孕马血清促性腺激素对狗卵母细胞体外核成熟的影响  

Effects of PMSG on Maturation of Canine Oocytes

作　　者：王坤[1] 林涛[1] 赵明辉[1] 高威威[1] 尹熙俊[1] 

机构地区：[1]延边大学农学院动物科学系,吉林龙井133400

出　　处：《安徽农业科学》2008年第35期15503-15504,15509,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]研究孕马血清促性腺激素(PMSG;eCG)对狗卵母细胞体外成熟效果的影响。[方法]采用切割法收集卵巢表面卵丘卵母细胞复合体(COCs),并且随机分为4组,在添加不同浓度的PMSG的基础培养液(含有10%胎牛血清的DMEM)中,5%CO2,38.5℃培养箱内培养48 h。在含有0.1%透明质酸酶的D-PBS中用细小玻璃管剥离卵丘细胞从而获得裸卵,为了鉴别核成熟期,用4%甲醛溶液固定裸卵15 min,然后用10μg/ml Hoechst33342染色、压片,荧光显微镜下观察核形态。[结果]体外培养48 h后,添加PMSG处理组卵丘扩散明显,但发育到MⅠ-MⅡ的比率都没有显著差异(P(0.05)。100 IU/ml PMSG处理组GVBD期率显著低于对照组。[结论]添加PMSG改善了卵丘扩散的效果,但是没有提高核成熟率,需进一步改进狗卵母细胞体外成熟率的培养体系。[ Objective ] The aim of this study was to evaluate the effects of PMSG on nuclear maturation of canine oocyte in vitro. [ Method ] Cumulus-ooeyte complexes （COCs） were released by slicing. They were randomly divided into four groups and cultured in DMEM containing 10% foetal bovine serum （FBS） supplemented with different concentrations of PMSG at 38.5 ℃, 5% CO2 in air for 48 h. To evaluate the nuclear meiotic stage, oocytes were denuded by placing them in D-PBS supplemented with 0.1% hyaluronidase, and then gently pipetted using small glass pipettes. Denuded oocytes were fixed with 4% formaldehyde for 15 rain and 10 μg/ml Hoeehst33342 then mounted on the slides to evaluate nuclear morphology using a microscope equipped with fluorescent illumination. [ Result] PMSG supplemented with the cumulus expansion was better. However, the percentages of MI-M 11 in PMSG groups had no significant differences （P 〈 0.05 ）. GVBD of 100 IU/ml PMSG group was much lower than that in the control group. [ Conclusion] PMSG supplement in IVM medium could increase cumulus cell expansion, while the nuclear maturation rate remained low. Further studies were required to improve the nuclear development rate of canine oocytes.

关 键 词：狗 卵母细胞 核成熟 孕马血清促性腺激素 

分 类 号：S829.9[农业科学—畜牧学]