海藻糖对猪精子冷冻真空干燥保存效果的影响

Effect of trehalose on the freeze-dried boar spermatozoa

摘要:

猪精子经冷冻干燥后，在光学显微镜和电子显微镜下观察其超微结构，并借助辅助生殖技术将其注入猪卵母细胞后，进一步观察受精卵的发育情况。结果表明：海藻糖组雄原核形成率 (68.52％)、卵裂率 (59.17％) 和囊胚率 (19.16％) 优于EDTA组 (64.59％、56.26％和15.62％) 和对照组 (35.36％、52.33％和8.60％) (P＜0.05)；海藻糖组的冷冻真空干燥猪精子分别在4℃下保存60、120、180 d，雄原核形成率、卵裂率和囊胚率均无显著差异 (P＞0.05)；海藻糖组的冷冻真空干燥猪精子复水化后孵育1 h和2 h，卵裂率、卵裂率和囊胚率均差异显著 (P＜0.05)；海藻糖处理组与EDTA处理组中的冷冻真空干燥猪精子分别在4℃和?20℃下保存后各处理组间精子形态差异不显著 (P＞0.05)；海藻糖组中B级冷冻真空干燥精子百分数显著多于EDTA处理组 (P＜0.05)。超微结构分析表明，冷冻真空干燥猪精子的损伤主要表现在顶体和颈部的肿胀与缺损、尾部断裂。

Abstract:

After freeze-drying, the ultrastructure of boar sperms was observed by optical and electron microscopy. The in vitro development ability of the sperm was also examined with intracytoplasmic sperm injection (ICSI). The rate of male pronuclear formation was (68.52%), for cleavage (59.17%) and for blastocyst formation (19.16%) of the trehalose group (0.2 mol/L), significantly higher than those of the 50 mmol/L EDTA group (64.59%, 56.26% and 15.62%) and the control group (35.36%, 52.33% and 8.60%) (P<0.05). After storage for 60, 120 and 180 d at 4°C, no significant difference in the in vitro development was observed (P>0.05). The male pronuclear, cleavage and blastocyst formation after ICSI with freeze-dried spermatozoa incubated for 1 h was superior than those incubated for 2 h (P<0.05). No significant differences in the structures after stored at 4°C or ?20°C (P>0.05) were observed between the trehalose group and EDTA group. The percent of B grade freeze-dried boar spermatozoa in the trehalose group was higher than that of the EDTA group (P<0.05). Based on the ultrastructure observation, main cryogenic damage in freeze-dried boar spermatozoa was swelling, damage or rupture in the sperm acrosome, neck and tail.