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α6/α3β4乙酰胆碱受体在非洲爪蟾卵母细胞中的表达
引用本文:熊洋,朱晓鹏,吴勇,长孙东亭,罗素兰.α6/α3β4乙酰胆碱受体在非洲爪蟾卵母细胞中的表达[J].热带生物学报,2020,11(4):391-398.
作者姓名:熊洋  朱晓鹏  吴勇  长孙东亭  罗素兰
作者单位:海南大学 生命科学与药学院/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,海口 570228
基金项目:国家自然科学基金;海南省重大科技项目
摘    要:α6/α3β4烟碱型乙酰胆碱受体(α6β4* nAChRs,*代表其他亚基)主要分布于大脑海马区、外周神经节和人源肾上腺嗜铬细胞等,与神经性疼痛和炎症反应、情绪等生理疾病密切相关。本研究拟对大鼠α6/α3β4 nAChR在非洲爪蟾(Xenopus laevis)卵母细胞膜上进行重组表达,并利用双电极电压钳电生理学技术检测该重组受体的功能。在不同浓度的激动剂乙酰胆碱的刺激下,比较了α6/α3和β4亚基的不同配比形成的受体通道开放而产生的电流大小,并用其拮抗剂α?芋螺毒素TxID验证了该受体的敏感性。结果表明,亚基不同配比的α6/α3β4 nAChR均可在非洲爪蟾卵母细胞膜上成功表达,并具有较好的配体门控敏感性。

关 键 词:α6/α3β4烟碱型乙酰胆碱受体    非洲爪蟾卵母细胞    双电极电压钳    重组表达
收稿时间:2020-05-19

Expression of α6/α3β4 Nicotinic Acetylcholine Receptor in Xenopus Oocytes
Affiliation:School of Life and Pharmaceutical Sciences, Key Laboratory of Tropical Biological Resources of Ministry of Education, Key Laboratory for Marine Drugs of Haikou, Hainan University, Haikou, Hainan 570228, China
Abstract:α6/α3β4 nicotinic acetylcholine receptors (α6β4*nAChRs, the asterisk denotes the possible presence of additional subunits) are mainly distributed in the hippocampus, peripheral ganglia and human adrenal chromaffin cells, and are closely related to neuropathic and inflammatory pain, stress, etc. Recombinant expression of rat α6/α3β4 nAChR on Xenopus laevis oocytes was detected by using the two-electrode voltage clamp to record inward currents elicited by acetylcholine at different concentrations in light of α and β subunits at different ratios, and the sensitivity of this receptor was verified by its antagonist α-conotoxin TxID. The results showed that α6β4 nAChR subtypes with different ratios of subunits were successfully expressed in the oocyte membrane, and had good sensitivity of ligand-gated ion channel function.
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