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，其以区域特异性模式表达。本研究的长期目标是探索成人附睾节段性功能维持的机制。我们假设hox基因是这一过程的主要调节者。已知Hox基因在胚胎的节段模式中至关重要，但成人功能尚不清楚。将使用RT-PCR和北方分析在成年小鼠附睾中研究所选hox基因的表达水平。hoxa-11蛋白的区域差异和细胞定位，具体来说，将分别用Western印迹分析和免疫组化进行研究。同样的方法将用于检测Meis 1（一种已知的hox基因的DNA结合辅因子）和细胞粘附分子L1-CAM（一种hoxa-11蛋白的潜在下游靶点）。将通过电泳迁移率变动试验分析hoxa-11、辅因子和下游靶标在成年大鼠附睾中的相互作用。这些研究探讨了一种新的，特定的途径相关的维持功能分割的附睾。这些信息将为附睾功能提供新的见解，对理解男性生育力和不育症很重要。

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.