CAJ | 학술논문

为研究转化生长因子β(Transforming growth factor β, 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。
위연구전화생장인자β(Transforming growth factor β, TGFβ)1대반마어배태발육적조공작용,통과NCBI획득TGF-β1기인서렬, TGF-β1 cDNA전장1571 bp,편마377개안기산。계통진화수분석발현, TGF-β단백안조불동적류형엄격취류,반마어TGF-β1여기타어류적TGF-β1취집도일개분지,재진화중비상보수。대반마어배태진행RT-PCR화Real-Time PCR검측현시, TGF-β1기인위모원표체기인,재분절기지전적표체수평비교저,이종인낭기개시지속고수평적표체。배태정체원위잡교발현, TGF-β1기인재반마어24 hpf 배태중개시유특이신호출현, TGF-β1기인적표체주요분포재시궁、측선원기、이낭、후각기판、심장화전신등처,표명TGF-β1기인가능삼여반마어배태면역조절、순배계통발육화측선형성。용저양처리반마어배태,발현저양처리24h후반마어배태발육연지。이용Real-Time PCR화배태정체원위잡교검측발현,저양처리후발육연지적반마어배태중TGF-β1 mRNA표체량교상양조현저강저。이상결과표명, TGF-β1기인삼여반마어배태발육조공,병차가능여저양처리후반마어배태발육연지유관。연구결과장위심입연구반마어TGF-β1기인적공능전정기출。
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.