Expression of olfactory receptors in Xenopus oocytes.
The rat olfactory epithelium and the amino acid-sensitive catfish olfactory system have been used as models to study the molecular mechanisms of olfactory transduction. Here we report the functional expression of rat and catfish olfactory receptors in Xenopus oocytes injected with mRNA isolated from the respective tissues. Application of odor ligands to injected oocytes, monitored by two-electrode voltage clamp, activates stimulus-dependent transmembrane currents that reverse direction at about the chloride equilibrium potential. The currents show characteristic secondary oscillations that are presumed to reflect underlying Ca2+ oscillations. Similar ligand-activated membrane currents induced in oocytes after injection of other mRNAs have been shown to be due to activation of endogenous Ca(2+)-activated chloride channels. In summary, our results demonstrate the usefulness of the Xenopus oocyte expression system for cloning and characterization of olfactory receptors in both fish and mammalian species.
Dahmen
N
N
Department of Neurosciences, Roche Institute of Molecular Biology, Nutley, New Jersey 07110.
Wang
H L
HL
Margolis
F L
FL
D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000502:physiology; D002397:Catfishes; D018118:Chloride Channels; D004594:Electrophysiology; D005021:Ethylamines / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000096:biosynthesis; D008565:Membrane Proteins / Q000502:physiology; D009812:Odorants; D009833:Olfactory Pathways / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D009865:Oocytes / Q000648:ultrastructure*; D011719:Pyrazines / Q000494:pharmacology; D011984:Sensory Receptor Cells / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D013268:Stimulation, Chemical; D014981:Xenopus / Q000502:physiology*
D018118:Chloride Channels; D005021:Ethylamines; D008565:Membrane Proteins; D011719:Pyrazines; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium; C016162:triethylamine