A Biosensing System Based on Extracellular Potential recording of Ligand-Gated Ion Channel Function Overexpressed in Insect cells
Date
2003
Authors
Haruyama, T.
Bongsebandhu-Phubhakdi, S.
Nakamura, I.
Mottershead, D.
Keinanen, K.
Kobatake, E.
Aizawa, M.
Editors
Advisors
Journal Title
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Volume Title
Type:
Journal article
Citation
Analytical Chemistry, 2003; 75(4):918-921
Statement of Responsibility
Tetsuya Haruyama, Saknan Bongsebandhu-Phubhakdi, Ibuki Nakamura, David Mottershead, Kari Keinänen, Eiry Kobatake, and Masuo Aizawa
Conference Name
Abstract
We have used outer cell potential measurement to record agonist-dependent cellular responses in cells engineered to express ligand-gated ion channels and grown on a microelectrode surface. Application of glutamate, a natural agonist, induced a complex and robust potentiometric response in cells expressing homomeric GluR-D glutamate receptor, but not in nonexpressing control cells. The response consisted of an initial decrease in outer potential followed by a transient increase and was not obtained for other amino acids devoid of agonist activity at glutamate receptors. Furthermore, the pharmacological agonist of the GluR-D receptor, kainate, also produced the potentiometric response whereas 6-cyano-7-nitroquinoxaline-2,3-dione, a competitive antagonist, was not active in itself but attenuated the responses to glutamate. The time course of the measured changes was slow, which may be partially due to the ligand being applied by free diffusion but may also reflect a contribution by secondary changes in the behavior of the cells. This novel approach should be applicable to other ligand-gated ion channels and holds promise as a cell-based biosensor for high-throughput drug screening and other applications.
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Dissertation Note
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Copyright © 2003 American Chemical Society