Zhen Wei, Li Peng, He Bin, Guo Juyuan, Zhang Yong-Lian
Shanghai Key Laboratory for Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Biol Reprod. 2009 Apr;80(4):696-706. doi: 10.1095/biolreprod.108.071589. Epub 2008 Dec 17.
We describe a novel epididymis-specific cDNA named Glb1l4, which was isolated from rat epididymis by differential display of mRNAs. Glb1l4 cDNA contains 2607 nucleotides and encodes a 637-amino acid protein with 50% similarity to mouse beta-galactosidase. The gene is located on chromosome 8q13, spanning 21 exons. Northern blot analysis reveals that Glb1l4 is specifically expressed in the caput region of epididymis and upregulated by androgen. A specific polyclonal antiserum against the N-terminal peptide of GLB1L4 has been produced. Western blot analysis and immunohistochemistry assay reveal that GLB1L4 is specifically expressed in the principal cells of the caput epididymis. Interestingly, its expression peaks at Postnatal Day 45 in mRNA level and at Postnatal Day 60 in protein level while the epididymis column cells undergo differentiation. Moreover, within this very period this secretory protein is confined inside the cell with a change of subcellular distribution pattern, which implies its important roles in the cell differentiation process. Only after the epididymal epithelium differentiation is completed and the spermatozoa enter the epididymal lumen is the GLB1L4 secreted into the luminal fluid and bound on the sperm head. Our results suggest that GLB1L4 may play various roles in principal cell differentiation and sperm maturation.
我们描述了一种新的附睾特异性cDNA,命名为Glb1l4,它是通过mRNA差异显示从大鼠附睾中分离出来的。Glb1l4 cDNA包含2607个核苷酸,编码一个637个氨基酸的蛋白质,与小鼠β-半乳糖苷酶有50%的相似性。该基因位于8号染色体q13上,跨越21个外显子。Northern印迹分析表明,Glb1l4在附睾头部区域特异性表达,并受雄激素上调。已制备了针对GLB1L4 N端肽的特异性多克隆抗血清。蛋白质印迹分析和免疫组织化学检测表明,GLB1L4在附睾头部的主细胞中特异性表达。有趣的是,其在出生后第45天mRNA水平达到峰值,在出生后第60天蛋白质水平达到峰值,而附睾柱状细胞在此期间进行分化。此外,在这个时期内,这种分泌蛋白被限制在细胞内,亚细胞分布模式发生变化,这意味着它在细胞分化过程中起重要作用。只有在附睾上皮分化完成且精子进入附睾管腔后,GLB1L4才分泌到管腔液中并结合在精子头部。我们的结果表明,GLB1L4可能在主细胞分化和精子成熟中发挥多种作用。