The Scripps Research Institute, La Jolla, CA, USA.
Federal University of Viçosa, Viçosa, MG, Brazil.
Sci Rep. 2020 Sep 4;10(1):14661. doi: 10.1038/s41598-020-71015-9.
The present study investigated the seminal plasma proteome of Holstein bulls with low (LF; n = 6) and high (HF; n = 8) sperm freezability. The percentage of viable frozen-thawed sperm (%ViableSperm) determined by flow cytometry varied from -2.2 in LF to + 7.8 in HF bulls, as compared to the average %ViableSperm (54.7%) measured in an 860-sire population. Seminal proteins were analyzed by label free mass spectrometry, with the support of statistical and bioinformatics analyses. This approach identified 1,445 proteins, associated with protein folding, cell-cell adhesion, NADH dehydrogenase activity, ATP-binding, proteasome complex, among other processes. There were 338 seminal proteins differentially expressed (p < 0.05) in LF and HF bulls. Based on multivariate analysis, BSP5 and seminal ribonuclease defined the HF phenotype, while spermadhesin-1, gelsolin, tubulins, glyceraldehyde-3-phosphate dehydrogenase, calmodulin, ATP synthase, sperm equatorial segment protein 1, peroxiredoxin-5, secretoglobin family 1D and glucose-6-phosphate isomerase characterized the LF phenotype. Regression models indicated that %ViableSperm of bulls was related to seminal plasma peroxiredoxin-5, spermadhesin-1 and the spermadhesin-1 × BSP5 interaction (R = 0.84 and 0.79; p < 0.05). This report is the largest dataset of bovine seminal plasma proteins. Specific proteins of the non-cellular microenvironment of semen are potential markers of sperm cryotolerance.
本研究调查了精子冷冻能力低（LF；n=6）和高（HF；n=8）的荷斯坦公牛的精浆蛋白质组。通过流式细胞术测定的活冷冻解冻精子的百分比（%ViableSperm）在 LF 公牛中为-2.2，在 HF 公牛中为+7.8，而在 860 头公牛的群体中平均 %ViableSperm 为 54.7%。使用无标记质谱法分析精液蛋白，并辅以统计和生物信息学分析。这种方法鉴定了 1445 种蛋白质，与蛋白质折叠、细胞间粘附、NADH 脱氢酶活性、ATP 结合、蛋白酶体复合物等过程有关。在 LF 和 HF 公牛中，有 338 种精液蛋白差异表达（p<0.05）。基于多元分析，BSP5 和精核核糖核酸酶定义了 HF 表型，而精子结合蛋白-1、凝胶蛋白、微管蛋白、甘油醛-3-磷酸脱氢酶、钙调蛋白、ATP 合酶、精子赤道段蛋白 1、过氧化物酶 5、分泌球蛋白家族 1D 和葡萄糖-6-磷酸异构酶则定义了 LF 表型。回归模型表明，公牛的 %ViableSperm 与精浆过氧化物酶 5、精子结合蛋白-1 和精子结合蛋白-1×BSP5 相互作用有关（R=0.84 和 0.79；p<0.05）。本报告是牛精液蛋白质组学最大的数据集。精液非细胞微环境的特定蛋白质可能是精子抗冻性的潜在标志物。
