Teperek Marta, Miyamoto Kei, Simeone Angela, Feret Renata, Deery Michael J, Gurdon John B, Jullien Jerome
Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, UK.
Cambridge Centre for Proteomics, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
Int J Mol Sci. 2014 Sep 19;15(9):16719-40. doi: 10.3390/ijms150916719.
Spermatozoa are more efficient at supporting normal embryonic development than spermatids, their immature, immediate precursors. This suggests that the sperm acquires the ability to support embryonic development during spermiogenesis (spermatid to sperm maturation). Here, using Xenopus laevis as a model organism, we performed 2-D Fluorescence Difference Gel Electrophoresis (2D-DIGE) and mass spectrometry analysis of differentially expressed proteins between sperm and spermatids in order to identify factors that could be responsible for the efficiency of the sperm to support embryonic development. Furthermore, benefiting from the availability of egg extracts in Xenopus, we also tested whether the chromatin of sperm could attract different egg factors compared to the chromatin of spermatids. Our analysis identified: (1) several proteins which were present exclusively in sperm; but not in spermatid nuclei and (2) numerous egg proteins binding to the sperm (but not to the spermatid chromatin) after incubation in egg extracts. Amongst these factors we identified many chromatin-associated proteins and transcriptional repressors. Presence of transcriptional repressors binding specifically to sperm chromatin could suggest its preparation for the early embryonic cell cycles, during which no transcription is observed and suggests that sperm chromatin has a unique protein composition, which facilitates the recruitment of egg chromatin remodelling factors. It is therefore likely that the acquisition of these sperm-specific factors during spermiogenesis makes the sperm chromatin suitable to interact with the maternal factors and, as a consequence, to support efficient embryonic development.
与精子不成熟的直接前体细胞精子细胞相比,精子在支持正常胚胎发育方面效率更高。这表明精子在精子发生过程中(精子细胞到精子成熟)获得了支持胚胎发育的能力。在这里,我们以非洲爪蟾作为模式生物,对精子和精子细胞之间差异表达的蛋白质进行了二维荧光差异凝胶电泳(2D-DIGE)和质谱分析,以确定可能导致精子支持胚胎发育效率的因素。此外,得益于非洲爪蟾卵提取物的可用性,我们还测试了与精子细胞的染色质相比,精子的染色质是否能吸引不同的卵因子。我们的分析确定了:(1)几种仅存在于精子中而不存在于精子细胞核中的蛋白质;以及(2)在卵提取物中孵育后,大量与精子(而非精子细胞染色质)结合的卵蛋白。在这些因素中,我们鉴定出许多与染色质相关的蛋白质和转录抑制因子。转录抑制因子特异性结合精子染色质的现象可能表明其为早期胚胎细胞周期做准备,在此期间未观察到转录现象,这也表明精子染色质具有独特的蛋白质组成,有利于募集卵染色质重塑因子。因此,很可能在精子发生过程中获得这些精子特异性因子使精子染色质适合与母体因子相互作用,从而支持高效的胚胎发育。