The identification of stallions and or ejaculates that will provide commercially acceptable quality post-thaw before cryopreservation is of great interest, avoiding wasting time and resources freezing ejaculates that will not achieve sufficient quality to be marketed. Our hypothesis was that after bioinformatic analysis, the study of the stallion sperm proteome can provide discriminant variables able to predict the post-thaw quality of the ejaculate. At least three ejaculates from 10 different stallions were frozen following a split sample design. Half of the ejaculate was analyzed as a fresh aliquot and the other half was frozen and then analyzed as a frozen-thawed aliquot. Computer-assisted sperm analysis and flow cytometry were used to analyze sperm quality. Detailed proteomic analysis was performed on fresh and frozen and thawed aliquots, and bioinformatic analysis was used to identify discriminant variables in fresh samples able to predict the outcome of cryopreservation. Those with a fold change > 3, a P = 8.2e-04, and a q = 0.074 (equivalent to False discovery rate (FDR)) were selected, and the following proteins were identified in fresh samples as discriminant variables of good motility post-thaw: F6YTG8, K9K273, A0A3Q2I7V9, F7CE45, F6YU15, and F6SKR3. Other discriminant variables were also identified as predictors of good mitochondrial membrane potential and viability post-thaw. We concluded that proteomic approaches are a powerful tool to improve current sperm biotechnologies.

中文翻译：

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种马精子蛋白质组的差异解释了种马间精子质量在解冻后的变异性†

对冷冻保存前解冻后能提供商业上可接受质量的种马和/或精液的鉴定是非常重要的，以避免浪费时间和资源冷冻精液而不能达到足够的质量以上市销售。我们的假设是，经过生物信息学分析，种马精子蛋白质组的研究可以提供能够预测解冻后射精质量的判别变量。在拆分样本设计之后，来自 10 匹不同种马的至少三份精液被冷冻。一半的精液作为新鲜的等分试样进行分析，另一半被冷冻，然后作为冷冻解冻的等分试样进行分析。计算机辅助精子分析和流式细胞仪用于分析精子质量。对新鲜、冷冻和解冻的等分试样进行了详细的蛋白质组学分析，生物信息学分析用于识别能够预测冷冻保存结果的新鲜样品中的判别变量。那些有倍数变化的人 > 3、选择了a P = 8.2e-04，aq = 0.074（相当于错误发现率（FDR）），在新鲜样品中鉴定出以下蛋白质作为解冻后良好运动性的判别变量：F6YTG8、K9K273、 A0A3Q2I7V9、F7CE45、F6YU15 和 F6SKR3。其他判别变量也被确定为解冻后良好线粒体膜电位和活力的预测因子。我们得出结论，蛋白质组学方法是改进当前精子生物技术的有力工具。选择074（相当于错误发现率（FDR）），并在新鲜样品中鉴定出以下蛋白质作为解冻后良好运动性的判别变量：F6YTG8、K9K273、A0A3Q2I7V9、F7CE45、F6YU15和F6SKR3。其他判别变量也被确定为解冻后良好线粒体膜电位和活力的预测因子。我们得出结论，蛋白质组学方法是改进当前精子生物技术的有力工具。选择074（相当于错误发现率（FDR）），并在新鲜样品中鉴定出以下蛋白质作为解冻后良好运动性的判别变量：F6YTG8、K9K273、A0A3Q2I7V9、F7CE45、F6YU15和F6SKR3。其他判别变量也被确定为解冻后良好线粒体膜电位和活力的预测因子。我们得出结论，蛋白质组学方法是改进当前精子生物技术的有力工具。