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

阐明鱼类骨骼肌无限生长的分子机制

基本信息

批准号：
18F18389
负责人：
浅川修一
金额：
$ 1.41万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2018
资助国家：
日本
起止时间：
2018-11-09 至 2021-03-31
项目状态：
已结题

项目摘要

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.

与斑马鱼和大鼠之间的衰老相关的DEG配置文件的比较分析表明，既保守，明显不同的衰老现象。我们的研究结果表明，鱼类和哺乳动物的衰老过程既有共同的，也有不同的。从3个年龄组罗非鱼血清中提取外泌体RNA，利用Agilent 2100生物分析仪分析外泌体小RNA的质量和大小分布，清晰显示了miRNA的picks。在确认RNA质量后，成功构建了3个年龄组的12个外泌体小RNA文库，并通过HiSeq 2500测序，调查miRNA的多样性。在去除已知RNA和基因组未映射读段后，本研究中剩余1426万个读段用于miRNA鉴定。为了鉴定不同年龄组罗非鱼外泌体中的已知miRNA，我们将我们的数据集与miRBase 22.0中的已知miRNA进行了比较。在不允许序列之间有超过一个错配的情况下，鉴定了80种已知的miRNA。为了研究miRNAs对衰老的影响，将预测所选签名miRNAs的靶基因以及三个年龄组中最丰富的miRNAs。将从Gene Ontology数据库中进行靶基因的GO注释，p值阈值为0.05，用于其功能分析。计划中的研究结果将有助于确定外泌体相关miRNA的新的和新颖的作用，这些miRNA首次参与硬骨鱼的衰老。