The protective effect of a 17 °C holding time on boar sperm plasma membrane fluidity after exposure to 5 °C
The protective effect of a 17 °C holding time on boar sperm plasma membrane fluidity after exposure to 5 °C
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DOI：
10.1016/j.cryobiol.2012.11.006
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发表时间：
2013-02-01
期刊：
影响因子：
2.7
通讯作者：
Althouse, G. C.
中科院分区：
文献类型：
--
作者：
Casas, I.;Althouse, G. C.
保持时间（HT）是指在进一步处理冷藏之前，将原始或稀释状态的射精保持在17摄氏度的时间。在公猪中，HT对精液冷却至15至5 ℃（质膜重塑发生的温度范围）的精子质量参数产生积极影响。HT对质膜结构的影响的客观见解仍然是未知的。因此，目前的工作试图阐明，如果HT有助于减少公猪精子质膜流动性的改变在冷冻保存实践中的冷却过程的初始步骤（保持在5 ℃），并与选择精子质量参数。从5头公猪收集19份精液，并根据不同的处理进行处理：T1）新鲜稀释精液，17 ℃下0 h; T2）新鲜稀释精液，17 ℃下24 h（HT）; 13）来自T1的精子在乳糖-蛋黄（LEY）稀释液中，5 ℃下3 h; 14）来自T2的精子在LEY中，5 ℃下3 h; T5）来自LEY中T1的精子，5 ℃下24 h; T6）来自LEY中T2的精子，5 ℃下24 h。使用CASA评估精子活力，并通过双标记流式细胞术（M540/YO-PRO（R）-1）评估精子质膜完整性和流动性。结果表明，与HT（T6）相比，缺乏HT（T5）暴露导致样本活动性降低，暴露于HT的精子表现出更低的质膜流动性。总的来说，这些结果提供了经验证据，证明在精液加工中加入HT可以通过维持质膜的脂质结构来保护野猪精子免受冷损伤。(C)2012 Elsevier Inc. All rights reserved.
The holding time (HT) is the period during which an ejaculate, either in a raw or diluted state, is held at 17 degrees C before further processing for cold-storage. In boars, the HT positively influences select sperm quality parameters of semen cooled from 15 to 5 degrees C, a range in temperature during which plasma membrane remodeling occurs. Objective insight into the effect of HT on plasma membrane organization remains unknown. Therefore, the present work sought to elucidate if HT contributes to minimizing alterations in boar sperm plasma membrane fluidity at the initial step of the cooling process in a cryopreservation practice (holding at 5 degrees C) and in relation with select sperm quality parameters. Nineteen ejaculates from five boars were collected and processed according to different treatments: T1) Fresh diluted semen, 0 h at 17 degrees C; T2) Fresh diluted semen, 24 h at 17 degrees C (HT); 13) Sperm from T1 in a lactose-egg yolk (LEY) extender, 3 h at 5 degrees C; 14) Sperm from T2 in LEY, 3 h at 5 degrees C; T5) Sperm from T1 in LEY, 24 h at 5 degrees C; T6) Sperm from T2 in LEY, 24 h at 5 degrees C. Sperm motility was assessed using CASA, and sperm plasma membrane integrity and fluidity were evaluated by flow cytometry with dual labeling (M540/YO-PRO (R)-1). Results demonstrated that the lack of exposure to a HT (T5) results in reduced sample motility compared to those having a HT (T6), with sperm exposed to HT exhibiting less plasma membrane fluidity. Collectively, these results provide empirical evidence that incorporation of a HT in semen processing protects boar sperm against cold injury through maintenance of lipid architecture of the plasma membrane. (C) 2012 Elsevier Inc. All rights reserved.