Long chain fatty acid receptor function using visualization of mitochondrial function

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In this study, we evaluate the long chain fatty acid receptor function using visualization of mitochondrial function and intracellular Ca^<2+> concentration. Mitochondrial pH is known to reflect its functional viability. However, there has been no useful system to measure the quantitative and serial mitochondrial pH. Recently, a pH of organum has been successfully calculated by using GFP, but still it is not a direct quantitative calculation. Here, we developed the new system to evaluate mitochondrial pH of living cell with fluorescence ratio between two GFP. This pH probe system could evaluate the wide and preferable pH range. We applied insulin-stimulating substance, such as glucose, tolbutamide, arginine and potassium, and measure mitochondrial pH. In the cultured β-cells, the mitochondrial pH decreased immediately by glucose and potassium, and leisurely decreased by arginine and tolbutamide. A basal level of the mitochondrial pH of β-cells has increased by aeration in a cultured medium. However, GPR40 agonist quickly decreases the mitochondrial pH. This result suggested that the signal transduction of GPR40 is differ from the others. Additionally, we evaluate the agonist of GPR120 using Ca2+ visualization in STC-1 cell.