肌肉型乙酰胆碱受体在HEK293细胞上的表达

目的　利用受体克隆技术 ,建立骨骼肌细胞乙酰胆碱受体 (m nAChR)的实验模型 ,以便进行各种药理研究。方法　应用分子生物学技术将编码小鼠m nAChR的α、β、γ、δ、ε亚基之cDNA分别重组于真核细胞表达质粒pcDNA3 1+ 上 ,用脂质体转染技术 ,将重组后的pcDNA3 1+ 导入HEK2 93细胞 ,使其细胞膜上表达胚胎型乙酰胆碱受体 (γ nAChR)和成年型乙酰胆碱受体 (ε nAChR) ,这样建立了两种m nAChR亚型的实验模型。应用全细胞膜片钳技术测量转染细胞对该受体的内生配基乙酰胆碱的反应。结果　乙酰胆碱可激发转染细胞产生一内向电流 ,电流的大小与乙酰胆碱呈浓度依赖性。结论　HEK2 93细胞在转染后 2 4～ 72h内 ,其细胞膜上表达γ nAChR或ε nAChR ,用于进行各种药理实验。

Transiently expressed muscle nicotinic acetylcholine receptors in HEK293 cells

Aim Expressing muscle acetylcholin e receptors in mammallian cells using receptor clone technique.Methods The cDNA coding for the α,β,δ, γ,ε-subunits of the mouse muscle nAC hR in pSP65, pSP64,pBS SK(-) are subcloned into pcDNA3.1+ by gene recobinant technique and then transfect HEK293 cells using lipofection technique.The γ-n AChR and ε-nAChR are transiently expressed in HEK293 cells membrane. Recording the response of transfected HEK293 cells to acetylcholine by using the whole- cell clamp technique.Results Transfected HEK293 cells may produ ce a inward current as appling acetylcholine.The current values are acctylcholin e-concentration-dependent.Conclusion γ-nAChR or ε-nAChR i s expressed successfully in HEK293 cells.