Rhythmic Expression Analysis Of MiRNA In Siniperca Chuatsi Muscle And Liver

Circadian Rhythm is a roughly 24-hour cycle in the behavioral, physiological or biochemical processes of the periodic variation law. It plays an important role in determining the sleeping and feeding patterns of all animals, and also related with brain wave activity, hormone production, cell regeneration and other biological activities. Presently investigation of the mechanism of circadian rhythm primarily has been focused on transcriptional level and post-translational modification, and in post-transcriptional also rarely reported. MicroRNA (miRNAs) interact with 3’untranslated region (UTR) elements and targeting regulation mRNA stability or translation, thus play a role in regulating many different biological processes, including growth, reproduction and disease. However, as an important post-transcriptional regulation of miRNA, little is known about the biological function of circadian rhythms in fish and even mammals. Therefore, mandarin fish (Siniperca chualsi) is the experimental subject of my study. We built up mandarin fish red muscle and mix tissue of small RNA libraries, and then use Solexa high-throughput sequencing technologies for small RNA fragment sequencing in the library. To determine the mandarin fish red muscle, white muscle and liver tissue related miRNA 12h:12h light/dark cycle (LD) condition expression by real-time fluorescence quantitative PCR(qRT-PCT) technique. We analyzed miRNAs with daily rhythmic expression, and correlation between the rhythmic expression of miRNAs in the red muscle, white muscle and liver tissue. The results of this study are as follows:1. Using the Solexa high-throughput sequencing technologies, we identified 204 and 217 miRNAs in the RM and Mix libraries, including two new miRNAs.2. To display circadian rhythms in the red and white muscle, We have selected 42 miRNAs from the red muscle library. Rhythmicity analysis revealed 21 miRNAs with daily rhythmic expression in red muscle tissue, high expression of 10 miRNAs, including miR-206, miR-107, miR-133 and so on, relatively low expression of 11 miRNAs, including miR-29, miR-31, miR-375 and so on.Most of the miRNAs has positive correlation with each other, muscle-specific gene miR-133 and miR-206 has a strongly positive correlation. But miR-499 negative correlated with part of miRNA expression. Rhythmicity analysis revealed 35 miRNAs with daily rhythmic expression in white muscle tissue, high expression of 18 miRNAs, including miR-1, miR-10, miR-21 and so on, relatively low expression of 17 miRNAs, including miR-338, miR-31, miR-93 and so on. Most of the miRNAs has positive correlation with each other, muscle-specific gene miR-133 and miR-206 has a strongly positive correlation. But miR-499 negative correlated with part of miRNA expression. In white muscle tissue rhythm miRNAs, many miRNA has positive correlation with each other. Promoting cell differentiation of the miR-1 and miR-206 positive correlated with miR-133. But miR-499 and miR-375 negative correlated with part of miRNA expression.3. In the liver, we have selected 23 related miRNAs of liver metabolism or diseases to study the expression of circadian. The 14 miRNAs which miR-26a, miR-122, miR-16, miR-33a, miR-200b, miR-221, miR-155, miR-181b, miR-10, miR-21, miR-99b, miR-192, miR-194 and miR-146a displayed circadian rhythmic expression. Which rhythm there is a positive correlation between part of the rhythm of miRNA.