Neesen Jürgen, Hartwich Tobias, Brandhorst Gunnar, Aumüller Gerhard, Gläser Birgitta, Burfeind Peter, Mendoza-Lujambio Irene
Institute of Human Genetics, University of Göttingen, Heinrich-Düker-Weg 12, Göttingen 37073, Germany.
Biochem Biophys Res Commun. 2002 Oct 4;297(4):737-48. doi: 10.1016/s0006-291x(02)02265-9.
To identify new genes that could be involved in the differentiation and function of male germ cells, we have screened a murine testis cDNA library and isolated a clone that was named Tep22. The gene encoding Tep22 consists of three exons and is localized in the telomeric region of mouse chromosome 12. Expression analyses with RNA from different adult tissues revealed that Tep22 is predominantly expressed in spermatocytes and spermatids of the murine testis. Four Tep22 transcript sizes ranging from 647 to 1122 nucleotides were detected in testes of 15-day-old mice due to variable 5' UTRs, while the open reading frame of Tep22 has a length of 567bp in all transcript forms. Specific antibodies against Tep22 detected an approximately 22kDa band in testicular protein extracts, which was first observed in 18-day-old mice, indicating that Tep22 is translationally repressed for several days. Indirect immunofluorescence and immunoelectron microscopy experiments demonstrate that Tep22 is localized in the acrosomal region of early elongating spermatids, while the surrounding cytoplasm is barely labeled. During further germ cell development, the intensity of the staining in the acrosomal region decreases and is no longer detectable in late stages of elongating spermatids, whereas the amount of the Tep22 protein increases in the cytoplasm. Finally, Tep22 is incorporated into the midpiece of spermatids and is also present in the mitochondrial sheath of mature spermatozoa. Taken together, our results suggest that Tep22 is involved in the biogenesis of the acrosome as well as in the function of the midpiece of murine spermatozoa.
为了鉴定可能参与雄性生殖细胞分化和功能的新基因,我们筛选了小鼠睾丸cDNA文库并分离出一个名为Tep22的克隆。编码Tep22的基因由三个外显子组成,定位于小鼠染色体12的端粒区域。对来自不同成年组织的RNA进行的表达分析表明,Tep22主要在小鼠睾丸的精母细胞和精子细胞中表达。由于5'非翻译区可变,在15日龄小鼠的睾丸中检测到四种大小从647到1122个核苷酸的Tep22转录本,而Tep22的开放阅读框在所有转录本形式中的长度均为567bp。针对Tep22的特异性抗体在睾丸蛋白提取物中检测到一条约22kDa的条带,该条带最早在18日龄小鼠中观察到,表明Tep22在翻译水平上被抑制了几天。间接免疫荧光和免疫电子显微镜实验表明,Tep22定位于早期伸长精子细胞的顶体区域,而周围的细胞质几乎没有标记。在生殖细胞进一步发育过程中,顶体区域的染色强度降低,在伸长精子细胞后期不再可检测到,而Tep22蛋白在细胞质中的量增加。最后,Tep22被整合到精子细胞的中段,并且也存在于成熟精子的线粒体鞘中。综上所述,我们的结果表明Tep22参与了小鼠精子顶体的生物发生以及中段的功能。