Enzyme-luminescence method: Tool for evaluation of neuronal differentiation based on real-time monitoring of dopamine release response from PC12 cells

Abstract

We previously developed an easy and sensitive method for real-time monitoring of dopamine release from PC12 cells using a combination of enzymatic oxidation of dopamine and luminal reaction. Here, we applied our enzyme-luminescence method for the evaluation of neuronal differentiation in PC12 cells with NGF, via real-time monitoring of dopamine release response. ATP and muscarine-stimulated dopamine release from PC12 cells was compared before and after differentiation by the luminescence peak intensity responding to agonists' injection. A clear enhancement of dopamine release ability in PC12 cells after differentiation was demonstrated. Therefore, this enhancement in luminescence peak intensity by dopamine release upon agonist stimulation may be a good marker of differentiation in PC12 cells. These results imply that the enzyme-luminescence method might be used as a promising tool for evaluation of the effects of various growth factors and chemicals on neuronal differentiation in neuronal progenitor cells.