2013 Fiscal Year Annual Research Report
魚類のトリアシルグリセロール代謝の制御機構に関する研究
| Project/Area Number |
13J07839
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| Research Institution | The University of Tokyo |
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Principal Investigator |
王 ルー 東京大学, 大学院農学生命科学研究科, 特別研究員(DC2)
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| Keywords | Gene expression patterns / Metabolic pathways / Next generation sequencing / Fasting / Re-feeding / Medaka |
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| Research Abstract |
To investigate the potential genes related to the triacylglycerol (TAG) metabolism, comprehensive analysis of the gene expression patterns in muscle and liver of medaka in response to fasting and re-feeding were performed using next generation sequencing technique.
The changes in expression values of multiple genes involved in the pathways of glycerolipid metabolism and fatty acid degradation implied that fasting might also induce hepatic TAG biosynthesis, while the enhanced release of free fatty acids from hepatic TAG pools and subsequent β-oxidation of fatty acids might lead to the further consumption of TAG in liver. These gene expression patterns may further explain the changes in hepatic TAG levels in my previous studies.
In contrast, the (β-oxidation in skeletal muscle might be restricted by the down-regulations of acyl-CoA synthetase and oxidase during early fasting. Therefore, the muscular TAG levels in the previous results might be increased associated with the enhanced TAG biosynthesis. However, the induced (β-oxidation of fatty acids and lipolysis of intramuscular TAG pools during 8 day fasting might cause the decrease in TAG levels.
In addition, the protein levels of lipoprotein lipase (LPL) in muscle and liver of medaka were also measured by Western Blotting. However, due to the limited protein amounts of tissues in individual and the relatively lower sensitivity and specificity of LPL antibody, LPL protein could not be detected.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
According to the plan for studies in the last year, the analysis of the gene expression patterns and the measurement of LPL protein levels in medaka tissues have been performed. Especially, several important genes related to TAG metabolism have been detected by next generation sequencing analysis.
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| Strategy for Future Research Activity |
Based on the analysis of gene expression patterns by next generation sequencing, we have got some idea of the changes in the expressions of some important genes related to TAG metabolism. To further clarify the functions of these genes in TAG metabolism in response to fasting and re-feeding, novel tissue-specific transgenic medaka will be developed. Furthermore, the mRNA levels of these genes will be also analyzed by real-time PCR.
In addition, the measurement of LPL protein levels will be improved by pooling medaka tissues, further purification of LPL antibody, and using higher sensitive detection method such as immunofluorescence technique.
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