摘要
为研究转化生长因子β(Transforming growth factorβ,TGFβ)1对斑马鱼胚胎发育的调控作用,通过NCBI获得TGF-β1基因序列,TGF-β1 cDNA全长1571 bp,编码377个氨基酸。系统进化树分析发现,TGF-β蛋白按照不同的类型严格聚类,斑马鱼TGF-β1与其他鱼类的TGF-β1聚集到一个分支,在进化中非常保守。对斑马鱼胚胎进行RT-PCR和Real-Time PCR检测显示,TGF-β1基因为母源表达基因,在分节期之前的表达水平比较低,而从咽囊期开始持续高水平的表达。胚胎整体原位杂交发现,TGF-β1基因在斑马鱼24 hpf胚胎中开始有特异信号出现,TGF-β1基因的表达主要分布在腮弓、侧线原基、耳囊、嗅觉基板、心脏和前肾等处,表明TGF-β1基因可能参与斑马鱼胚胎免疫调节、循环系统发育和侧线形成。用低氧处理斑马鱼胚胎,发现低氧处理24h后斑马鱼胚胎发育延迟。利用Real-Time PCR和胚胎整体原位杂交检测发现,低氧处理后发育延迟的斑马鱼胚胎中TGF-β1 mRNA表达量较常氧组显著降低。以上结果表明,TGF-β1基因参与斑马鱼胚胎发育调控,并且可能与低氧处理后斑马鱼胚胎发育延迟有关。研究结果将为深入研究斑马鱼TGF-β1基因的功能奠定基础。
Transforming growth factor-β1 (TGF-β1) is a pleiotropic cytokine and is involved in the regulation of key events in development, disease, and tissue repair. Here we explored the expression of TGF-β1 in zebrafish of which the full cDNA consists of 1571 bp and encodes 377aa. The sequence of this gene was obtained from NCBI. Phylogenetic analysis revealed that TGF-βs were classified strictly with the same type of proteins. TGF-β1 of zebrafish was classified on the same branch of the phylogenetic tree with TGF-β1s of other fish. RT-PCR of this gene at different embryonic stages showed that TGF-β1 existed in unfertilized eggs of zebrafish. The expression started to increase at 24hpf and maintained a high level until 72hpf. Whole-mount in situ hybridization (WISH) analysis showed that TGF-β1 transcripts could be first detected in embryos at 24hpf and increased noticeably at later developmental stages. TGF-β1 transcripts were mainly expressed in gill arches, lateral line primordium, otic vesicle, olfactory placode, heart and pronephros. These indicated that TGF-β1 might play an essential role in immunoregulation, lateral line formation and vasculogenesis. Furthermore, the effect of hypoxia on the expression of TGF-β1 in zebrafish embryogenesis was evaluated with RT-PCR, Real-Time PCR and WISH. After the hypoxia treatment the development of embryos was delayed and the expression of TGF-β1 in embryos decreased dramatically. These results suggested that TGF-β1 played an important role in zebrafish embryogenesis and might be related to the hypoxia-induced delay in zebrafish embryonic developement.
出处
《水生生物学报》
CAS
CSCD
北大核心
2014年第6期1054-1061,共8页
Acta Hydrobiologica Sinica
基金
国家自然科学基金项目(30972254,31272645)
中央高校基本科研业务费专项资金科研项目(CCNU12A02007)资助
