Localization and expression of DNAJB8 in male germ cells
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摘要:
目的：探究DNAJB8［DnaJ heat shock protein family（HSP40） member B8］在小鼠精子发生过程中的表达模式。方法：分别采用实时荧光定量PCR和蛋白免疫印迹实验检测小鼠各脏器、不同发育时期睾丸组织以及不同生精细胞DNAJB8的mRNA转录和蛋白表达量；免疫荧光检测DNAJB8在生精细胞中的表达定位。结果：DNAJB8基因在睾丸组织中特异表达，从小鼠出生后21 d（P21）即圆形精子阶段开始DNAJB8 mRNA保持较高的表达，而DNAJB8蛋白在精子变形晚期出生后35 d（P35）才开始表达。DNAJB8蛋白主要定位在晚期精子细胞及精子鞭毛中段。结论：DNAJB8可能参与小鼠精子变形晚期的发育过程。
Abstract:
Objective：This study aims to investigate the expression pattern of heat shock protein DNAJB8［DnaJ heat shock protein family（HSP40）member B8］ in spermatogenesis. Methods：Quantitative real-time PCR and Western blot were used to determine the transcript levels and protein abundance of DNAJB8 in various mouse tissues，testes at different developmental stages and different germ cell populations. The subcellular localization of DNAJB8 was further assessed by immunofluorescence analysis. Results：DNAJB8 was specifically expressed in the mouse testis. The transcripts of DNAJB8 mRNA increased significantly at postnatal day 21（P21），when round spermatids appeared. However，the protein of DNAJB8 was detected until in P35 mouse testis，where elongating spermatids were highly enriched at this stage. Immunostaining on frozen adult mouse testis and spermatozoa showed that DNAJB8 was observed in late stages of spermatids，and its signal was also present in middle-piece flagellum of spermatozoa. Conclusion：DNAJB8 may function in the elongation of round spermatids during late stages of spermiogenesis.
