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

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

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

Abstract:

Keywords:

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

DOI: 10.19367/j.cnki.1000-2707.2018.10.012

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