Membranous and structural damage that occur during cryopreservation of human sperm may be time-related events
Membranous and structural damage that occur during cryopreservation of human sperm may be time-related events
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DOI：
10.1016/j.fertnstert.2005.11.046
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发表时间：
2006-06-01
影响因子：
6.7
通讯作者：
Sullivan, Robert
中科院分区：
文献类型：
--
作者：
Desrosiers, Pascal;Legare, Christine;Sullivan, Robert
目的：用精质膜蛋白P34H和α -微管蛋白作为亚细胞区隔标志物，评价精子低温保存损伤。设计：前瞻性实验研究。环境：学术医院研究中心和生育诊所。患者：从到生育诊所就诊的健康捐赠者处取得的精液样本。精子样品直接进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和Western blot实验（对照组）或在液氮中低温保存不同时间后进行分析。干预措施：SDS-PAGE， Western blotting，以及冷冻前后P34H和α -微管蛋白的密度定量。主要观察指标：精子冷冻保存后各组间蛋白质含量的变化。结果：在31个用于P34H评估的精子样本中，与对照组相比，精子冷冻保存后减少了50%。41个精子样本的α微管蛋白免疫印迹显示，与对照组相比，冷冻保存精子组的蛋白质检测增加了200%。与P34H检测相反，免疫检测到的α -微管蛋白水平的变化似乎与冷冻保存时间有关，因为它在储存期间增加。结论：这些研究结果表明，人类精液冷冻保存在不同的细胞水平上引起损伤。此外，一些低温损伤是立即发生的，而另一些似乎是在液氮中储存的一段时间内发生的。
Objective: To evaluate sperm cryopreservation-induced injuries using sperm plasma membrane protein P34H and alpha-tubulin as two different subcellular compartment markers.Design: Prospective experimental study.Setting: Academic hospital research center and fertility clinic.Patient(s): Semen samples obtained from healthy donors attending the fertility clinic. Sperm samples were either directly processed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot experiments (control group) or cryopreserved in liquid nitrogen for different periods of time before being analyzed.Intervention(s): SDS-PAGE, Western blotting, and densitometric quantification of P34H and alpha-tubulin before and after cryopreservation.Main Outcome Measure(s): Changes in protein quantification between the different groups as a result of sperm cryopreservation.Result(s): In the 31 sperm samples processed for P34H evaluation, a 50% decrease is observed after sperm cryopreservation as compared to the control group. The alpha-tubulin immunoblotting of 41 sperm samples revealed a 200% increase in the protein detection in the group of cryopreserved sperm as compared to the control group. Contrary to the P34H detection, this change in alpha-tubulin immunodetected levels appears to be related to the cryopreservation period as it increases during storage.Conclusion(s): These findings indicate that cryopreservation of human semen induces damages at different cellular levels. Moreover, some cryoinjuries are immediate although others seem to take place over time stored in liquid nitrogen.