不同诱导剂对山羊精子体外获能和受精能力的影响

Capacitation and Fertilization of Goat Spermatozoa Treated with Different Capacited Inducers in vitro

比较不同获能诱导剂对山羊精子体外获能及受精能力的影响,优化山羊早期胚胎体外生产技术体系.山羊精子分别以不同浓度肝素、IA和EGS处理后,检查精子获能率、活力和体外存活时间,同时与体外成熟的卵母细胞受精,观察不同获能处理对精子受精能力的影响.在培养液中添加5,10,20 μg/mL肝素,都能促进精子获能,其中10μg/mL组的获能率达到55.6%,存活时间为18～19 h;低于此浓度,获能率显著降低,高于此浓度,获能率升高不显著,但精子存活时间和活力明显下降.0.1,0.4,0.7μmol/LIA都能提高精子获能率,其中0.4和0.7 μmol/L处理组精子获能率最高,分别为56.2%和53.6%,但两组中的精子活力和存活时间有明显差异.5%,10%和20%EGS都能提高精子活力,延长体外存活时间,但实验组的获能率最高不到20%.10%EGS获能组的受精率和卵裂率分别为27.9%和18.6%,显著低于10μg/mL肝素和0.4μmoL/LIA组,后两者的受精率相似,但IA组的卵裂率明显高于肝素组(52.0%VS 40.2%,P＜0.05).10μg/mL肝素和0.4 μmol/L IA都可诱导山羊精子高水平获能,并获得较好的受精能力,但IA表现出更强的促卵裂发生效应.

To optimize the technological system for goat embryo production in vitro, capacitation and fertilization of goat spermatozoa were compared after treated with different capacited inducers. After treated with different dosage of heparin, Calcium ionophore A23187 （IA） and estrus goat serum （EGS）, the motility , capacitated percentage, longevity and fertilizing ability of goat spermatozoa were assessed. The results showed that when spermatozoa was capacited with 5, 10,20μg/mL heparin, capacitated percentages and longevities were 55.6% and 18 - 19 h in 10μg/mL group. There were lower proportion of capacitation in 5μg/mL group and lower motility and shorter longevity in 20μg/mL group, compared with 10μg/mL group. When spermatozoa treated with 0.1,0.4,0.7μmol/L IA, the highest capacitated percentages of 53.8% and 56.2% were appeared in 0.4 and 0.7μmol/L groups, respectively. However, motilities and longevities in both groups showed significant differences. Capacitated percentages were less than 20 % when spermatozoa were treated with 5%, 10% and 20% EGS, although motilities and longevities were promoted in these groups. The lowest rates of fertilization and cleavage was appeared in 10% EGS group （27.9% and 18.6% ）. Rate of fertilization in 0.4mol/L IA group was similar to 10 g/mL heparin group （70.7% and 68.2% ）, but cleavage rate of the former was signif icantly higher than that of the latter （52.0 % VS 40.2 % ; P 〈 0.05）. Higher Capacitated percentages and fertilized abilities of goat spermatozoa are appeared after treated with 10 g/mL heparin and 0.4 mol/L IA, and IA promote cleavage rate after fertilization.