水生生物学报2017，Vol.41Issue(5)：984-991,8.DOI:10.7541/2017.123
银鲫卵母细胞体外诱导成熟技术的建立
DEVELOPMENT OF IN VITRO MATURATION TECHNOLOGY FOR GIBEL CARP OOCYTES
摘要
Abstract
In vitro oocytes maturation is a key and efficient technology for gene and cellular micromanipulation in oo-cytes of fish. In this study, we used gibel carp oocytes as studying materials to develop an efficient in vitro induction maturation technology. Based on previously established germinal vesicle (GV) isolation and premeiotic chromosome preparation methods , four different stage oocytes including GV0, GV1, GV2 and GV3 were firstly distinguished by GV migration and premeiotic chromosome condensation status, and the maturation rate (％), cleavage rate (％) and hatching rate (％) were analyzed. GV1, GV2 and GV3 oocytes had the capacity for in vitro induction maturation, and GV1 oocytes were the optimal stage oocytes because they have longer time for micromanipulation. We found that the optimal induction conditions for GV1 oocytes in vitro were 12 hours at 23℃ in pH 8.5 Gey's balanced salt solution(GBSS) with 1 μg/ml DHP (17α, 20β-dihydroxy-4-pregne-3-one), by which up to 55.5％ hatching rate was reached from these stripped follicle membrane oocytes. Moreover, we injected the GFP-zfh2af1o mRNA into the GV1 oocytes, and found that the whole process of in vitro maturation, fertilization and embryo development of the micro-manipulated GV1 oocytes could be tracked by the expression of GFP, and normal fertilization and embryo develop-ment could be produced from the induced mature eggs. Therefore, we established an efficient platform for studying oo-cyte development and gene and cellular micromanipulation in this fish.关键词
银鲫/卵母细胞成熟/体外诱导/生发泡/显微注射Key words
Gibel carp/Oocyte/In vitro maturation/Germinal vesicle/Micromanipulation分类
生物科学引用本文复制引用
李志,汪洋,周莉,李熙银,张晓娟,周剑,桂建芳..银鲫卵母细胞体外诱导成熟技术的建立[J].水生生物学报,2017,41(5):984-991,8.基金项目
中国科学院战略性先导科技专项 (XDA08030201)资助 Supported by the Strategic Priority Research Program of Sciences (XDA08030201) （XDA08030201）
