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Differential expression of two-pore domain potassium channels in rat cerebellar granule neurons

机译:大鼠小脑颗粒神经元中两孔结构域钾通道的差异表达

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Two pore domain potassium (K2P) channels are mostly present in the central nervous system (CNS) where they play important roles in modulating neuronal excitability. K2P channels give rise to background K+ currents (IKso) a key component in setting and maintaining the resting membrane potential in excitable cells. Here, we studied the expression and relative abundances of K2P channels in cerebellar granule neurons (CGNs), combining molecular biology, electrophysiology and immunologic techniques. The CGN IKso so was very sensitive to external pH, as previously reported. Quantitative determination of mRNA expression level demonstrated the existence of an accumulation pattern of transcripts in CGN that encode K2P9 > K2P1 > K2P3 > K2P18 > K2P2 = K2P10 > K2P4 > K2P5 subunits. The presence of the major K2P subunits expressed was then confirmed by Western blot and immunofluorescence analysis, demonstrating robust expression of K2P1 (TWIK-1), K2P3 (TASK-1), K2P9 (TASK-3) and K2P18 (TRESK) channel protein. Based, on these results, it is concluded that K2P1, -3, -9 and -18 subunits represent the majority component of IKso current in CGN. (C) 2014 Elsevier Inc. All rights reserved.
机译:两个孔结构域钾(K2P)通道主要存在于中枢神经系统(CNS)中,在调节神经元兴奋性中起重要作用。 K2P通道产生背景K +电流(IKso),这是设置和维持可激发细胞中静止膜电位的关键因素。在这里,我们结合分子生物学,电生理学和免疫学技术研究了小脑颗粒神经元(CGNs)中K2P通道的表达和相对丰度。如先前报道,CGN IKso对外部pH非常敏感。 mRNA表达水平的定量测定表明,CGN中存在编码K2P9> K2P1> K2P3> K2P18> K2P2 = K2P10> K2P4> K2P5亚基的转录本积累模式。然后通过蛋白质印迹和免疫荧光分析确认表达的主要K2P亚基的存在,证明K2P1(TWIK-1),K2P3(TASK-1),K2P9(TASK-3)和K2P18(TRESK)通道蛋白表达稳定。基于这些结果,可以得出结论,K2P1,-3,-9和-18亚基代表了CGN中IKso电流的主要成分。 (C)2014 Elsevier Inc.保留所有权利。

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