Usuga Alexandra, Rojano Benjamín A, Restrepo Giovanni
Faculty of Veterinary Medicine and Animal Science, CES University, Cl. 10 No. 22-04, 050021, Medellín, Colombia.
Faculty of Sciences, School of Chemistry, Universidad Nacional de Colombia, Cl. 59A No. 63-20, 050034, Medellín, Colombia.
Reprod Fertil Dev. 2018 Mar;30(3):563-569. doi: 10.1071/RD17044.
Contribution of seminal plasma proteins to semen freezability has been reported in several species, suggesting these proteins as genetic markers. The aim of this study was to evaluate the relationship between cysteine-rich secretory protein-3 (CRISP-3) and some of its single-nucleotide polymorphisms (SNPs) with post-thawing semen quality in stallions. DNA was obtained from 100 stallions, regions of interest were amplified by polymerase chain reaction and sequenced. Evaluated SNPs within the equine CRISP-3 gene were CRISP3c.+199A>G (SNP1), CRISP3c.+566C>A (SNP2), CRISP3c.+622G>A (SNP3) and CRISP3c.+716A>G (SNP4). CRISP-3 protein content in seminal plasma was determined by enzyme-linked immunosorbent assay. Semen from 30 stallions was cryopreserved and post-thaw motility, kinetics, abnormal morphology (AM), sperm vitality (SV) and membrane integrity (MI) were evaluated. Generalized linear models were fitted and means were compared using Tukey's test. Correlation and regression analyses were performed. For SNP1 and SNP3, the AA genotype had the highest results for motility and MI; for SNP2, the best results for motility and AM were obtained with the CC genotype. For SNP4, the GG genotype had the lowest results, except for MI. A high level of CRISP-3 protein in seminal plasma had the best results for motility, kinetics, SV and AM. In conclusion, there was a relationship between CRISP-3 genotype and seminal plasma protein and post-thawing semen quality in stallions.
在多个物种中都报道了精浆蛋白对精液冷冻保存能力的贡献,这表明这些蛋白可作为遗传标记。本研究的目的是评估富含半胱氨酸的分泌蛋白-3(CRISP-3)及其一些单核苷酸多态性(SNP)与种马解冻后精液质量之间的关系。从100匹种马中获取DNA,通过聚合酶链反应扩增感兴趣区域并进行测序。评估的马CRISP-3基因内的SNP为CRISP3c.+199A>G(SNP1)、CRISP3c.+566C>A(SNP2)、CRISP3c.+622G>A(SNP3)和CRISP3c.+716A>G(SNP4)。通过酶联免疫吸附测定法测定精浆中CRISP-3蛋白含量。对30匹种马的精液进行冷冻保存,并评估解冻后的活力、动力学、异常形态(AM)、精子活力(SV)和膜完整性(MI)。拟合广义线性模型,并使用Tukey检验比较均值。进行相关和回归分析。对于SNP1和SNP3,AA基因型在活力和MI方面的结果最高;对于SNP2,CC基因型在活力和AM方面获得了最佳结果。对于SNP4,除MI外,GG基因型的结果最低。精浆中高水平的CRISP-3蛋白在活力、动力学、SV和AM方面的结果最佳。总之,种马的CRISP-3基因型、精浆蛋白与解冻后精液质量之间存在关联。