Elucidation of molecular mechanism of indeterminate growth of skeletal muscle in fish

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Comparative analysis of DEG profiles associated with aging between zebrafish and rat showed both conserved and clearly different aging phenomena. Our results indicate both common and different aging profiles between fish and mammals.Exosomes were isolated with differential centrifugation and commercial kit. After isolation of RNA from the serum exosome of three age-group of tilapia, we analyzed the quality and size distribution of the exosomal small RNA by using Agilent 2100 Bioanalyzer, which clearly showed the miRNA picks. After confirming the RNA quality, twelve exosomal small RNA libraries from the three age groups were successfully constructed and sequenced by HiSeq 2500 sequencing to survey the miRNA diversity. After removing known RNAs and genome unmapped reads, 14.26 million reads were remaining for miRNA identification in this study. To identify known miRNAs in the exosome of different age-group tilapia, we compared our dataset with the known miRNAs in miRBase 22.0. Allowing no more than one mismatch between sequences, 80 known miRNAs were identified. For the purpose of investigating the influence of miRNAs on the aging, the target genes of the selected signature miRNAs as well as most abundant miRNAs in the three age-groups will be predicted. GO annotation of target genes will be conducted from Gene Ontology database with p-value threshold of 0.05 for their function analysis. The outcomes of the planned research would facilitate to identify the new and novel roles of exosome-associated miRNAs, which are involved in aging in the teleost for the first time.