Relationship between HSP90a, NPC2 and L-PGDS proteins to boar semen freezability

Julián Valencia; Germán Gómez; Walter López; Henry Mesa; Francisco Javier Henao

摘要

Background: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 (HSPg0a),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 ℃,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.Results:The spermatozoa × incubation time (IT) interaction only had effect (P ＜ 0.01) on HSPg0a levels;this protein increased in seminal plasma,after 3 h of incubation,in larger quantity (P ＜ 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 (P ＜ 0.01) only by the seminal plasma × IT interaction,with superior values (P ＜ 0.01) both at zero and three hours of incubation,in the combinations with MHF seminal plasma;and 16 kDa isoform was affected (P＜ 0.01) only by the IT with reduction after 3 h of incubation.The levels of L-PGDS was affected (P＜ 0.01)only by the spermatozoa × IT interaction,which reduced (P＜ 0.01) in combinations with MLF spermatozoa after 3 h of incubation.Conclusions: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.

机译：背景：本研究的目的是确定与精子功能有关的三种蛋白质与猪精液可冻结性的关系：热休克蛋白90α（HSPg0a），尼曼-匹克病C2型蛋白（NPC2）和脂蛋白型前列腺素D合酶（L-PGDS）。根据功能性精子的百分比，将6只成年公猪（每头公猪射出3次，共18只）按可冷冻性分类。可冷冻性最高的雄性精液（MHF）和收集后立即将具有最低冷冻性能（MLF）的雄性精液离心，以分离精浆和精子，以使这两种成分有四种可能的组合，并将它们孵育3小时，将温度调节至17℃，然后冷冻。四种组合的孵育分别在零和3 h的两个阶段进行。结果：精子×孵育时间（IT）相互作用仅对HSPg0l有影响（P ＜0.01）。卵泡;该蛋白在精浆中孵育3 h后增加，与MLF精子合用时量更大（P ＜0.05）。与NPC2蛋白相关，鉴定出16和19 kDa的两个亚型。19kDa的亚型。与MHF精浆联合使用，仅受精浆×IT相互作用的影响（P ＜0.01），在孵化零和3小时时均具有较高的值（P ＜0.01）;并影响16 kDa亚型（P <0.01）。孵育3 h后IT降低（P <0.01）。仅L.PGDS水平受精子×IT相互作用的影响（P <0.01），与MLF精子组合3 h降低（P <0.01）。结论：有可能认为所评估的三种蛋白质与公猪精液的可冻结性有关，特别是由于与MLF精子的混合物显示出血浆中HSP90a蛋白水平的增加更大，而L-PGDS的减少此外，MHF的精浆具有NPC2的19 kDa蛋白质浓度较高，但与MHF精子孵育后降低了浓度。

Relationship between HSP90a, NPC2 and L-PGDS proteins to boar semen freezability

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