摘要
为探究骨形态发生蛋白15(Bone morphogenetic protein 15, bmp15)在银鲫(Carassius gibelio Bloch) F系卵巢发育过程中的表达特征,研究采用cDNA末端快速扩增(RACE)方法首先从银鲫F系卵巢cDNA中克隆了2个bmp15部分同源基因(Homeologs),将其命名为Cgbmp15a和Cgbmp15b,它们各自具有3个不同等位基因。不同脊椎动物Bmp15蛋白序列多重比对和系统进化树均表明,银鲫为异源六倍体,在其进化历程中发生了两轮多倍化,其中早期的异源多倍化整合了来自2个原始祖先的染色体组,导致银鲫和鲫的四倍体共同原始祖先基因组中包含bmp15a和bmp15b;随后第二轮同源多倍化最终导致银鲫存在6个bmp15等位基因。bmp15基因及其邻近基因同线性分析表明,银鲫在形成异源多倍体后,其基因组发生了复杂的变化,包括bmp15邻近基因的丢失。Cgbmp15a和Cgbmp15b均主要在卵巢中表达,在皮质泡期卵母细胞中表达量最高;无论是在银鲫成体组织还是不同发育阶段的卵子中, Cgbmp15a的表达水平显著高于Cgbmp15b的表达水平。在孕酮激素DHP体外诱导银鲫F系GV1期卵母细胞6h后, Cgbmp15a开始上调表达,诱导8h后达到最高表达水平,随后下降;而Cgbmp15b在诱导8h后表达水平有微弱上调,随后下降。上述的结果表明, Cgbmp15a和Cgbmp15b在银鲫F系成体组织和不同发育和成熟阶段的卵子中,均表现出偏性表达的特征,暗示Cgbmp15a在银鲫的卵母细胞发育和成熟中起着主要作用。
Bone morphogenetic protein 15(bmp15) plays key roles in regulating development and function maintenance of reproductive system in vertebrates. In this study, we first identified two divergent bmp15 homeologs from gibel carp(Carassius gibelio Bloch) clone F. Both Cgbmp15 a and Cgbmp15 b possess 3 different alleles, which were obtained from ovary cDNA by using rapid-amplification of cDNA ends(RACE). Multiple sequence alignment of Bmp15 protein in vertebrates and phylogenetic analysis of gibel carp, crucian carp, and zebrafish bmp15 genomic sequences indicated that two rounds of polyploidization had been occurred during allo-hexaploid gibel carp evolution. The early polyploidy produced the two divergent bmp15 a and bmp15 b, and the late polyploidy generated the 6 Cgbmp15 alleles of gibel carp. Synteny analysis of genomic regions around bmp15 gene in vertebrates suggested that the complex genomic changes might occur after the polyploidization in gibel carp, such as the loss of bmp15 adjacent genes.Cgbmp15 a and Cgbmp15 b were predominantly expressed in ovary with the highest level in cortical alveolus oocytes.The Cgbmp15 a level is remarkably higher than Cgbmp15 b in adult tissues and oocytes at different developmental stages(P<0.05). Moreover, Cgbmp15 a increased at 6 hours and reached peak level at 8 hours after DHP induction in vitro, while Cgbmp15 b slightly increased at 8 hours after DHP induction. These results indicate that Cgbmp15 a play a major role in gibel carp oocyte development and maturation.
作者
江诗雨
汪洋
周莉
陈帆
李志
桂建芳
JIANG Shi-Yu;WANG Yang;ZHOU Li;CHEN Fan;LI Zhi;GUI Jian-Fang(State Key Laboratory of Freshwater Ecology and Biotechnology,the Innovative Academy of Seed Design,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,China;University of Chinese Academy of Sciences,Beijing 100049,China;University of Chinese Academy of Sciences Beijing 100049,China)
出处
《水生生物学报》
CAS
CSCD
北大核心
2020年第3期518-527,共10页
Acta Hydrobiologica Sinica
基金
中国科学院战略性先导科技专项(XDA08030201)
现代农业产业技术体系(CARS-45-07)资助。
