Valencia Julián, Gómez Germán, López Walter, Mesa Henry, Henao Francisco Javier
Universidad de Caldas, Faculty of Agricultural Sciences, A.A. 275, Manizales, Caldas Colombia.
J Anim Sci Biotechnol. 2017 Mar 1;8:21. doi: 10.1186/s40104-017-0151-y. eCollection 2017.
The purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen: the heat shock protein 90 alpha (HSP90a), the Niemann-Pick disease type C2 protein (NPC2), and lipocalin-type prostaglandin D synthase (L-PGDS). Six adult boars (each boar was ejaculated three times, 18 in total) were classified by freezability based on the percentage of functionally competent sperm. The male semen with highest freezability (MHF) and the male semen with lowest freezability (MLF) were centrifuged immediately after collection to separate seminal plasma and spermatozoa to make four possible combinations of these two components and to incubate them for 3 h, adjusting the temperature to 17 °C, to freeze them afterwards. The quantification of proteins was performed in two stages: at zero and at 3 h after incubation of the four combinations.
The spermatozoa × incubation time (IT) interaction only had effect ( < 0.01) on HSP90a levels; this protein increased in seminal plasma, after 3 h of incubation, in larger quantity ( < 0.05) in combinations with MLF spermatozoa. In relation with the NPC2 protein, two isoforms of 16 and 19 kDa were identified. The 19 kDa isoform was affected ( < 0.01) only by the seminal plasma × IT interaction, with superior values ( < 0.01) both at zero and three hours of incubation, in the combinations with MHF seminal plasma; and 16 kDa isoform was affected ( < 0.01) only by the IT with reduction after 3 h of incubation. The levels of L-PGDS was affected ( < 0.01) only by the spermatozoa × IT interaction, which reduced ( < 0.01) in combinations with MLF spermatozoa after 3 h of incubation.
It is possible to consider that the three proteins evaluated were associated with freezability of boar semen due, especially, to the fact that mixtures with MLF spermatozoa showed greater increase levels of the HSP90a protein and reduction of L-PGDS in plasma. In addition, the seminal plasma of MHF had higher concentration of the NPC2 of 19 kDa protein, which was reduced by incubating with MHF spermatozoa.
本研究旨在确定参与精子功能的三种蛋白质与猪精液冷冻能力之间的关联，这三种蛋白质分别是热休克蛋白90α（HSP90a）、尼曼-匹克病C2型蛋白（NPC2）和脂钙素型前列腺素D合成酶（L-PGDS）。根据功能正常精子的百分比，将6头成年公猪（每头公猪采精3次，共18次）按冷冻能力进行分类。采集后立即对冷冻能力最高的公猪精液（MHF）和冷冻能力最低的公猪精液（MLF）进行离心，分离精浆和精子，形成这两种成分的四种可能组合，并将它们在17℃下孵育3小时，之后进行冷冻。蛋白质定量分两个阶段进行：在四种组合孵育0小时和3小时时。
精子×孵育时间（IT）的相互作用仅对HSP90a水平有影响（P＜0.01）；孵育3小时后，与MLF精子组合的精浆中，这种蛋白质的含量增加得更多（P＜0.05）。关于NPC2蛋白，鉴定出16 kDa和19 kDa两种亚型。19 kDa亚型仅受精浆×IT相互作用的影响（P＜0.01），在与MHF精浆组合孵育0小时和3小时时，其值均较高（P＜0.01）；16 kDa亚型仅受IT的影响（P＜0.01），孵育3小时后含量降低。L-PGDS水平仅受精子×IT相互作用的影响（P＜0.01），孵育3小时后，与MLF精子组合时其水平降低（P＜0.01）。
可以认为，所评估的这三种蛋白质与公猪精液的冷冻能力有关，特别是因为与MLF精子的混合物显示HSP90a蛋白水平有更大的升高，而血浆中L-PGDS水平降低。此外，MHF的精浆中19 kDa的NPC2蛋白浓度较高，与MHF精子孵育后其浓度降低。
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