水牛GV期和MⅡ卵母细胞中脂肪含量及分布模式研究  

Study on Content and Distribution of Lipids of Buffalo Oocytes at GV and MⅡ Stage

作　　者：许惠艳 张筱芫[1,2] 杨小淦 卢晟盛 梁兴伟[1,2] 陆阳清 张明[1,2] 卢克焕 XU Hui-yan;ZHANG Xiao-yuan;YANG Xiao-gan;LU Sheng-sheng;LIANG Xing-wei;LU Yang-qing;Zhang Ming;LU Ke-huan(State Key Laboratory for Conset-eation and Utilization of Subtropical Agro-bioresources,Guangxi Nanning 530005,China;College of Animal Science ＆ Technology,Guangxi University,Guangxi Nanning 530005,China)

机构地区：[1]亚热带农业生物资源保护与利用国家重点实验室,广西南宁530005 [2]广西大学动物科学技术学院,广西南宁530005

出　　处：《西南农业学报》2018年第11期2413-2417,共5页Southwest China Journal of Agricultural Sciences

基　　金：广西自然科学基金(2018GXNSFAA138142);广西大学科研基金(XJZ170377);广西大学2017年实验室建设与实验教学改革项目(201711)

摘　　要：【目的】探讨水牛GV期未成熟卵和MⅡ期成熟卵母细胞中脂肪的含量及分布状态,为研究脂肪在水牛卵母细胞体外成熟过程中的作用及卵母细胞体外成熟体系的优化提供参考。【方法】从南宁市某屠宰场收集水牛卵巢并采集卵丘卵母细胞复合体,其中一部分复合体用于收集GV期未成熟卵母细胞,另一部分经体外成熟22～24 h后,获得MⅡ期成熟卵母细胞。之后把GV期和MⅡ期的卵母细胞放入含有10μg/m L Nile red的染色液中,室温黑暗环境中染色4～5 h后,在荧光显微镜下观察,拍照。根据所得照片观察卵母细胞中脂肪分布模式,同时借助Image J软件计算荧光强度,以荧光强度来计算卵母细胞中脂肪含量。【结果】MⅡ期卵子中脂肪含量显著低于GV期脂肪含量(66. 63±1. 197 vs. 70. 81±1. 328,P <0. 05)。GV期和MⅡ期卵母细胞中脂肪各类分布模式没有统计学上的差异(外围分布:3. 148±2. 298 vs. 5. 342±3. 858;半均匀分布:30. 85±4. 042 vs. 35. 70±4. 265;均匀分布:66. 00±3. 257 vs. 60. 60±3. 933,P> 0. 05)。【结论】从GV期未成熟卵母细胞到成熟MⅡ期卵母细胞成熟过程中,水牛卵子内脂肪含量显著降低,而卵母细胞成熟前后脂肪分布模式没有发生改变,多数以均匀分布模式为主。[ Objective] In order to explore the role of lipid in buffalo oocytes in vitro maturation process, and optimize the oocytes mature sys- tem, the lipid content and distribution of buffalo mature stage GV immature and MⅡ oocytes were investigated. [ Method] The ovaries of buf- falo were collected from a slaughterhouse in Nanning city to collect the cunmlus oocyte complexes, one part of the complexes were used to collect immature oocytes in GV stage, the MⅡ mature ooeytes were obtained from another part after in vitro maturation for 22 - 24 hours. Af- ter that, the oocytes of GV and MⅡ phases were placed into 10 μg/mL Nile red stain solution. After 4 -5 hours of staining in a dark envi- ronment at room temperature, oocytes were observed under a fluorescence microscope. According to the obtained photos, the lipid distribu- tion pattern in oocytes was observed, and the fluorescence intensity was calculated with the analysis of hnage J software, and the lipid content in oocytes was calculated with the fluorescence intensity. [ Resul] Lipid content of MⅡ stage was significantly lower than that in GV stage （66.63 ± 1. 197 vs. 70.81 ± 1. 328, P 〈 0.05 ）. Tbere was no significant dift）rence in lipid distribution pauems in oocytes of GV stage and MⅡ stage （ peripheral： 3. 148 ± 2. 298 vs. 5. 342 ± 3. 858 ; senti-peripheral： 30.85 ± 4. 042 vs. 35.70 ± 4. 265 ; 66.00 ± 3. 257 vs. 60.60 ± 3. 933, P 〉 0.05 ）. [ Conclusion] During the maturation process of the water buffalo oocytes from immature GV stage to mature MⅡ stage, the lipid content is significantly reduced, and the lipid distribution pattern is not changed before and after the vitro maturation of the oocytes.

关 键 词：水牛 GV期卵母细胞 MⅡ期卵母细胞 脂肪含量 脂肪分市 

分 类 号：S828.35[农业科学—畜牧学]