皮骨在斑马鱼侧线系统胚后发育中的作用  

The Role of Dermal Bone in Post-embryonic Development of Lateral Line in Zebrafish

作　　者：陈团结 段钦文 范纯新[1,2,3,4] Chen Tuanjie;Duan Qinwen;Fan Chunxin(Institute for Marine Biosystem and Neurosciences,International Center of Marine Study,Shanghai Ocean University,Shanghai,201306;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai,201306;Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources,Ministry of Education,Shanghai Ocean University,Shanghai,201306;Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture,Shanghai Ocean University,Shanghai,201306)

机构地区：[1]上海海洋大学,国际海洋研究中心,海洋生物系统和神经科学研究所,上海201306 [2]上海海洋大学,水产科学国家级实验教学示范中心,上海201306 [3]上海海洋大学,水产种质资源发掘与利用教育部重点实验室,上海201306 [4]上海海洋大学,农业部淡水水产种质资源重点实验室,上海201306

出　　处：《基因组学与应用生物学》2021年第1期153-158,共6页Genomics and Applied Biology

基　　金：国家自然科学基金项目(31772406);2019年度上海海洋大学国际海洋研究中心平台建设项目共同资助。

摘　　要：鱼类侧线系统的分布随其分类地位和不同的生存环境呈现多样性,这种多样性与侧线系统的胚后发育密切相关,但对于侧线系统胚后发育调控研究较少。本研究利用CRISPR/Cas9敲除斑马鱼(Brachydanio rerio var) edar基因,获得皮骨缺陷的突变体,进而利用染色技术分析鳞片和侧线管道在侧线胚后发育中的作用。利用茜素红对硬骨进行染色,我们发现edar突变体头部侧线管道不完整或缺失,躯干鳞片显著减少。经DASPEI对毛细胞进行染色,发现侧线管道不完整的头部管道神经丘变小,躯干鳞片缺失的表面神经丘无法形成缝合线。最后,利用qPCR对突变体WNT信号相关基因的表达进行分析,发现edar突变体中tcf7和lgr5表达显著下调。综上结果表明:斑马鱼的头部侧线管道和躯干的鳞片在侧线系统的胚后发育中起到重要的作用。The distribution of the lateral lineal system of fish varies with their taxonomic status and living environment. This diversity is closely related to the postembryonic development of the lateral lineal system, but there are few studies on the regulation of the postembryonic development of the lateral lineal system. In this study, the edar gene of zebrafish was knocked out by CRISPR/Cas9 to obtain skin and bone defect mutants, and then the role of scales and lateral line canals in lateral postembryonic development was analyzed by staining technique. Alizarin red was used to stain the bone. We found that edar mutants had incomplete or missing cranial lateral line canals and significantly reduced trunk scales. Hair cells were stained by DASPEI, and we found that the cranial neuromasts with incomplete lateral line canals became smaller, and the posterior lateral line superficial neuromasts of without trunk scales could not form stitch lines. Finally, qPCR was used to analyze the expression of WNT signaling related genes in the mutants. And it was found that tcf7 and lgr5 were down-regulated significantly in the edar mutants.In conclusion, the results showed that the cranial lateral line canals and the scales of the trunk of zebrafish play an important role in the postembryonic development of the lateral lineal system.

关 键 词：CRISPR/Cas9 edar突变体 侧线管道 缝合线 WNT信号 

分 类 号：S917.4[农业科学—水产科学]