| Project/Area Number |
15590095
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | Kyoto University (2004)
National Research Institute for Child Health and Development (2003) |
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Principal Investigator |
HIRASAWA Akira Kyoto University, Graduate School of Pharmaceutical Sciences, Department of Genomic Drug Discovery Sciences, Associate Professor, 薬学研究科, 助教授 (70242633)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2004: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2003: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | orphan receptor / ligand hunting / GPR120 / GLP-1 / enteroendocrine cells / obesity / diabetes / GFP / データベース / 細胞内局在 |
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| Research Abstract |
This research aims to establish a novel functional genomics approach and ligand hunting method for orphan receptor by combining technology of subcellular localization analysis for receptor, signal transduction analysis, and expression profile analysis.
Following three experiments were done.
1.Ligand hunting for orphan receptor GPR120
To identify endogenous ligands for GPR120, we performed receptor internalization assay established in this research project and we found that long-chain free fatty acid evoke specific internalization of GPR120. In agreement with internalization assay, long-chain fatty acids were found to evoke specific rise in [Ca^<2+>]_i and ERK activation.
2.Expression profile analysis for GPR120
To clarify the function of the orphan receptor GPR120 I tried the gene expression profile analysis. Using microarray analysis, I found that GPR120 express abundantly in intestines and enteroendocrine cell line STC-1.
3.Functional analysis for GPR120 in GPR120 natively expressing cell line STC-1
In STC-1 cells, free fatty acids stimulation evokes GLP-1 secretion, rise in [Ca^<2+>]_i and ERK activation. Transfection of RNA_i specific for GPR120, inhibited GLP-1 secretion and [Ca^<2+>]_i rise. These results indicate that fatty ac_id induced Ca^<2+> increase and GLP-1 secretion in STC-1 cell is mediated through GPR120.
The discovery of ligand and physiological role for GPR120 prove the effectiveness of the technique established in this research. The purpose of this research was achieved.
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