Grignard Elise, Morin Joelle, Vernet Patrick, Drevet Joel R
Laboratoire Epididyme & Maturation des Gamètes, Université Blaise Pascal, UMR CNRS 6547-GEEM, 24 avenue des Landais, 63177 Aubière cedex, France.
Theriogenology. 2005 Sep 1;64(4):1016-33. doi: 10.1016/j.theriogenology.2005.01.008. Epub 2005 Mar 5.
We report here on the cloning of cDNAs coding bovine and equine orthologs of mouse epididymis-restricted and sperm-bound glutathione peroxidase 5 (GPX5), a selenium-independent member of the multigenic GPX family in mammals. The complete sequence of bovine GPX5 as well as a partial sequence of the equine GPX5 were characterized, conceptually translated and aligned with other known mammalian GPX5 proteins. Using Northern blotting assays, we show that the level of expression of GPX5 is high in bovine but low in equine and that in both species the regionalization of GPX5 expression in epididymis is not totally identical to what was reported for rodent mouse GPX5. An antibody was produced against GPX5 and used in Western blot assays as well as in immunohistochemistry assays on bovine epididymis sections. It shows that the protein is essentially present in the cytoplasmic compartment of the caput segment 2 epithelium of the bovine epididymis. Unlike in the mouse model, bovine GPX5 seems to be poorly secreted and does not seem to be present on cauda epididymal spermatozoa.
我们在此报告了编码小鼠附睾限制性和精子结合型谷胱甘肽过氧化物酶5(GPX5)的牛和马直系同源物的cDNA的克隆情况,GPX5是哺乳动物多基因GPX家族中一种不依赖硒的成员。对牛GPX5的完整序列以及马GPX5的部分序列进行了表征、概念性翻译,并与其他已知的哺乳动物GPX5蛋白进行了比对。通过Northern印迹分析,我们发现GPX5在牛中的表达水平较高,而在马中较低,并且在这两个物种中,GPX5在附睾中的表达区域化与啮齿动物小鼠GPX5的报道情况并不完全相同。制备了针对GPX5的抗体,并将其用于蛋白质印迹分析以及牛附睾切片的免疫组织化学分析。结果表明,该蛋白主要存在于牛附睾头段2上皮细胞的细胞质区室中。与小鼠模型不同,牛GPX5似乎分泌较少,并且在附睾尾精子上似乎不存在。