PDF(Pubmed)
Abstract：
At present, there are few reports about the proteomics changes provoked by butylated hydroxytoluene (BHT) supplementation on cryopreserved semen in mammals. Thus, we aimed to evaluate the effects of different concentrations of BHT on goat sperm and to investigate the proteomics changes of adding BHT to cryopreserved goat (Capra hircus) sperm. Firstly, semen samples were collected from four goats, and frozen in the basic extenders containing different concentrations of BHT (0.5 mM, 1.0 mM, 2.0 mM) and a control without BHT, respectively. After thawing, the protective effects of dose-dependent replenished BHT to the freezing medium on post-thaw sperm motility, integrities of plasma membrane and acrosome, reactive oxygen species levels were confirmed, with 0.5 mM BHT being the best (B group) as compared to the control (without BHT, C group). Afterwards, TMT-based quantitative proteomic technique was performed to profile proteome of the goat sperm between C group and B group. Parallel reaction monitoring was used to confirm reliability of the data. Overall, 2,476 proteins were identified and quantified via this approach. Comparing the C and B groups directly (C vs. B), there were 17 differentially abundant proteins (DAPs) po-tentially associated with sperm characteristics and functions were identified, wherein three were upregulated and 14 were downregulated, respectively. GO annotation analysis demonstrated the potential involvement of the identified DAPs in metabolic process, multi-organism process, reproduction, reproductive process, and cellular process. KEGG enrichment analysis further indicated their potential roles in renin-angiotensin system and glutathione metabolism pathways. Together, this novel study clearly shows that BHT can effectively improve quality parameters and fertility potential of post-thawed goat sperm at the optimal concentration, and its cryoprotection may be realized through regulation of sperm metabolism and antioxidative capability from the perspective of sperm proteomic modification.
摘要：
目前,关于补充丁基化羟基甲苯（BHT）对哺乳动物冷冻保存的精液引起的蛋白质组学变化的报道很少。因此，我们旨在评估不同浓度的BHT对山羊精子的影响，并研究在冷冻保存的山羊（Caprahircus）精子中添加BHT的蛋白质组学变化。首先,从四只山羊中收集精液样本，并冷冻在含有不同浓度BHT（0.5mM，1.0mM,2.0mM）和不含BHT的对照，分别。解冻后,剂量依赖性补充BHT对冷冻后精子运动的保护作用，质膜和顶体的完整性，确认了活性氧的水平，与对照组（无BHT，C组）。之后,采用基于TMT的定量蛋白质组学技术对C组和B组山羊精子进行蛋白质组分析。平行反应监测用于确认数据的可靠性。总的来说,通过这种方法鉴定和定量2,476种蛋白质。直接比较C组和B组（Cvs.B),确定了17种与精子特征和功能相关的差异丰富蛋白（DAP），其中3个上调，14个下调，分别。GO注释分析表明，已识别的DAP可能参与代谢过程，多有机体过程，繁殖，生殖过程，细胞过程。KEGG富集分析进一步表明它们在肾素-血管紧张素系统和谷胱甘肽代谢途径中的潜在作用。一起,这项新的研究清楚地表明，在最佳浓度下，BHT可以有效地改善解冻后山羊精子的质量参数和育性潜力，从精子蛋白质组修饰的角度来看，其冷冻保护作用可能是通过调节精子代谢和抗氧化能力来实现的。
