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冰期耐力:冷冻保存对鲤鱼精子蛋白的影响

Ice-age endurance: the effects of cryopreservation on proteins of sperm of common carp, Cyprinus carpio L.

机构信息

University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, Research Institute of Fish Culture and Hydrobiology, Zatisi 728/II, 389 25 Vodnany, Czech Republic.

出版信息

Theriogenology. 2010 Aug;74(3):413-23. doi: 10.1016/j.theriogenology.2010.02.024. Epub 2010 Jun 8.

Abstract

Damage to spermatozoa during cryopreservation is regarded as a major obstacle to the expansion of sperm storage technology. The authors used two-dimensional polyacrylamide gel electrophoresis and matrix-associated laser desorption/ionization time-of-flight mass spectrometry to explore whether the protein profile of common carp (Cyprinus carpio) spermatozoa is affected by cryopreservation. Fourteen protein spots were significantly altered following cryopreservation. Eleven of these were identified: three as specific membrane proteins (N-ethylmaleimide-sensitive fusion protein attachment protein alpha, cofilin 2, and annexin A4) involved in membrane trafficking, organization, and cell movement; six as cytoplasmic enzymes (S-Adenosylhomocysteine hydrolase, Si:dkey-180p18.9 protein, lactate dehydrogenase B, phosphoglycerate kinase 1, transaldolase 1, and esterase D/formylglutathione hydrolase) involved in cell metabolism, oxidoreductase activity, and signal transduction; and two as transferrin variant C and F. Based on these findings, the authors hypothesize that transferrin in cryopreserved sperm may protect spermatozoa against oxidative damage during the freeze-thaw process. Cryopreservation caused changes in spermatozoa protein profiles that may lead to decreased spermatozoa velocity, motility, and fertilization success, and to subsequent ova hatching rate.

摘要

精子的冷冻损伤被认为是扩大精子储存技术的主要障碍。作者采用二维聚丙烯酰胺凝胶电泳和基质辅助激光解吸/电离飞行时间质谱技术,探讨了鲤鱼精子的蛋白质谱是否受到冷冻保存的影响。冷冻保存后,有 14 个蛋白质斑点发生明显改变。其中 11 个被鉴定为:3 个是参与膜运输、组织和细胞运动的特定膜蛋白(N-乙基马来酰亚胺敏感融合蛋白附着蛋白 α、丝切蛋白 2 和膜联蛋白 A4);6 个是细胞质酶(S-腺苷同型半胱氨酸水解酶、Si:dkey-180p18.9 蛋白、乳酸脱氢酶 B、磷酸甘油酸激酶 1、转醛醇酶 1 和酯酶 D/甲酰基谷胱甘肽水解酶),参与细胞代谢、氧化还原酶活性和信号转导;还有 2 个是转铁蛋白变体 C 和 F。基于这些发现,作者假设冷冻保存的精子中的转铁蛋白可能在冷冻和解冻过程中保护精子免受氧化损伤。冷冻保存导致精子蛋白质谱发生变化,可能导致精子速度、活力和受精成功率下降,以及随后的卵孵化率下降。

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