Expression of α3β4 Acetylcholine Receptor in Xenopus Laevis Oocytes

To establish a drug screening model with the molecular target of mammal α3β4 acetylcholine receptor(nAChR),rat α3 and β4 nAChR subunits' cRNA were obtained by transcription in vitro,which were mixed with equal amount and then microinjected in the fresh oocytes of Xenopus laevis.The oocytes were incubated in ND96 buffer at 17 ℃ over 24 hours after microinjection.Two-microelectrode voltage clamp(TEVC) was used to detect the acetylcholine(Ach) gated current of α3β4 nAChR.The Ach gated inward current was recorded in the α3 and β4 cRNA injected oocytes,while no current was recorded in both blank oocytes and ddH2O injected oocytes.The results demonstrated that the α3β4 nAChR was expressed in Xenopus laevis oocytes successfully.This can be used as the experimental model to screen bioactive toxins with the drug target of α3β4 nAChR.