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。本研究的长期目标是发现成人附睾节段功能维持的机制。我们假设hox基因是这一过程的主要调节者。已知Hox基因在胚胎的片段模式中起关键作用，但成人的功能尚不清楚。选择hox基因的表达水平将在成年小鼠附睾使用RT-PCR和Northern分析进行研究。具体来说，将分别用Western blot分析和免疫组织化学方法研究hoxa-11蛋白的区域差异和细胞定位。同样的方法将用于检测Meis 1（已知的hox基因的dna结合辅助因子）和细胞粘附分子L1-CAM （hoxa-11蛋白的潜在下游靶点）。用电泳迁移位移法分析成年大鼠附睾中hoxa-11、辅助因子和下游靶点的相互作用。这些研究探讨了一种新的、特定的途径，与维持附睾的功能分割有关。这一信息将为了解附睾功能提供新的见解，对了解男性生育能力和不育症具有重要意义。

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.