Characterization of capacitation, cryoinjury, and the role of seminal plasma in porcine sperm
Characterization of capacitation, cryoinjury, and the role of seminal plasma in porcine sperm
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DOI：
10.1016/j.theriogenology.2011.06.021
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发表时间：
2011-11-01
期刊：
影响因子：
2.8
通讯作者：
Althouse, Gary C.
中科院分区：
文献类型：
--
作者：
Vadnais, Melissa L.;Althouse, Gary C.
获能是精子必须经历的能够使卵母细胞受精的生化途径，而冷冻损伤是冷冻保存引起的生物物理损伤，其使精子立即能够受精。获能和冷冻损伤之间的相似性尚未完全阐明。本研究试图描述这两个过程，包括精浆（SP）的作用。Mercury-540染色检测到体外获能精子的质膜异常率为60.5%，而冷冻精子的质膜异常率为91.4%（P < 0.01），冷冻精子的质膜异常率为42.8%，体外获能精子的质膜异常率为13.6（P < 0.01），如通过膜联蛋白-V染色所评估的（SP减少两个群体中的磷脂酰丝氨酸倒位）。脂筏相关的糖脂GM，运动增加整个精子膜在冷冻保存的精子，虽然SP不影响这些精子的脂筏运动。冷冻保存和体外获能精子具有相似的酪氨酸磷酸化强度（尽管SP降低了这种强度）。在体外获能的精子中，67.5%的精子发生了离子载体诱导的顶体反应，而在冷冻保存的精子中有91.3%的精子发生了顶体反应。在这两种情况下，SP使顶体反应精子的百分比分别降低（P < 0.01）至1.0%和7.8%。冷冻保存似乎会损害精子，导致膜紊乱、胆固醇流出和获能精子百分比显著增加。在获能和冷冻损伤的精子，除了SP出现减弱这些事件中的一些。(C)2011 Elsevier Inc. All rights reserved.
Capacitation is a biochemical pathway sperm must undergo to be able to fertilize an oocyte, whereas cryoinjury is cryopreservation-induced biophysical damage which renders sperm immediately capable of fertilization. Similarities between capacitation and cryoinjury have not been fully elucidated. The present study attempted to characterize both processes, including the role of seminal plasma (SP). Merocyanine-540 staining detected an increase (P < 0.01) in plasma membrane disorder from 60.5% in in vitro capacitated sperm to 91.4% in cryopreserved sperm, with no effect of SP. After cryopreservation, 42.8% of sperm displayed phosphatidylserine on the outer leaflet compared to 13.6% of in vitro capacitated sperm (P < 0.01), as assessed by annexin-V staining (SP decreased phosphatidylserine inversion in both populations). Lipid raft-associated glycolipid GM, movement increased throughout the entire sperm membrane in cryopreserved sperm, although SP did not affect lipid raft movement in these sperm. Cryopreserved and in vitro capacitated sperm had a similar intensity of tyrosine phosphorylation (although SP reduced this intensity). In in vitro capacitated sperm, 67.5% underwent an ionophore induced acrosome reaction with 91.3% reacting in cryopreserved sperm. In both cases, SP reduced (P < 0.01) the percentage of acrosome-reacted sperm to 1.0 and 7.8%, respectively. Cryopreservation appeared to damage sperm, resulting in marked increases in membrane disorder, cholesterol efflux, and percent of capacitated sperm. In both capacitated and cryoinjured sperm, the addition of SP appeared to attenuate some of these events. (C) 2011 Elsevier Inc. All rights reserved.