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

作　　者：金璐[1,2] 丁可军[2] 孙琮杰 陈露 周艳华[1] 舒莉萍[1,3,4] 何志旭 JIN Lu;DING Kejun;SUN Congjie;CHEN Lu;ZHOU Yanhua;SHU Liping;HE Zhixu(Tissue Engineering and Stem Cell Researeh Center,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Pediatrics,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Immunology,Basic Medical College,Guizhou Medical University,Guiyang 550004,Guizhou,China;Laboratory Animal Center,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学组织工程和干细胞实验中心,贵州贵阳550004 [2]贵州医科大学儿童医学中心,贵州贵阳550004 [3]贵州医科大学基础医学院免疫学教研室,贵州贵阳550004 [4]贵州医科大学实验动物中心,贵州贵阳550004

出　　处：《贵州医科大学学报》2018年第10期1178-1182,1191,共6页Journal of Guizhou Medical University

基　　金：国家自然科学基金资助项目(31360285)

摘　　要：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 thine 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 se- lected 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 3.7hpf and 9 -24 hpf, and so did in protonephridia in 18 -30 hpf. There was no expression after 36hpf. 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 in 18 -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：斑马鱼 HoxB4基因 FLASH基因 转基因 原位杂交 

分 类 号：Q75[生物学—分子生物学]