贵州医科大学学报

2018, v.43;No.217(10) 1178-1182+1191

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FLASH基因在野生型斑马鱼早期表达以及与HoxB4基因的关系
The Early Expression of FLASH Gene in Wild Zebrafish and the Relationship with HoxB4 Gene

金璐;丁可军;孙琮杰;陈露;周艳华;舒莉萍;何志旭;
JIN Lu;DING Kejun;SUN Congjie;CHEN Lu;ZHOU Yanhua;SHU Liping;HE Zhixu;Tissue Engineering and Stem Cell Research Center,Guizhou Medical University;Department of Pediatrics,Guizhou Medical University;Department of Immunology,Basic Medical College,Guizhou Medical University;Laboratory Animal Center,Guizhou Medical University;

摘要(Abstract):

目的:观测FLASH基因在Tuebingen野生型斑马鱼胚胎早期的表达及与Hox B4基因的关系。方法:以Hox B4转基因斑马鱼系为研究对象,分为实验组、实验对照组和空白对照组,实验组为Hox B4转基因斑马鱼系与Cre鱼系交配所得的过表达Hox B4的带有EGFP荧光的胚胎(Tg:Lmo2-Cre,Lmo2-LDEL-Hoxb4-EGFP),实验对照组为EGFP转基因斑马鱼系与Cre鱼系交配所得的表达EGFP荧光的胚胎(Tg:Lmo2-Cre,Lmo2-LDEL-EGFP),空白对照组为Tuebingen野生型斑马鱼的胚胎;采用FLASH反义mRNA探针做原位杂交并观察其表达。结果:观察到Tuebingen组中3. 7、9~24 hpf的神经外侧板、头部和脊索及18~30 hpf的原肾管可见黑色的阳性杂交信号,36 hpf后未见; 18~72 hpf的空白对照组与实验对照组间FLASH表达水平无差异,实验组18~30 hpf神经系统、造血系统FLASH的表达下调。结论:FLASH基因可能是Hox B4基因的下游靶基因。
Objective: To study the expression of FLASH gene in the early stage of embryos of Tuebingen wild zebrafish and its relationship with HoxB4 gene. Methods: HoxB4 transgenic zebrafish lines were divided into three groups. The Tg:(Lmo2-Cre; Lmo2-LDEL-Hoxb4-EGFP) zebrafish were selected as experimental group; the Tg:(Lmo2-Cre; Lmo2-LDEL-EGFP) transgenic zebrafish were selected as experimental control group; the Tubingen wild type zebrafish were selected as blank control group. FLASH antisense mRNA probe was used for hybridization in situ and its expression was observed. Results: For the Tubingen wild type zebrafish,the black positive hybridization signal was observed in nerve lateral plate,head and sagittal in 3. 7 hpf and 9 ~ 24 hpf,and so did in protonephridia in 18 ~ 30 hpf. There was no expression after 36 hpf. There was no significant difference between the blank control group and the experimental control group in 18 ~ 72 hpf. And in the experimental group,the expression of FLASH gene in the nervous system and hematopoietic system was down-regulated in18 ~ 30 hpf. Conclusion: The FLASH gene may be the downstream target gene of HoxB4 gene and it is helpful to understand the expression of FLASH gene in the early stage of wild-type zebrafish and lay a foundation for the study of FLASH gene-related functions.

关键词(KeyWords): 斑马鱼;HoxB4基因;FLASH基因;转基因;原位杂交
zebrafish;HoxB4 gene;FLASH gene;transgenosis;hybridization in situ

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基金项目(Foundation): 国家自然科学基金资助项目(31360285)

作者(Author): 金璐;丁可军;孙琮杰;陈露;周艳华;舒莉萍;何志旭;
JIN Lu;DING Kejun;SUN Congjie;CHEN Lu;ZHOU Yanhua;SHU Liping;HE Zhixu;Tissue Engineering and Stem Cell Research Center,Guizhou Medical University;Department of Pediatrics,Guizhou Medical University;Department of Immunology,Basic Medical College,Guizhou Medical University;Laboratory Animal Center,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2018.10.012

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