Abstract
Long noncoding RNAs (lncRNAs) have been shown to play crucial roles in various life processes, including circadian rhythms. Although next generation sequencing technologies have facilitated faster profiling of lncRNAs, the resulting datasets require sophisticated computational analyses. In particular, the regulatory roles of lncRNAs in circadian clocks are far from being completely understood. In this study, we conducted RNA-seq-based transcriptome analysis of zebrafish larvae under both constant darkness (DD) and constant light (LL) conditions in a circadian manner, employing state-of-the-art computational approaches to identify approximately 3220 lncRNAs from zebrafish larvae, and then uncovered 269 and 309 lncRNAs displaying circadian rhythmicity under DD and LL conditions, respectively, with 30 of them are coexpressed under both DD and LL conditions. Subsequently, GO, COG, and KEGG pathway enrichment analyses of all these circadianly expressed lncRNAs suggested their potential involvement in numerous biological processes. Comparison of these circadianly expressed zebrafish larval lncRNAs, with rhythmically expressed lncRNAs in the zebrafish pineal gland and zebrafish testis, revealed that nine (DD) and twelve (LL) larval lncRNAs are coexpressed in the zebrafish pineal gland and testis, respectively. Intriguingly, among peptides encoded by these coexpressing circadianly expressed lncRNAs, three peptides (DD) and one peptide (LL) were found to have the known domains from the Protein Data Bank. Further, the conservation analysis of these circadianly expressed zebrafish larval lncRNAs with human and mouse genomes uncovered one lncRNA and four lncRNAs shared by all three species under DD and LL conditions, respectively. We also investigated the conserved lncRNA-encoded peptides and found one peptide under DD condition conserved in these three species and computationally predicted its 3D structure and functions. Our study reveals that hundreds of lncRNAs from zebrafish larvae exhibit circadian rhythmicity and should help set the stage for their further functional studies.
Highlights
The circadian clock, an endogenous time-keeping mechanism, regulates unique 24-h rhythms of metabolism, physiology and behavior [1]
Some genes peak during the morning, some during the evening, and others at night [46,47,48]. We investigated whether these zebrafish larval Long noncoding RNAs (lncRNAs) exhibit peak expression during morning, evening, and night
The correlative expression profiles of these 269 circadianly expressed larval lncRNAs are shown in a heat map (Figure 1A), while their phases are exhibited as BioDare2 plots (Figure 1B)
Summary
The circadian clock, an endogenous time-keeping mechanism, regulates unique 24-h rhythms of metabolism, physiology and behavior [1]. The suprachiasmatic nucleus (SCN) of the hypothalamus hosts the master clock that drives the circadian rhythms in various tissues and organs [2]. A variety of model organisms including the fruit fly (Drosophila melanogaster) and the zebrafish (Danio rerio) [4] have been used to study the operating mechanisms of circadian clocks. The fruit fly is an ideal organism to investigate circadian clocks in insects [5] because of its easy genetic manipulation, breeding in a controlled environment, and monitoring of locomotor activities [6]. The zebrafish is an attractive organism to study the circadian clock in vertebrates [7,8,9]. Zebrafish embryos/larvae allow for studying circadian rhythms independent of feeding
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