家蚕精巢中小热激蛋白在不同发育时期的蛋白质组学分析

Proteome Analysis of Small Heat Shock Proteins in Bombyx mori Testis at Different Developmental Stages

【目的】研究家蚕不同发育时期精巢组织中小热激蛋白(sHSP)变化,为阐明其在家蚕精子发生中的重要作用提供理论参考。【方法】利用双向电泳技术建立家蚕5龄第5天、化蛹第1天和化蛾第1天3个时期的精巢电泳图谱,并用Imagemaster2D 6.0软件分析不同时期图谱中的差异蛋白,再用基质辅助激光解析离子质谱(MALDI-TOF-MS)对这些蛋白进行鉴定。【结果】鉴定出家蚕精巢中5个小热激蛋白,分别为sHSP23.7、sHSP20.8、sHSP 20.4、sHSP19.9和sHSP19.8,它们在化蛹第1天和化蛾第1天的蛋白合成量比5龄第5天显著降低。【结论】在家蚕精巢中,5龄第5天是形成有核精子的重要时期,有大量的微管蛋白(α微管蛋白、β微管蛋白),它是中心粒和纺锤体的重要组成成分,在减数分裂中有重要作用,这些小热激蛋白可能在有核精子形成时与微管蛋白相互作用,在促进鞭毛轴丝形成和鞭毛运动方面发挥重要作用。

【Objective】 The objective of this study is to investigate the developmental changes in the expression of major small heat shock proteins（sHSP） in Bombyx mori testis and to provide a theoretical basis for the molecular mechanism of sHSPs in spermatogenesis.【Method】 The electronphoretogram of testis from day 5 of fifth instar,day 1 of pupa and day 1 moth were established by two-dimensional polyacrylamide gel electrophoresis.Moreover,Imagemaster2D 6.0 was performed to analyze the different expression patterns of these sHSPs and these proteins were identi？ed by matrix-assistedlaser desorption/ionization mass spectrometry（MALDI-TOF-MS）.【Result】 Five sHSPs（sHSP23.7,sHSP20.8,sHSP20.4,sHSP19.9 and sHSP19.8） were successfully identified on two-dimensional gels.And it was found that the level of protein biosyntesis of these five sHSPs in the day 1 of pupa and day 1 moth reduced so obviously compared with the day 5 of fifth instar.【Conclusion】 Since the day 5 of fifth instar is the important period to form the eupyrene sperm,there are a large number of tubulins（α-tubulin and βtubulin）,which are not only the major component of centriole and spindle,but also play an important role in meiosis.It is concluded that the sHSPs may interact with the tubulin when the eupyrene sperm is forming and play an important role in the speeding of the axoneme formation and flagellar movement.