Real-time detection of dopamine released from a nerve model cell by an enzyme-catalyzed luminescence method and its application to drug assessment.
A real-time observation of neurotransmitter release from a nerve cell is a useful method for not only neuroscience research, but also assessing of the influence of chemicals, including drugs, on the human nervous system. In this study, a more simple and sensitive method for real-time monitoring of dopamine release from a nerve model cell was developed. Highly sensitive detection of dopamine was performed by using tyramine oxidase for dopamine oxidation, which was followed by a luminol luminescence reaction. This enzyme-catalyzed luminescence method was applied to observe dopamine release from the PC12 cell as a nerve model cell upon stimulation with acetylcholine and an acetylcholine receptor agonist. The results demonstrated that the real-time monitoring of the activation of the PC12 cell was easily performed by this method. This method possessed many advantages, such as high sensitivity, rapid measurement and no pretreatment for cells. It might be applied to drug screening and the assessment of harmful influences of food additives and pesticides on the nerves.
Shinohara
Hiroaki
H
Field of Life, Information and System Sciences, Graduate School of Science and Engineering, University of Toyama, Japan. hshinoda@eng.u-toyama.ac.jp
Wang
Feifei
F
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D004246:Dimethylphenylpiperazinium Iodide / Q000494:pharmacology; D004298:Dopamine / Q000032:analysis* / Q000378:metabolism*; D004563:Electrochemistry; D006861:Hydrogen Peroxide / Q000032:analysis; D007202:Indicators and Reagents; D049449:Luminescence; D008165:Luminol / Q000737:chemistry; D008995:Monoamine Oxidase / Q000737:chemistry; D009474:Neurons / Q000378:metabolism*; D018722:Nicotinic Agonists / Q000494:pharmacology; D010084:Oxidation-Reduction; D016716:PC12 Cells; D051381:Rats; D013268:Stimulation, Chemical
D007202:Indicators and Reagents; D018722:Nicotinic Agonists; D004246:Dimethylphenylpiperazinium Iodide; D008165:Luminol; D006861:Hydrogen Peroxide; D008995:Monoamine Oxidase; D000109:Acetylcholine; D004298:Dopamine