The Function Study Of LncRNA Ihha-as In Early Embryonic Development Of Zebrafish

In recent years,with the development of bioinformatics and genomics,it has been found that not all transcription products have coding functions.Long non-coding RNA are RNA molecules with the length of more than 200 nt,which do not have encoding protein function,but it can regulate gene expression at multiple biological levels.ihha-as is a single-exon antisense long non-coding RNA that exists near the coding gene ihha.This paper revealed that ihha plays a very important role in the development and metabolism of zebrafish.However,the role of its antisense lnc RNA ihha-as in the early embryonic and cardiovascular development of zebrafish has not been reported.In light of this,the present research aims to determine the role of ihha-as in the early embryonic and cardiovascular development of zebrafish.In order to study the role of ihha-as in the early embryonic and cardiovascular development of zebrafish.We performed some research work as follows.First,bioinformatics analysis of ihha-as was carried out;secondly,q PCR and in situ hybridization were used to determine the Spatial and temporal expression patterns of ihha-as in zebrafish;then the effects of ihha-as overexpression on zebrafish's early development and cardiovascular marker genes expression were observed;finally,in situ hybridization and q PCR were used to detect morpholino interference,and ihha-as rescue in cardiovascular morphology,the expression of the sense coding gene ihha of ihha-as,and the expression of cardiovascular development marker genes.The results show that ihha-as is a highly conserved antisense lnc RNA and was located in zebrafish chromosome 9;the temporal and spatial expression shows that ihha-as has certain expression in different developmental stages of zebrafish and in different organs of mature fish,especially in the heart and brain;after overexpression,zebrafish embryos have no obvious morphological and functional changes,whereas the cardiovascular marker genes of amhc,vmhc,cmlc2 and flk1 are all up-regulated;after ihha-as-MO knockdown,zebrafish showed symptoms such as pericardial edema,heart hemorrhage and bone bending,and the expression of cardiovascular marker genes was significantly down-regulated.In situ hybridization showed that the heart was not circularized and cardiovascular development slowed down after knockdown;after ihha-as rescued,the zebrafish malformation symptoms were alleviated,the expression of cardiovascular marker genes increased,and the abnormal cardiovascular development was improved;after the final knockdown of ihha-as,the expression of the coding gene ihha increased significantly,and the expression decreased significantly after salvage.These results indicated that the long non-coding RNA ihha-as regulates the early cardiovascular development of zebrafish by affecting the expression of ihha.This research provides a certain experimental evidence for the molecular mechanism and biological functions of lnc RNA in the early cardiovascular development of zebrafish.