in the epididymis requires epithelial gene products, expressed in a region specific pattern. The long-term objective of this proposal is to discover the mechanism underlying the maintenance of segmental function in the adult epididymis. We hypothesize that hox genes are master regulators of this process. Hox genes are known to be crucial in segmental patterning of the embryo but adult function is unknown. Expression levels of selected hox genes will be studied in the adult mouse epididymis using RT-PCR and Northern analysis. Regional differences and cellular localization of hoxa-11 protein, specifically, will be studied with Western blot analysis and immunohistochemistry, respectively. The same methods will be used to examine for Meis 1, a known DNA-binding cofactor for hox genes, and for the cell adhesion molecule L1-CAM, a potential downstream target for hoxa-11 protein. Interaction of hoxa-11, cofactor and downstream target in the adult rat epididymis will be analyzed by Electrophoretic Mobility Shift Assay. These studies investigate a novel, specific pathway relevant in the maintenance of functional segmentation of the epididymis. This information will provide new insights into epididymal function, important to the understanding of male fertility and infertility.