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Expression and biophysical properties of Kv1 channels in supragranular neocortical pyramidal neurones

机译:Kv1通道在肾上皮新皮质锥体神经元中的表达及其生物物理特性

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摘要

Potassium channels are extremely diverse regulators of neuronal excitability. As part of an investigation into how this molecular diversity is utilized by neurones, we examined the expression and biophysical properties of native Kv1 channels in layer II/III pyramidal neurones from somatosensory and motor cortex. Single-cell RT-PCR, immunocytochemistry, and whole cell recordings with specific peptide toxins revealed that individual pyramidal cells express multiple Kv1 α-subunits. The most abundant subunit mRNAs were Kv1.1 > 1.2 > 1.4 > 1.3. All of these subunits were localized to somatodendritic as well as axonal cell compartments. These data suggest variability in the subunit complexion of Kv1 channels in these cells. The α-dendrotoxin (α-DTX)-sensitive current activated more rapidly and at more negative potentials than the α-DTX-insensitive current, was first observed at voltages near action potential threshold, and was relatively insensitive to holding potential. The α-DTX-sensitive current comprised about 10% of outward current at steady-state, in response to steps from −70 mV. From −50 mV, this percentage increased to ∼20%. All cells expressed an α-DTX-sensitive current with slow inactivation kinetics. In some cells a transient component was also present. Deactivation kinetics were voltage dependent, such that deactivation was slow at potentials traversed by interspike intervals during repetitive firing. Because of its kinetics and voltage dependence, the α-DTX-sensitive current should be most important at physiological resting potentials and in response to brief stimuli. Kv1 channels should also be important at voltages near threshold and corresponding to interspike intervals.
机译:钾通道是神经元兴奋性的极为不同的调节剂。作为研究神经元如何利用这种分子多样性的一部分,我们检查了来自体感和运动皮层的II / III层锥体神经元中天然Kv1通道的表达和生物物理特性。单细胞RT-PCR,免疫细胞化学和具有特定肽毒素的全细胞记录显示,单个锥体细胞表达多个Kv1α亚基。最丰富的亚基mRNA为Kv1.1> 1.2> 1.4> 1.3。所有这些亚基都定位于躯体树突状和轴突细胞区室。这些数据表明这些细胞中Kv1通道的亚基复合体的变异性。与对α-DTX不敏感的电流相比,对α-树突毒素(α-DTX)敏感的电流激活速度更快,并且具有更多的负电势,首先在接近动作电位阈值的电压下观察到,并且对保持电势相对不敏感。响应于-70 mV的阶跃,在稳态下,对α-DTX敏感的电流约占向外电流的10%。从-50 mV起,该百分比增加到约20%。所有细胞均表达α-DTX敏感电流,具有缓慢的失活动力学。在某些细胞中也存在瞬时成分。失活动力学是电压依赖性的,因此在重复点火期间,在尖峰间间隔遍历的电势下,失活很慢。由于其动力学和电压依赖性,对α-DTX敏感的电流在生理静息电位和对短暂刺激的响应中应该是最重要的。 Kv1通道在接近阈值且对应于尖峰间隔的电压下也应很重要。

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