海藻糖對(duì)豬精子冷凍真空干燥保存效果的影響

孟祥黔，顧曉龍，吳彩鳳，戴建軍，張廷宇，謝一妮，吳志強(qiáng)，劉亮，馬恒東，張德福

1 四川農(nóng)業(yè)大學(xué)生命科學(xué)與理學(xué)院，雅安 625014 2 上海農(nóng)業(yè)科學(xué)院畜牧獸醫(yī)研究所，上海 201106 3 上海農(nóng)業(yè)遺傳育種重點(diǎn)實(shí)驗(yàn)室 動(dòng)物遺傳工程研究室，上海 201106

海藻糖對(duì)豬精子冷凍真空干燥保存效果的影響

孟祥黔1,2,3，顧曉龍1,2,3，吳彩鳳2,3，戴建軍2,3，張廷宇2,3，謝一妮1,2,3，吳志強(qiáng)1,2,3，劉亮2,3，馬恒東1，張德福2,3

1 四川農(nóng)業(yè)大學(xué)生命科學(xué)與理學(xué)院，雅安 625014 2 上海農(nóng)業(yè)科學(xué)院畜牧獸醫(yī)研究所，上海 201106 3 上海農(nóng)業(yè)遺傳育種重點(diǎn)實(shí)驗(yàn)室 動(dòng)物遺傳工程研究室，上海 201106

豬精子經(jīng)冷凍干燥后，在光學(xué)顯微鏡和電子顯微鏡下觀察其超微結(jié)構(gòu)，并借助輔助生殖技術(shù)將其注入豬卵母細(xì)胞后，進(jìn)一步觀察受精卵的發(fā)育情況。結(jié)果表明：海藻糖組雄原核形成率(68.52%)、卵裂率(59.17%)和囊胚率(19.16%)優(yōu)于EDTA組(64.59%、56.26%和15.62%)和對(duì)照組(35.36%、52.33%和8.60%)(P＜0.05)；海藻糖組的冷凍真空干燥豬精子分別在4℃下保存60、120、180 d，雄原核形成率、卵裂率和囊胚率均無(wú)顯著差異(P＞0.05)；海藻糖組的冷凍真空干燥豬精子復(fù)水化后孵育1 h和2 h，卵裂率、卵裂率和囊胚率均差異顯著(P＜0.05)；海藻糖處理組與 EDTA處理組中的冷凍真空干燥豬精子分別在 4℃和?20℃下保存后各處理組間精子形態(tài)差異不顯著(P＞0.05)；海藻糖組中B級(jí)冷凍真空干燥精子百分?jǐn)?shù)顯著多于EDTA處理組(P＜0.05)。超微結(jié)構(gòu)分析表明，冷凍真空干燥豬精子的損傷主要表現(xiàn)在頂體和頸部的腫脹與缺損、尾部斷裂。

豬精子，冷凍真空干燥，單精子注射

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.

Keywords:boar spermatozoa, freeze-drying, intracytoplasmic sperm injection

早在1949年P(guān)olge等[1]首次發(fā)現(xiàn)甘油對(duì)精子具有冷凍保護(hù)作用，同時(shí)也進(jìn)行了精子冷凍真空干燥的試驗(yàn)。……