斑马鱼卵母细胞的体外成熟及成熟卵的受精发育  

In Vitro Oocyte Maturation in the Zebrafish, Brachydanio rerio, and the Fertilization and Development of the Mature Egg

作　　者：李书鸿[1] 毛钟荣 韩文[1] 孙志远 闫维 陈惠萍[1] 严绍颐[1] 

机构地区：[1]中国科学院发育生物学研究所

出　　处：《生物工程学报》1993年第4期314-319,共6页Chinese Journal of Biotechnology

基　　金：国家“七.五”攻关;自然科学基金

摘　　要：采用体外培养的方法，研究斑马鱼卵母细胞的成熟过程。Ⅵ时相初级卵母细胞在０．５μｇ／ｍｌ １７ａ－羟基孕酮的ＥＭ－１９９培养液中，８０％氧气，２５℃的体外培养条件下，在４０ｍｉｎ内，胚泡逐渐由卵母细胞中央至动物极边缘１／２处移到动物边缘，进入Ⅴ时相卵母细胞。３０ｍｉｎ后胚泡破裂，胚泡破裂率为５９％，此种卵母细胞继续培养２ｈ才完全成熟，成熟卵不能不滤泡膜中自然排出。The in vitro maturation proccess of zebrafish oocyte was investigated. When in medium EM-199 containing 0.5μg/ml of 17α-hydroxyprogesterone, under the conditions of 80% O_2, 25℃, the germinal vesicles of the oocytes in stage Ⅳ migrated from midway between the center and the periphery to the periphery in 40 minutes and the oocytes went into stage V. 30min. later, the oocytes underwent germinal vesicle breakdown (GVBD)with a GVBD% of 59%. Two hours were needed for such oocytes to complete their final maturation. The mature eggs can not come off from the follicle layer surrounding them naturally(ovulation). By removing the follicle and adding active sperms for insemination, we could make the mature eggs fertilized. The chorion elevated and blastoderm formed on the animal pole. The cleavage and development of the fertilized eggs followed are just the same as that of naturally maturated fertilized eggs. Using the blastula as a successful fertilization of the in vitro maturated egg, the fertilization rate is 78%. This is the first report on the successful oocyte final maturation in vitro in zebrafisb. The establishment of oocyte in vitro maturation technique has made grounds for the further investigation of the transfer of foreign genes in the germinal vesicles of the oocytes.

关 键 词：卵母细胞 成熟 斑马鱼 受精 体外 

分 类 号：Q78[生物学—分子生物学]