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热应激期间,HSP90维持公猪精子活力和线粒体膜电位。

HSP90 maintains boar spermatozoa motility and mitochondrial membrane potential during heat stress.

作者信息

Calle-Guisado V, Bragado M J, García-Marín L J, González-Fernández L

机构信息

Research Group of Intracellular Signalling and Technology of Reproduction (SINTREP), Institute of Biotechnology in Agriculture and Livestock (INBIO G+C), University of Extremadura, Cáceres, Spain.

Research Group of Intracellular Signalling and Technology of Reproduction (SINTREP), Institute of Biotechnology in Agriculture and Livestock (INBIO G+C), University of Extremadura, Cáceres, Spain.

出版信息

Anim Reprod Sci. 2017 Dec;187:13-19. doi: 10.1016/j.anireprosci.2017.09.009. Epub 2017 Sep 29.

Abstract

Heat Shock Proteins (HSP) is a family of proteins that protects cells from high temperatures. The present work aimed to elucidate the role that HSP90 exerts on boar sperm incubated under heat stress conditions on viability, total motility (TM), progressive motility (PM), acrosome status, mitochondrial membrane potential and plasma membrane lipid organization. Sperm were incubated in non-capacitating conditions (Tyrode's basal medium or TBM) for 3, 8 and 24h or in capacitating conditions (Tyrode's complete medium or TCM) for 4h at 38.5°C or 40°C (Heat stress) in the presence or absence of 5 or 20μM of 17-AAG, a specific HSP90 inhibitor. Sperm viability was not affected by the presence of 17-AAG in any condition tested compared with its own control (at the same temperature and incubation time). In non-capacitating conditions TM (22.7±4.1 vs. 1.9±1.1; % mean±SEM), PM (3.1±0.9 vs. 0) and high mitochondrial membrane potential (19.5±2.2 vs. 11.8±0.8) decreased significantly in sperm incubated at 40°C for 24h in the presence of 20μM 17-AAG (control vs. 20μM 17-AAG, respectively; p<0.05). In sperm incubated at 38.5°C only a mild decrease in TM was observed (48.7±3.1 vs. 32.1±4.8; control vs. 20μM 17-AAG, respectively; p<0.05). However, under capacitating conditions none of the sperm parameters studied were affected by 17-AAG after 4h of incubation. These results demonstrate for the first time the role of HSP90 in the maintenance of boar sperm motility and mitochondrial membrane potential during prolonged heat stress in non-capacitating conditions.

摘要

热休克蛋白(HSP)是一类保护细胞免受高温影响的蛋白质家族。本研究旨在阐明HSP90在热应激条件下孵育的公猪精子的活力、总运动率(TM)、前向运动率(PM)、顶体状态、线粒体膜电位和质膜脂质组织方面所发挥的作用。精子在非获能条件下(Tyrode基础培养基或TBM)于38.5°C或40°C(热应激)下孵育3、8和24小时,或在获能条件下(Tyrode完全培养基或TCM)于38.5°C或40°C(热应激)下孵育4小时,孵育过程中存在或不存在5或20μM的17-AAG(一种特异性HSP90抑制剂)。与各自的对照(在相同温度和孵育时间下)相比,在任何测试条件下,17-AAG的存在均未影响精子活力。在非获能条件下,在20μM 17-AAG存在下于40°C孵育24小时的精子中,TM(22.7±4.1对1.9±1.1;平均百分比±标准误)、PM(3.1±0.9对0)和高线粒体膜电位(19.5±2.2对11.8±0.8)显著降低(分别为对照对20μM 17-AAG;p<0.05)。在38.5°C孵育的精子中,仅观察到TM轻度下降(48.7±3.1对32.1±4.8;分别为对照对20μM 17-AAG;p<0.05)。然而,在获能条件下,孵育4小时后,所研究的精子参数均未受17-AAG影响。这些结果首次证明了HSP90在非获能条件下长时间热应激期间对公猪精子活力和线粒体膜电位维持中的作用。

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