Drug assessment based on detection of L-glutamate released from C6 glioma cells using an enzyme-luminescence method.
Monitoring of excitation activity of nerve cells is very useful for not only brain research but also assessment of the effects of various chemicals, including drugs and toxins. We previously reported a novel enzyme-luminescence method for real-time monitoring of l-glutamate release from C6 glioma cells with high levels of sensitivity ( approximately 10 nM) and temporal resolution (<1 s) using a luminescence plate reader. In the present study, we tested the applicability of this novel system for assessment of effects of drugs in vitro. Several drugs (e.g., veratridine and 4-aminopyridine) were administered to C6 glioma cells for inducing glutamate release. Moreover, antagonists of voltage-dependent Ca (2+) channels (e.g., nifedipine, flunarizine, and NiCl 2) and Na (+) channels (e.g., carbamazepine and lidocaine) were applied separately for evaluating the effects of these chemicals on glutamate release from the cells. The combined effect of carbamazepine and lidocaine was also investigated by using our method, and the combined effect was found to be more potent than that of single drug administration. These results indicated that the glutamate release from C6 cells was modulated by these drugs in a way similar to that found by using several conventional analytical techniques. We therefore conclude that the developed monitoring system for real-time detection of dynamic l-glutamate release from cells could be very useful for application to assessment of drugs acting on the nervous system.
Zakir Hossain
S M
SM
Course of Advanced Nano and Biosciences, Graduate School of Innovative Life Science for Education, University of Toyama, 3190 Gofuku, Toyama 930-8555, Japan. zhossain@ucalgary.ca
Shinohara
Hiroaki
H
Kitano
Hiromi
H
D015761:4-Aminopyridine / Q000008:administration & dosage; D000818:Animals; D001932:Brain Neoplasms / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D002220:Carbamazepine / Q000008:administration & dosage; D045744:Cell Line, Tumor; D004798:Enzymes / Q000378:metabolism*; D005444:Flunarizine / Q000008:administration & dosage; D005910:Glioma / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D018698:Glutamic Acid / Q000378:metabolism*; D008012:Lidocaine / Q000008:administration & dosage; D049449:Luminescence; D009532:Nickel / Q000008:administration & dosage; D009543:Nifedipine / Q000008:administration & dosage; D051381:Rats; D014701:Veratridine / Q000008:administration & dosage
D004798:Enzymes; D002220:Carbamazepine; D018698:Glutamic Acid; C022838:nickel chloride; D014701:Veratridine; D009532:Nickel; D008012:Lidocaine; D015761:4-Aminopyridine; D009543:Nifedipine; D005444:Flunarizine