lncRNA ihha-as在斑马鱼早期胚胎发育中的功能研究  

The Function of lncRNA Ihha-as in Early Zebrafish Embryonic Development

作　　者：欧阳奇 李小灯 远长悦 李婧琳 韩梅 胡翔 OUYANG Qi;LI Xiao-deng;YUAN Chang-yue;LI Jing-lin;HAN Mei;HU Xiang(State Key Laboratory of Developmental Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Changsha 410081, China)

机构地区：[1]湖南师范大学生命科学学院,省部共建淡水鱼类发育生物学国家重点实验室,中国长沙410081

出　　处：《湖南师范大学自然科学学报》2021年第6期85-91,共7页Journal of Natural Science of Hunan Normal University

基　　金：长沙市自然科学基金(kq2007013);省部共建淡水鱼类发育生物学国家重点实验室开放基金(2020KF003);湖南师范大学大学生创新创业基金(2021118)。

摘　　要：为了探究lncRNA ihha-as在斑马鱼早期胚胎和心脏发育中的作用,利用NCBI blast和UCSC Genome Browser对ihha-as进行序列保守性分析,通过半定量PCR和qPCR及原位杂交实验确定ihha-as在斑马鱼中的时空表达,运用ihha-as-morpholino敲低和ihha-as挽救实验探究ihha-as的生物学功能。研究发现ihha-as为斑马鱼9号染色体产生的一个具有极高保守性的反义lncRNA且在斑马鱼心脏中有着特异性表达;ihha-as-morpholino敲低后发现斑马鱼出现心包水肿、心脏静脉窦出血和骨骼弯曲等症状,进一步通过qPCR检测心脏表达的相关基因,发现有所下调;利用斑马鱼胚胎原位杂交证实,敲低后心脏发育变慢、心脏未环化且形态不完整;显微共注射ihha-as进行挽救后,斑马鱼畸形症状有所缓解,心脏发育异常得到改善。研究结果表明lncRNA ihha-as对斑马鱼心脏系统发育至关重要。In order to explore the role of lncRNA ihha-as in the early embryonic and heart development of zebrafish,the NCBI blast and UCSC Genome Browser were used to analyze the conservation of ihha-as sequence,and the temporal and spatial expression of ihha-as in the zebrafish were detected through the semi-quantitative PCR,qPCR and in situ hybridization experiments.The ihha-as-morpholino knockdown experiment and the ihha-as rescue experiment were performed to explore the ihha-as biological functions.This research has demonstrated that the ihha-as is a highly conserved antisense lncRNA produced by the zebrafish chromosome 9,which is specifically expressed in the heart of zebrafish.After the ihha-as-morpholino was knocked down,it was found that the zebrafish had symptoms of pericardial edema,cardiac sinus bleeding and skeletal curvature.The qPCR was further used to detect the related genes expressed in the heart,and it was found that the genes were regulated down.It was confirmed by the in situ hybridization of zebrafish embryos that after the ihha-as-morpholino knockdown experiment,the heart development slowed down,the morphology of heart was incomplete and the heart was not circularized.After the rescue of ihha-as microinjection,the zebrafish malformation symptoms were relieved and the cardiac dysplasia was improved.The results indicate that the lncRNA ihha-as is essential for the development of the zebrafish heart system.

关 键 词：lncRNA ihha-as 斑马鱼 心脏发育 MORPHOLINO 

分 类 号：S917.4[农业科学—水产科学]