Abstract
The initial event in olfactory perception is the binding of odorant molecules to specific receptor proteins in the human nose. The interaction between odorant and receptor initiates olfactory signal transduction that leads to a cation influx and change in the membrane potential of the olfactory sensory neuron. In this study, a microfabricated planar electrode was used to measure the generated membrane potential in a heterologous olfactory system. Human embryonic kidney (HEK)-293 cells expressing the olfactory receptor I7 were transfected with the gustatory cyclic nucleotide gated (CNG) channel to amplify the membrane potential. A microfabricated planar electrode was used to measure the electrical responses of odorant-receptor binding. Stimulation of the olfactory receptor with its specific odorant caused an intracellular Ca2+ influx, which was quantitatively measured using a planar electrode. The extracellular field potential generated by the Ca2+ influx through the CNGgust channel of the cells was approximately 10 mV. This cell-based olfactory biosensor, which uses a microfabricated planar electrode for detection, would be useful for screening specific ligands for binding to orphan olfactory receptors.
Original language | English |
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Pages (from-to) | 2659-2664 |
Number of pages | 6 |
Journal | Biosensors and Bioelectronics |
Volume | 24 |
Issue number | 8 |
DOIs | |
State | Published - 15 Apr 2009 |
Bibliographical note
Funding Information:This work was supported by the Korea Science & Engineering Foundation through the Nano-Bioelectronics and Systems Research Center, Seoul National University, Seoul, Korea. The authors wish to acknowledge Dr. Randall R. Reed (Department of Neuroscience, Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA) for the rho-tag gene; Akihito Yasuoka (Bio-oriented Technology Research Advancement Institution, Saitama, Japan) for pVL-I7, Paul A. Gragrave (J. Hillis Miller Health Center, Department of Ophthalmology, University of Florida College of Medicine, Gainesville, Florida, USA) for the B6-30 cell line, and Dr. Keiko Abe (Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan) for the CNGgust channel gene.
Keywords
- Cell-based olfactory biosensor
- Gustatory cyclic nucleotide gated (CNG) channel
- Human embryonic kidney (HEK)-293
- Olfactory receptor
- Planar electrode