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目的研究α-芋螺毒素TxID对具有不同亚基配比组成的大鼠α3β4乙酰胆碱受体(nAChRs)的阻断活性敏感性。方法利用非洲爪蟾卵母细胞表达大鼠α3β4乙酰胆碱受体亚型,将其亚基α3和β4的cRNA分别以3种不同比例,即1∶1、1∶10或10∶1注射到非洲爪蟾卵母细胞中进行表达,形成不同亚基配比的α3β4受体,分别检测和比较α-芋螺毒素TxID对这些不同亚基配比受体的阻断活性。结果成功表达了α3和β4亚基的3个不同配比的受体,且α-芋螺毒素TxID对它们的敏感性有较大差异,与常规的1∶1α3β4乙酰胆碱受体相比,1∶10α3β4乙酰胆碱受体对TxID的敏感度与之接近,其活性差异在2倍以内,而10∶1α3β4乙酰胆碱受体对TxID的敏感度显著下降,其活性差异超过了5倍。结论α-芋螺毒素TxID对不同亚基配比的α3β4乙酰胆碱受体有不同的敏感性,并能反应该受体的α和β亚基不同的组成方式,这对于研究α3β4乙酰胆碱受体的结构和生理功能具有很重要的指导意义。
Objective To investigate the blocking activity of α-conotoxin TxID on rat α3β4 acetylcholine receptors (nAChRs) with different subunits. Methods The rat α3β4 acetylcholine receptor subtype was expressed by Xenopus laevis oocytes. The cRNAs of its subunits α3 and β4 were respectively injected into African paws at three different ratios (1: 1, 1:10 or 10:1) The expression of α3β4 receptors with different subunit ratios was determined by flow cytometry. The blocking activities of α-conotoxin TxID on these receptors with different subunits were detected and compared. Results Three different receptors of α3 and β4 subunits were successfully expressed, and the sensitivity of Tx-conotoxin TxID to them was quite different. Compared with the conventional 1: 1α3β4 acetylcholine receptor, 1: 1 The sensitivity of 10α3β4 acetylcholine receptor to TxID was close to 2 times, while the sensitivity of 10:1α3β4 acetylcholine receptor to TxID was significantly decreased, and its activity was more than 5 times different. CONCLUSION: The α-conotoxin TxID has different sensitivities to α3β4 acetylcholine receptors with different subunits and can reflect the different compositions of the α and β subunits of this receptor. This is of great significance for studying the structure of the α3β4 acetylcholine receptor And physiological function has very important guiding significance.