English

Abstract

Long non-coding RNAs (lncRNA) are increasingly being regard as potential key regulators of biological process, however, little is known about the function of most of them. The involvement of a novel lncRNA (ENSDART000000150571), previously named TCONS_00028652, in intersegmental vessel development was investigated in this study. TCONS_00028652, having a single exon (568 bp), is located in chromosome 16 at position from 15786804 to 15786237, which was previously identified as an embryonic and adult heart-enrichedlncRNA. Bioinformatics analysis and annotation of TCONS_00028652 was performed using online databases. Its protein-coding potential was assessed using online softwares: the Coding Potential Assessing Tool (CPAT) and the Coding Potential Calculator (CPC). To verify its real existence, a cloned fragment was proliferated by designing primers against 5’ and 3’ exon-flanking sequence. Subsequently, spatiotemporal expression during zebrafish embryonic development was determined using real-time quantitative PCR (qPCR) and whole mount in situ hybridization. We found that lncRNA TCONS_00028652 was generally expressed throughout early stages of zebrafish embryonic development and predominantly in embryonic brain, tail, and heart. Knockdown of TCONS_00028652 using morpholino oligonucleotides (MO) resulted in intersegmental vessel defects, suggesting that TCONS_00028652 is indispensable for embryonic vascular development in zebrafish. Using Tg (flil:EGFP) transgenic fish expressing a cardiovascular marker gene, loss of function experiments confirmed that TCONS_00028652 was involved in embryonic intersegmental vessel development. Our results may lead to valuable understanding of lncRNAs functions in zebrafish embryonic development and molecular mechanisms of embryonic cardiovascular development. © 2021 Friends Science Publishers