α6/α3β2β3乙酰胆碱受体在非洲爪蟾卵母细胞中的表达及其条件优化

The Expression ofα6/α3β2β3 Acetylcholine Receptor in Xenopus laevis Oocytes and the Conditions for Its Optimization

为建立有效的α6/α3β2β3 nAChRs(α6/α3β2β3,或简写为α6β2*,*代表其他亚基)体外重组表达实验模型,利用非洲爪蟾卵母细胞表达体系,采用显微注射RNA的方法,利用双电极电压钳技术检测电流,对α6/α3β2β3 nAChRs亚型进行体外重组表达研究,并对其表达条件进行优化,获得了α6/α3β2β3 nAChRs优化的重组表达体系.烟碱型乙酰胆碱受体(nAChRs)是一类重要的配体门控离子通道,其亚型众多且结构相似,但各亚型的生理学和药理学功能截然不同.天然的α6β2*nAChRs很难体外表达,体外表达模型的表达电流非常小.因此,本实验拟研究优化该亚型的表达,进行优化后的体外表达模型,产生的激发电流显著提高,为以该亚型为靶点的新药筛选提供实验模型和基础.

Nicotinic acetylcholine receptors(nAChRs),an important kind of ligand-gated ion channels with many subtypes,are similar in structures but different in physiological and pharmacological functions.Naturalα6β2*nAChRs are unlikely to be expressed in vitro and their expression currents are quite small.In order to establish an effective experimental model ofα6/α3β2β3 nAChRs in vitro,the expression system of Xenopus laevis oocytes was used to express theα6/α3β2β3 nAChRs subtype with the method of micro injection of RNA and detection with two-electrode voltage clamp.The expression conditions were optimized to obtain a better and more efficient expression system forα6/α3β2β3 nAChRs.The current generated by the expressed receptor was significantly increased.This may provide an experimental model and basis for the screening of new drugs targeting this subtype.