Rego J P A, Martins J M, Wolf C A, van Tilburg M, Moreno F, Monteiro-Moreira A C, Moreira R A, Santos D O, Moura A A
J Anim Sci. 2016 Dec;94(12):5308-5320. doi: 10.2527/jas.2016-0811.
The objective of the present study was to describe the relationship of seminal plasma and total sperm cell proteins with the semen freezability parameters of Guzerat bulls. Thirteen bulls were subjected to breeding soundness evaluation. Semen samples were collected, cryopreserved, and then post-thawing sperm kinetics were assessed, where high ( = 7) and low ( = 6) freezability groups were defined. Seminal plasma and total sperm proteins from the 2 groups were separated by 2-dimensional SDS-PAGE, and spots were identified by mass spectrometry. Semen parameters post-cryopreservation were as follows in the high and low freezability groups, respectively: mean total motility, 52.4 ± 20.5 and 13.7 ± 3.9; percentage of normal sperm, 89.0 ± 2.6 and 64.7 ± 14.0; and reactivity of hypo-osmotic swelling test, 38.9 ± 4.7 and 13.6 ± 3.7. Three seminal plasma proteins (osteopontin-K, DNase γ precursor, and DNASE1L3) and 6 proteins from sperm cells (acrosome formation-associated factor isoform 2, annexin A1, disintegrin and metalloproteinase domain-containing protein 2, dihydrolipoyl dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase) were highly expressed ( < 0.05) in the high freezability group. Another 6 seminal plasma proteins (acrosin inhibitor 1, glutathione peroxidase 3, metalloproteinase inhibitor 2, ephrin-A1, annexin A1, and platelet-activating factor acetylhydrolase) were significantly higher ( < 0.05) in the low freezability group. We described the associations of seminal plasma and sperm cell proteins with post-thawing sperm viability of Guzerat bulls raised in a typical semiarid environment. Such associations indicate that specific seminal plasma proteins more abundant in bulls of low semen freezability may be a response to an early oxidative stress that is not detected by conventional prefreezing semen evaluation. Moreover, specific sperm proteins were more associated with good freezability. The results presented here can serve as guidelines for future research aiming to develop better extenders and/or to improve bull semen selection for cryopreservation.
本研究的目的是描述精液血浆和精子细胞总蛋白与古泽拉特公牛精液冷冻保存性能参数之间的关系。对13头公牛进行繁殖健全性评估。采集精液样本，进行冷冻保存，然后评估解冻后精子活力，据此定义了高（n = 7）、低（n = 6）冷冻保存性能组。通过二维SDS-PAGE分离两组的精液血浆和精子总蛋白，并用质谱法鉴定斑点。高、低冷冻保存性能组冷冻保存后的精液参数分别如下：平均总活力，52.4±20.5和13.7±3.9；正常精子百分比，89.0±2.6和64.7±14.0；低渗肿胀试验反应性，38.9±4.7和13.6±3.7。三种精液血浆蛋白（骨桥蛋白-K、DNaseγ前体和DNASE1L3）以及六种精子细胞蛋白（顶体形成相关因子亚型2、膜联蛋白A1、含解整合素和金属蛋白酶结构域蛋白2、二氢硫辛酰胺脱氢酶和甘油醛-3-磷酸脱氢酶）在高冷冻保存性能组中高表达（P < 0.05）。另外六种精液血浆蛋白（顶体蛋白酶抑制剂1、谷胱甘肽过氧化物酶3、金属蛋白酶抑制剂2、ephrin-A1、膜联蛋白A1和血小板活化因子乙酰水解酶）在低冷冻保存性能组中显著更高（P < 0.05）。我们描述了精液血浆和精子细胞蛋白与在典型半干旱环境中饲养的古泽拉特公牛解冻后精子活力的关联。这种关联表明，精液冷冻保存性能低的公牛中更丰富的特定精液血浆蛋白可能是对传统冷冻前精液评估未检测到的早期氧化应激的一种反应。此外，特定的精子蛋白与良好的冷冻保存性能更相关。本文给出的结果可为未来旨在开发更好的稀释液和/或改善公牛精液冷冻保存选择的研究提供指导。
