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细胞周期蛋白A2在雌核发育银鲫和两性生殖彩鲫的卵母细胞成熟过程中差异表达。

Cyclin A2 is differentially expressed during oocyte maturation between gynogenetic silver crucian carp and gonochoristic color crucian carp.

作者信息

Xie Jing, Wen Jian-Jun, Yang Zhong-An, Wang Hong-Ying, Gui Jian-Fang

机构信息

State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.

出版信息

J Exp Zool A Comp Exp Biol. 2003 Jan 1;295(1):1-16. doi: 10.1002/jez.a.10209.

Abstract

Silver crucian carp (Carassius auratus gibelio) is a unique gynogenetic fish. Because of its specific genetic background and reproduction mode, it is an intriguing model system for understanding regulatory mechanism of oocyte maturation division. It keeps its chromosomal integrity by inhibiting the first meiotic division (no extrusion of the first pole body). The spindle behavior during oocyte maturation is significantly different from that in gonochoristic fish. The chromosomes are first arranged in a tripolar spindle, and then they turn around and are reunited mutually to form a normal bipolar spindle. A new member of the fish A-type cyclin gene, cyclin A2, has been isolated by suppression of subtractive hybridization on the basis of its differential transcription in fully-grown oocytes between the gynogenetic silver crucian carp and gonochoristic color crucian carp. There are 18 differing amino acids in the total 428 residues of cyclin A2 between the two forms of crucian carps. In addition, cDNAs of cyclin Al and cyclin B have also been cloned from them. Thus two members of A-type cyclins, cyclin Al and cyclin A2, are demonstrated to exist in fish, just as in frog, humans, and mouse. Northern blotting reveals that cyclin A2 mRNA is more than 20-fold and cyclin A1 mRNA is about 2-fold in fully grown oocytes of gynogenetic silver crucian carp compared to gonochoristic color crucian carp. However, cyclin B does not show such a difference between them. Western blot analysis also shows that the cyclin A2 protein stockpiled in fully grown oocytes of gynogenetic crucian carp is much more abundant than in gonochoristic crucian carp. Moreover, two different cyclin A2 expression patterns during oocyte maturation have been revealed in the two closely related crucian carps. For color crucian carp, cyclin A2 protein is translated only after hormone stimulation. For silver crucian carp, cyclin A2 protein can be detected throughout the process of maturation division. The different expression of cyclin A2 may be a clue to understanding the special maturation division of gynogenetic silver crucian carp.

摘要

银鲫(Carassius auratus gibelio)是一种独特的雌核发育鱼类。由于其特定的遗传背景和繁殖方式,它是理解卵母细胞成熟分裂调控机制的一个有趣的模型系统。它通过抑制第一次减数分裂(第一极体不排出)来保持其染色体完整性。卵母细胞成熟过程中的纺锤体行为与雌雄异体鱼类显著不同。染色体首先排列成三极纺锤体,然后它们扭转并相互重新结合形成正常的双极纺锤体。鱼类A型细胞周期蛋白基因的一个新成员,细胞周期蛋白A2,已通过抑制性消减杂交法分离出来,其依据是雌核发育银鲫和雌雄异体彩鲫的完全成熟卵母细胞中的差异转录。两种鲫鱼的细胞周期蛋白A2在总共428个氨基酸残基中有18个不同的氨基酸。此外,还从它们中克隆了细胞周期蛋白A1和细胞周期蛋白B的cDNA。因此,正如在青蛙、人类和小鼠中一样,证明鱼类中存在A型细胞周期蛋白的两个成员,即细胞周期蛋白A1和细胞周期蛋白A2。Northern印迹分析表明,与雌雄异体彩鲫相比,雌核发育银鲫完全成熟卵母细胞中的细胞周期蛋白A2 mRNA含量高出20多倍,细胞周期蛋白A1 mRNA含量约高出2倍。然而,细胞周期蛋白B在它们之间没有显示出这种差异。蛋白质免疫印迹分析还表明,雌核发育鲫鱼完全成熟卵母细胞中储存的细胞周期蛋白A2蛋白比雌雄异体鲫鱼中的丰富得多。此外,在两种亲缘关系密切的鲫鱼中揭示了卵母细胞成熟过程中细胞周期蛋白A2的两种不同表达模式。对于彩鲫,细胞周期蛋白A2蛋白仅在激素刺激后才翻译。对于银鲫,在成熟分裂的整个过程中都可以检测到细胞周期蛋白A2蛋白。细胞周期蛋白A2的不同表达可能是理解雌核发育银鲫特殊成熟分裂的一条线索。

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