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《Fudan University Journal of Medical Sciences》 2008-03
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The effects of hand2 knock-down on the zebrafish embryonic development

SUN Shu-na1,GUI Yong-hao1,SONG Hou-yan2,JIANG Qiu2,WANG Yue-xiang2(1Department of Cardiology,Children's Hospital,Fudan University,Shanghai 200032,China;2Key Laboratory of Molecular Medicine,Ministry of Education-Department of Molecular Genetics,Shanghai Medical College,Fudan University,Shanghai 200032,China)  
Objective To detect the hand2 expressing pattern in zebrafish embryos,and to demonstrate that microinjecting morpholino oligonucleotides(MO)would be an effective method to block the hand2 expression and observe the abnormal phenotypes induced by hand2 knock-down.The present study is expected to provide the clue for the functional study on hand2.Methods Whole-mount in situ hybridization was performed to detect the expression pattern of hand2.Zebrafish fertilized eggs were divided into four groups(Con MO microinjected group,hand2 MO microinjected group,hand2-GFP mRNA microinjected group and hand2-GFP mRNA+hand2 MO microinjected group).The GFP expression of hand2-GFP mRNA microinjected group and hand2-GFP mRNA+hand2 MO microinjected group were detected by fluorescent microscope to evaluate the efficiency of hand2 knock-down.The development of Con MO microinjected embryos and hand2 MO microinjected embryos were observed under microscope.Results The hand2 expression were observed in lateral mesoderm,pharyngeal arches,heart and fins.Strong GFP expression was observed in hand2-GFP mRNA microinjected group at 8 hpf(hours post fertilization).Compared with hand2-GFP mRNA microinjected group,the expression of GFP in hand2-GFP mRNA+hand2 MO microinjected group was obviously reduced.Compared with Con MO group,the hand2 MO injected group showed apparent malformation at the heart,the vessel,the blood cells,the fin and the somite.Conclusions Microinjecting hand2 MO can effectively knock-down hand2.hand2 plays an important role in the development of the heart,the vessel,the blood cell,the fin and the somite in zebrafish.
【Fund】: 上海市科委专项课题“先心病产前诊断技术的多中心研究和评价”项目(054119512)
【CateGory Index】: Q344
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