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

作　　者：施西子 毛楷林 朱晓鹏[1] 长孙东亭[1] 罗素兰[1] SHI Xizi;MAO Kailin;ZHU Xiaopeng;ZHANGSUN Dongting;LUO Sulan(Key Laboratory of Tropical Biological Resources of Ministry of Education/Key Lab for Marine Drugs of Haikou/School of Life and Pharmaceutical Sciences, Hainan University, Haikou 570228,China)

机构地区：[1]海南大学热带生物资源教育部重点实验室/海口市海洋药物重点实验室/海南大学生命科学与药学院,海口570228

出　　处：《华南师范大学学报（自然科学版）》2020年第3期70-77,共8页Journal of South China Normal University(Natural Science Edition)

基　　金：国家自然科学基金项目(81872794,41966003);高等学校学科创新引智计划资助项目(D20010)。

摘　　要：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.

关 键 词：α6/α3β2β3乙酰胆碱受体 非洲爪蟾卵母细胞 双电极电压钳 表达优化 

分 类 号：Q812[生物学—生物工程] Q786