Dissociated neuronal culture expressing ionotropic odorant receptors as a hybrid odorant biosensor--proof-of-concept study.
Artificial odorant sensors generally perform poorer than olfactory systems in living organisms. The excellent performances of living odorant systems are achieved by the molecular recognition abilities of odorant receptors and the neuronal information processing that follows. To take advantages of this, here we propose a novel hybrid odorant biosensor by means of expressing ionotropic odorant receptors of insects into dissociated neuronal cultures of rodents. This combination of materials brings significant advantages such as easy functional expression, prolonged lifetime, and an ability to amplify the weak ionic currents of odorant receptors. In the present work, pheromone receptors and co-receptors of silkmoth, i.e., BmOR1 and BmorOrco, were expressed in neuronal cultures via liposome transfection. Consequently, BmOR1 and BmorOrco were co-expressed in 8% of neuronal cells, and both receptors were co-localized on a cell membrane. In Ca++ imaging experiments, synchronous increase of calcium signals at the presentation of BOL was found in both transfected cells and non-transfected cells in a dose-dependent manner. These results provide the proof-of-concept of the proposed hybrid odorant biosensor.
Research Center for Advanced Science and Technology, The University of Tokyo, Komaba 4-6-1, Tokyo 153-8904, Japan.
Norio
N
Tanada
Takeshi
T
Sakurai
Hidefumi
H
Mitsuno
Douglas J
DJ
Bakkum
Ryohei
R
Kanzaki
Hirokazu
H
Takahashi
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D012831:Bombyx; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D009812:Odorants / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D018035:Receptors, Odorant / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D043563:Receptors, Pheromone / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction
D018035:Receptors, Odorant; D043563:Receptors, Pheromone; D002118:Calcium