title: Real-time detection of L-glutamate released from C6 glioma cells using a modified enzyme-luminescence method.
abstract: There is an increasing interest in new strategies to detect neurotransmitters released from nerve cells in real time for brain science, drug assessment, and so on. Previously we reported real-time monitoring of dopamine release from nerve model cells by enzyme-catalyzed luminescence measurement with tyramine oxidase and peroxidase. In the present study, the system was modified with glutamate oxidase instead of tyramine oxidase to detect L-glutamate sensitively ( approximately 10 nM) and rapidly with high temporal resolution (<1 s). We applied this modified method successfully to perform real-time monitoring of L-glutamate release from brain model cell (C6 glioma cell) using a luminescence plate reader upon stimulation with high concentration of KCl (>10 mM) or 5-hydroxytryptamine (>1 microM). The measurement solution was not toxic and therefore the L-glutamate release from the cell was measured by the second stimulation after exchanging the measurement solution. We conclude that the developed monitoring system is suitable for real-time detection of dynamic L-glutamate release from nerve cells in vitro and will be suitable for application in assessment of drugs acting on the nervous system.
authors.affiliation: Department of Advanced Nano and Biosciences, Graduate School of Innovative Life Science, University of Toyama, 3190 Gofuku, Toyama 930-8555. Japan. zhossain@ucalgary.ca
authors.forename: S M
authors.initials: SM
authors.lastname: Zakir Hossain
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Shinohara
authors.forename: Feifei
authors.initials: F
authors.lastname: Wang
authors.forename: Hiromi
authors.initials: H
authors.lastname: Kitano
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D005910:Glioma / Q000378:metabolism*; D018698:Glutamic Acid / Q000032:analysis* / Q000378:metabolism; D049449:Luminescence; D010088:Oxidoreductases / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000494:pharmacology; D018490:Serotonin Agents / Q000494:pharmacology; D013268:Stimulation, Chemical; D013997:Time Factors
chemical_list: D018490:Serotonin Agents; D012701:Serotonin; D018698:Glutamic Acid; D011189:Potassium Chloride; D010088:Oxidoreductases