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Long non-coding RNA (lncRNA) is a novel emerging type of competitive endogenous RNA (ceRNA) that performs key functions in multiple biological processes. However, little is known about the roles of lncRNA under hypoxia stress in fish. Here, vascular endothelial growth factor-Aa (vegfaa) was cloned in rainbow trout (Oncorhynchus mykiss), with the complete cDNA sequence of 2914 bp, encoding 218 amino acids. The molecular weight of the protein was approximately 25.33 kDa, and contained PDGF and VEGF_C domains. Time-course and spatial expression patterns revealed that LOC110520012 was a key regulator of rainbow trout in response to hypoxia stress, and LOC110520012, miR-206-y and vegfaa exhibited a ceRNA regulatory relationship in liver, gill, muscle and rainbow trout liver cells treated with acute hypoxia. Subsequently, the targeting relationship of LOC110520012 and vegfaa with miR-206-y was confirmed by dual-luciferase reporter analysis, and overexpression of LOC110520012 mediated the inhibition of miR-206-y expression in rainbow trout liver cells, while the opposite results were obtained after LOC110520012 silencing with siRNA. We also proved that vegfaa was a target of miR-206-y in vitro and in vivo , and the vegfaa expression and anti-proliferative effect on rainbow trout liver cells regulated by miR-206-y mimics could be reversed by LOC110520012. These results suggested that LOC110520012 can positively regulate vegfaa expression by sponging miR-206-y under hypoxia stress in rainbow trout, which facilitate in-depth understanding of the molecular mechanisms of fish adaptation and tolerance to hypoxia. • The full-length cDNA sequence of vegfaa was 2914 bp encoding 218 amino acids. • LOC110520012 and vegfaa were highly expressed under hypoxia stress. • vegfaa was shown to a direct target of miR-206-y in vitro and in vivo. • LOC110520012 acted as a ceRNA of vegfaa by sponging miR-206-y. • The inhibitory effect of miR-206-y on rainbow trout liver cell proliferation could be alleviated by LOC110520012. [ABSTRACT FROM AUTHOR]

Hypoxia, largely triggered by global warming and water contamination, has become an environmental issue of great concern, posing a great threat to aquatic ecosystem. As one of the world's most economically important fish, rainbow trout (Oncorhynchus mykiss) is extremely intolerant of hypoxic environments, however, little is known about the roles of non-coding RNAs (ncRNAs) in the response of rainbow trout to hypoxia stress. Herein, effects of moderate (Tm12L) and severe hypoxia for 12 h (Ts12L) and 12 h reoxygenation on histology, biochemical parameters (antioxidant, metabolism and immunity) and transcriptome (lncRNA, miRNA and mRNA) in rainbow trout liver were investigated. We further validated the regulatory relationships between LOC110519952, novel-m0023-5p and glut1a via dual‑luciferase reporter, overexpression and silencing assays. Compared with Tm12L, the liver in Ts12L showed more severe oxidative damage. Anaerobic, lipid and protein metabolism was enhanced under hypoxia stress, especially in Ts12L. We also found that Tm12L could strengthen innate immune response, which was inhibited in Ts12L. Besides, several hypoxia-related genes (glut1a , vegfaa , hmox , epoa , foxo1a and igfbp1) and ceRNA networks were identified from 1824, 427 and 545 differentially expressed mRNAs, miRNAs and lncRNAs, including LOC118965299-novel-m0179-3p- epoa , LOC110519952-novel-m0023-5p- glut1a , MSTRG.7382.2-miR-184-y- hmox and LOC110520012-miR-206-y- vegfaa. Through in vitro and in vivo functional analysis, we demonstrated that glut1a is a target of novel-m0023-5p, and LOC110519952 can positively regulate glut1a by targeting novel-m0023-5p. Introduction of LOC110519952 could attenuate the promoting effects of novel-m0023-5p on rainbow trout liver cell viability and proliferation. This study highlights the differences in the regulatory mechanism of rainbow trout under different concentrations of hypoxia stress and provides valuable data for further research on the molecular mechanisms of fish adaptation to hypoxic environments. [Display omitted] • Anaerobic metabolism was strongly enhanced under severe hypoxia. • Innate immune response was inhibited under severe hypoxia. • Numerous hypoxia-related genes and lncRNA-miRNA-mRNA networks were identified. • LOC110519952 functioned as ceRNA to target glut1a by sponging novel-m0023-5p. • Overexpressed novel-m0023-5p promoted cell viability and proliferation. [ABSTRACT FROM AUTHOR]

Rainbow trout (Oncorhynchus mykiss) are a cold-water salmonid species that is highly susceptible to heat stress. Summer temperature stress is a common issue in trout aquaculture. To better understand the molecular mechanisms of the heat-stress response in the trout, we used label-free quantitative proteome techniques to identify differentially expressed proteins in the livers of rainbow trout exposed to heat stress. We identified 3362 proteins and 152 differentially expressed proteins (p < 0.05; fold-change >2). Of these, 37 were uniquely expressed in the heat-stress group and 35 were uniquely expressed in the control group. In addition, 42 proteins were significantly upregulated (fold-change >2) and 38 proteins were significantly downregulated (fold-change >2). GO (Gene Ontology) analysis indicated that these differentially expressed proteins were primarily expressed in the nucleus, extracellular matrix, and cytoplasm, and were associated with a variety of functions, including protein binding/bridging and enzyme facilitation. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis of the differentially expressed proteins showed that, during high temperature stress, many biological processes were extensively altered, particularly the estrogen signaling pathway, the complement and coagulation cascades, and the platelet activation pathway. Our study focused on the identification of a systematic approach for the characterization of regulatory networks. Our results provide a framework for further studies of the heat-stress response in fish. Unlabelled Image [ABSTRACT FROM AUTHOR]

Rainbow trout (Oncorhynchus mykiss) is an important economic fish in China. Skin color affects the economic value of trout. However, the molecular mechanism of the skin color variation between wild-type (WR) and yellow mutant rainbow trout (YR) is unclear. We sequenced mRNAs and miRNAs of dorsal skin to identify key color variation-associated mRNAs and miRNAs between WR and YR. Overall, 2060 out of 3625 differentially expressed genes were upregulated in YR, and 196 out of 275 differentially expressed miRNAs were downregulated in WR. We identified three key YR-upregulated genes related to the formation of xanthophores (GCH1 , SLC2A11 , and SOX10). Interestingly, several genes related to melanogenesis (TYR , TYRP1 , TYRP2 , MC1R , MITF , PMEL , SLC45A2 , and OCA2) were downregulated in WR. Integrated analysis identified five miRNAs that target at least two skin color-related genes (miR-495-y, miR-543-y, miR-665-z, miR-433-y, and miR-382-x). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of target genes identified noncoding RNA metabolic process as the most significantly enriched GO term, and several metabolic pathways associated with skin color were enriched significantly, such as tyrosine metabolism, histidine metabolism, and vitamin B6 metabolism. Quantitative real-time PCR of selected mRNAs and miRNAs validated the reliability of the integrated analysis. This study provides in-depth insights into the molecular mechanism of skin color variation between WR and YR, which will accelerate the genetic selection and breeding of rainbow trout with consumer-favored traits. [Display omitted] • Comparative mRNA and miRNA expression was firstly conducted to identify key mRNAs and miRNAs that may be responsible for skin color variation between wild-type and yellow mutant rainbow trout. • Three key positive genes in the formation of xanthophores and several important genes related to melanogenesis were all up-regulated in yellow mutant rainbow trout. • Five miRNAs were identified that target at least two skin color related genes. • Several metabolic pathways related to skin color were significantly enriched. [ABSTRACT FROM AUTHOR]