乙酰胆碱受体亚型α3β2的表达研究

Study on Expression of Nicotinic Acetylcholine Receptor α3β2 Subtype

烟碱乙酰胆碱受体(nAChRs)是一类重要的配体门控离子通道,广泛分布于肌肉、中枢和外周神经系统。这类受体与学习、感觉、神经紊乱和一些脑部疾病相关。研究nAChRs结构与功能对于治疗相关疾病十分重要。目的:建立nAChRs亚型α3β2在非洲爪蟾卵母细胞上的表达模型。方法:通过体外转录获得α3和β2亚基基因的cRNA,二者等量混合后注射到非洲爪蟾卵母细胞内,通过双电极电压钳技术,利用乙酰胆碱(ACh)刺激,检测所表达的受体是否具有配体门控离子通道的功能。结果:α3β2 nAChRs亚型成功在爪蟾卵母细胞表达,表达率80%,并具有很好的ACh配体门控敏感性。结论:研究结果为针对α3β2 nAChRs亚型的分子探针和新药筛选提供了模型和基础。

Nicotinic acetylcholine receptors （nAChRs） are important ligand- gated ion channels, which are widely distributed in the mus- cle, central nervous system and peripheral nervous system. These receptors mediate learning, feeling, nerve disorder and some severe pathol- ogies of the brain. Study on the structure and function of nAChRs is very important for the treatment of related diseases. Objective：The α3β2 nAChRs subtype expression model were constructed in Xenopus oocytes. Method：Both cRNAs of α3 and β2 subunit were acquired by in vitro transcription, then the cRNAs were mixed in equal ratio and injected into Xenopus laevis oocytes, two - electrode voltage clamp recorded inward currents elicited by acetylchofine （ACh） with different concentrations to detect whether the receptors have the ligand gated ion channel function. Result：The α3β2 nAChRs subtype were successfully expressed in oocytes membrane, the expression rate is 80% , and the receptor have excellent sensitivity of ligand gated ion channel function. Conclusion ：The results will provide an efficient model and basis for screening α3β2 subtype specific probes and new drugs.