Hernández Marta, Roca Jordi, Calvete Juan J, Sanz Libia, Muiño-Blanco Teresa, Cebrián-Pérez José A, Vázquez Juan M, Martínez Emilio A
Departamento de Medicina y Cirugía Animal, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, 30071 Murcia, Spain.
J Androl. 2007 Sep-Oct;28(5):689-97. doi: 10.2164/jandrol.107.002725. Epub 2007 Apr 25.
The study evaluated the protective effect of seminal plasma (SP) added to freezing extender against cryopreservation injuries to boar spermatozoa. Pooled sperm-rich fractions collected from 9 fertile boars were frozen in 0.5-mL straws after being extended in a conventional freezing extender either alone or supplemented with 5% of SPs (SP1-SP4) collected from the sperm-rich fractions (diluted 1:1, vol/vol, in Beltsville Thawing Solution extender) from 4 boars (1-4) with known sperm cryosurvival (poor, moderate, and good sperm freezers). Cryopreservation injuries were assessed in terms of postthaw sperm motility (assessed by computer-assisted sperm analysis), viability (plasma membrane and acrosome integrity assessed simultaneously by flow cytometry), membrane lipid peroxidation (malondialdehyde [MDA] production), and the ability of thawed spermatozoa to fertilize in vitro-matured homologous oocytes. The addition of SP from good sperm freezers (SP3 and SP4) improved (P < .01) the motility and viability of thawed spermatozoa without any influence on MDA production. Moreover, SP from good sperm freezers also increased (P < .05) the percentage of penetrated (SP3) and polyspermic oocytes (SP4) with respect to the control. Neither the total amount of SP proteins, protein profiles, nor antioxidant capacity of the different SPs were related to the various cryosurvival/fertilizing capacities of the processed spermatozoa.
本研究评估了添加到冷冻稀释液中的精浆(SP)对猪精子冷冻保存损伤的保护作用。从9头可育公猪收集的富含精子的混合部分,在单独使用传统冷冻稀释液或添加从4头已知精子冷冻存活率(精子冷冻效果差、中等和良好)的公猪(1 - 4)的富含精子部分收集的5% SP(SP1 - SP4)(在Beltsville解冻液稀释液中按1:1体积/体积稀释)中进行稀释后,以0.5 mL细管冷冻保存。根据解冻后精子活力(通过计算机辅助精子分析评估)、存活率(通过流式细胞术同时评估质膜和顶体完整性)、膜脂质过氧化(丙二醛[MDA]生成)以及解冻精子使体外成熟的同源卵母细胞受精的能力来评估冷冻保存损伤。添加来自精子冷冻效果良好的公猪的SP(SP3和SP4)可提高(P <.01)解冻后精子的活力和存活率,且对MDA生成无任何影响。此外,来自精子冷冻效果良好的公猪的SP相对于对照组也增加了(P <.05)穿透卵母细胞(SP3)和多精入卵卵母细胞(SP4)的百分比。不同SP的SP蛋白总量、蛋白谱或抗氧化能力均与处理后精子的各种冷冻存活/受精能力无关。