Expression of Muscle Acetylcholine Receptors in Xenopus Laevis Oocytes

To establish the expression models of fetal(α1β1δγ) and adult(α1β1δe) muscle acetylcholine receptors(nAChRs) in Xenopus laevis oocytes for drug screening based on these two molecular targets.Each subunit cRNA of the muscle nAChRs was obtained by in vitro transcription,which was microinjected in Xenopus laevis oocytes for expression.The oocytes were incubated in ND96 buffer at 17 ℃ over 24 h after microinjection.Two-microelectrode voltage clamp(TEVC) was used to detect the acetylcholine gated current of α1β1δγ and α1β1δe nAChRs.The α1β1δγ and α1β1δe muscle nAChRs were expressed in Xenopus laevis oocytes successfully.When the ACh concentration of 1 μmol / L,α1β1δγ and α1β1δe receptor currents are small,and can not be used for subsequent testing.The concentration of Ach 5 μmol / L,fully activated receptor α1β1δγ-current reached the maximum 6 025 nA.The Ach concentration of 20 μmol / L,fully activated receptor α1β1δγ-current reached the maximum 2 742 nA,while no current was recorded ddH2O injected oocytes.The establishment of the receptor expression offered the experimental model to screen drugs targeting α1β1δγ and α1β1δe nAChRs.