Cornwall G A, Hsia N
Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, Lubbock 79430, USA.
Endocrinology. 1997 Oct;138(10):4262-72. doi: 10.1210/endo.138.10.5468.
The maturation of spermatozoa in the epididymis is a complex process that requires the active involvement of the epididymal epithelium. The primary focus toward elucidating the role of the epididymis in the maturation process has been the study of epididymal secretory proteins and their interaction with spermatozoa. To date there is a paucity of information regarding epididymal epithelial cell surface proteins, which may also play important roles in epididymal function. Through a subtractive hybridization approach to identify genes specifically expressed in the caput epididymidis, the mouse homologue of a member of the ADAM (a disintegrin and metalloprotease) family of proteins was identified. This rapidly growing gene family encodes cell surface proteins that possess putative adhesion and protease domains. Northern blot analyses demonstrated that the mouse ADAM gene, termed ADAM7, is expressed in the caput region of the epididymis and in the anterior pituitary gonadotropes with no detectable expression in the twenty-six other tissues examined. Furthermore, in situ hybridization experiments revealed that the ADAM7 messenger RNA (mRNA) exhibited an apical localization within the proximal caput epididymal epithelium that may correlate with an unusual sparsely granulated endoplasmic reticulum uniquely present in the proximal region of the epididymidis and to which no known function has been ascribed. Hormonal, surgical, and genetic strategies demonstrated that ADAM7 gene expression requires, in a region-dependent manner, androgens as well as testicular factors for expression. Interestingly, the apical localization of ADAM7 mRNA is dependent upon an intact testis, because in situ hybridization analyses of the proximal caput epididymidis from a testosterone maintained castrate mouse did not show the apical localization of ADAM7 mRNA. Finally, chromosomal mapping demonstrated that the ADAM7 gene maps to the central region of mouse Chromosome 14, approximately 4-5 cM distal from the fertilin beta locus, which encodes another reproductive-specific ADAM protein.
精子在附睾中的成熟是一个复杂的过程,需要附睾上皮的积极参与。阐明附睾在成熟过程中作用的主要研究方向是附睾分泌蛋白及其与精子相互作用的研究。迄今为止,关于附睾上皮细胞表面蛋白的信息匮乏,而这些蛋白可能在附睾功能中也发挥着重要作用。通过消减杂交方法来鉴定在附睾头中特异性表达的基因,我们鉴定出了一种ADAM(去整合素和金属蛋白酶)蛋白家族成员的小鼠同源物。这个快速增长的基因家族编码具有假定粘附和蛋白酶结构域的细胞表面蛋白。Northern印迹分析表明,小鼠ADAM基因(称为ADAM7)在附睾头区域和垂体前叶促性腺细胞中表达,而在所检测的其他26种组织中未检测到表达。此外,原位杂交实验显示,ADAM7信使核糖核酸(mRNA)在附睾近端头上皮细胞的顶端定位,这可能与附睾近端区域独特存在的异常稀疏颗粒内质网相关,且该内质网尚无已知功能。激素、手术和遗传学策略表明,ADAM7基因表达以区域依赖的方式需要雄激素以及睾丸因子来表达。有趣的是,ADAM7 mRNA的顶端定位依赖于完整的睾丸,因为对睾酮维持的去势小鼠附睾近端头进行原位杂交分析未显示ADAM7 mRNA的顶端定位。最后,染色体定位表明,ADAM7基因定位于小鼠14号染色体的中央区域,距离编码另一种生殖特异性ADAM蛋白的受精素β基因座约4 - 5厘摩。