Role of translation by mitochondrial‐type ribosomes during sperm capacitation: An analysis based on a proteomic approach
Role of translation by mitochondrial‐type ribosomes during sperm capacitation: An analysis based on a proteomic approach
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DOI：
10.1002/pmic.200800353
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发表时间：
2009-03
期刊：
影响因子：
3.4
通讯作者：
Chun Zhao;Xuejiang Guo;Zhongqing Shi;F. Wang;Xiao-yan Huang;R. Huo;Hui Zhu;Xin-ru Wang;Jia-yin Liu;Zuo-min Zhou;J. Sha
中科院分区：
文献类型：
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作者：
Chun Zhao;Xuejiang Guo;Zhongqing Shi;F. Wang;Xiao-yan Huang;R. Huo;Hui Zhu;Xin-ru Wang;Jia-yin Liu;Zuo-min Zhou;J. Sha
哺乳动物精子含有复杂的 mRNA 群，其中一些已被证明可以通过线粒体型核糖体使用 D-氯霉素 (CP)（一种线粒体翻译的特异性抑制剂）从头翻译。然而，人们对这些 mRNA 在成熟精子中的功能知之甚少。在本研究中，采用差异蛋白质组学方法来研究使用抑制剂 CP 由线粒体型核糖体翻译的精子蛋白谱，并鉴定出 44 种蛋白质在经 CP 处理的精子中表达低于获能精子（比率 ≥ 1.5，p<0.05）。蛋白质印迹和实时 PCR 的结果表明四种蛋白质是由线粒体型核糖体翻译的。生物信息学分析表明，44 个蛋白质中有 26 个参与了与精卵相互作用事件相关的一些关键过程。此外，Mups的繁殖功能从未被研究过，因此被选择进行进一步研究。我们的结果表明，Mups 蛋白定位于预获能精子的顶体和鞭毛，并且也在获能精子的赤道段表达。使用中和抗体消耗 Mups 以剂量依赖性方式显着抑制获能，随后抑制顶体反应和精卵融合。总之，获能过程中的线粒体翻译可以储存有利于精子与卵子相互作用的蛋白质。
Mammalian spermatozoa contain a complex population of mRNAs, some of which have been demonstrated to be translated de novo by mitochondrial‐type ribosomes using D‐chloramphenicol (CP), a specific inhibitor of mitochondrial translation. However, little is known about the functions of these mRNAs in mature sperm. In the present study, differential proteomic approaches were applied to study sperm protein profiles translated by mitochondrial‐type ribosomes using the inhibitor CP and 44 proteins were identified with lower expression in CP‐treated sperm in comparison to capacitated sperm (ratio ≥ 1.5, p<0.05). Results of Western blot and real‐time PCR suggest that four proteins were translated by mitochondrial‐type ribosomes. Bioinformatics analysis indicated that 26 of 44 proteins were involved in some critical processes correlated to sperm–egg interaction event. In addition, Mups, whose functions in reproduction have never been studied, were chosen for further study. Our results showed that Mups proteins were localized to the acrosome and flagellum of precapacitated sperm, and were also expressed in the equatorial segment of capacitated sperm. The depletion of Mups using neutralizing antibodies significantly inhibited capacitation in a dose‐dependent manner, subsequently inhibited acrosome reaction and sperm–egg fusion. In summary, mitochondrial translation during capacitation can store proteins beneficial for sperm–egg interaction.