绵羊精子冻融前后差异蛋白质组学研究

摘要

[目的]检测和分析绵羊精子冻融前后的差异蛋白质.[方法]用二维凝胶电泳分离精子蛋白,凝胶用考马斯亮蓝染色,用PDQues t8.0软件检测分析差异蛋白质斑点并经液相色谱串联离子阱质谱鉴定.[结果]鲜冻精凝胶图谱分析后得到14个差异蛋白斑点,在国际生物信息学中心数据库搜索到相对应的5种蛋白质:推定的热休克蛋白70.1、转录因子AP-2 α蛋白、烯醇酶、烯醇酶1和血清白蛋白前体.在解冻精子中,推定的热休克蛋白70.1、烯醇酶和烯醇酶1含量减少,转录因子AP-2α蛋白含量增加,血清白蛋白前体来源于精浆.[结论]绵羊精子经冷冻、解冻后,蛋白质存在差异,对差异蛋白质的研究有利于探索绵羊精子在冷冻过程中的生理状态,为进一步研究绵羊精子蛋白质组学及揭示精子冻伤机理提供依据.%[Objective] To investigate the differential proteins between fresh and frozen-thawed sheep sperm. [Method] The current research presents the protein changes using two-dimensional gel electrophoresis (2-DE) after stained with commassie blue, differential proteins were detected by PDQuest 8.0 software and subjected to ion trap mass spectrometer equipped with a Surveyor HPLC system, and differential spots of protein were identified. [Result] A search against protein sequences in the National Center for Biotechnology Information databases indicated that the differential fourteen proteins correspond to five proteins including putative heat shock protein 70.1, transcription factor AP-2-alpha, enolase, enolasel and serum albumin precursor that protein content of putative heat shock protein 70.1, enolase, enolasel were decreased and protein content of transcription factor AP-2-alpha was increased in frozen-thawed spermatozoa, serum albumin precursor was seminal plasma protein. [Conclusion] It was concluded there was a significant difference at protein level in sheep sperm during frozen and frozen-thawed. Differential proteins analysis may be useful to clarify the physiology state of sheep sperm during cryopreservation for further studies on sheep sperm proteomic and revealing the cryodamage mechanism of sperm cryopreservation.