Molecular cloning, distribution and functional analysis of the NAV1.6.: Voltage-gated sodium channel from human brain
Molecular cloning, distribution and functional analysis of the NAV1.6.: Voltage-gated sodium channel from human brain
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DOI：
10.1016/s0169-328x(02)00188-2
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发表时间：
2002-06-30
期刊：
影响因子：
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通讯作者：
Clare, JJ
中科院分区：
文献类型：
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作者：
Burbidge, SA;Dale, TJ;Clare, JJ
我们已经克隆并表达了全长人Na(v)1.6钠通道cDNA。Northern分析表明，hNa(v)1.6基因与其啮齿动物同源基因一样，在成年大脑中大量表达，但在包括心脏和骨骼肌在内的其他组织中不表达。在成年大脑中，hNa(v)1.6 mRNA广泛表达，在小脑、枕极和额叶中水平尤其高。当在人胚胎肾细胞（HEK293）中稳定表达时，发现hNa(v)1.6通道的生物物理特性与人Na(v)1.2和Na(v)1.3通道非常相似[《欧洲神经科学杂志》12（2000）4281 - 4289；《普弗吕格尔斯档案》441（2001）425 - 433]。仅观察到相对细微的差异，例如在门控的电压依赖性方面。与hNa(v)1.3通道一样，当在HEK293细胞中表达时，hNa(v)1.6产生具有显著持续成分的钠电流。这些持续电流与报道的大鼠Na(v)1.2通道的电流相似[《神经元》19（1997）443 - 452]，尽管它们不依赖于G蛋白βγ亚基的过表达。这些数据与Na(v)1.6通道可能产生在小脑浦肯野神经元中观察到的持续电流的观点一致[《神经科学杂志》17（1997）4157 - 4536]。然而，在我们的hNa(v)1.6细胞系中，我们无法检测到在浦肯野细胞中也有描述的再生电流。尽管Na(v)1.6通道已被认为与产生这些再生电流有关[《神经元》19（1997）881 - 891]，但我们的数据表明，这可能需要浦肯野神经元中存在而HEK293细胞中不存在的其他因子对Na(v)1.6的α亚基进行修饰。（C）2002爱思唯尔科学出版社B V版权所有。
We have cloned and expressed the full-length human Na(v)1.6 sodium channel cDNA. Northern analysis showed that the hNa(v)1.6 gene, like its rodent orthologues, is abundantly expressed in adult brain but not other tissues including heart and skeletal muscle. Within the adult brain, hNa(v)1.6 mRNA is widely expressed with particularly high levels in the cerebellum, occipital pole and frontal lobe. When stably expressed in human embryonic kidney cells (HEK293), the hNa(v)1.6 channel was found to be very similar in its biophysical properties to human Na(v)1.2 and Na(v)1.3 channels [Eur. J. Neurosci. 12 (2000) 4281-4289; Pflugers Arch. 441 (2001) 425-433]. Only relatively subtle differences were observed, for example, in the voltage dependence of gating. Like hNa(v)1.3 channels, hNa(v)1.6 produced sodium currents with a prominent persistent component when expressed in HEK293 cells. These persistent currents were similar to those reported for the rat Na(v)1.2 channel [Neuron 19 (1997) 443-452], although they were not dependent on over-expression of G protein betagamma subunits. These data are consistent with the proposal that Na(v)1.6 channels may generate the persistent currents observed in cerebellar Purkinje neurons [J. Neurosci. 17 (1997) 4157-4536]. However, in our hNa(v)1.6 cell line we have been unable to detect the resurgent currents that have also been described in Purkinje cells. Although Na(v)1.6 channels have been implicated in producing these resurgent currents [Neuron 19 (1997) 881-891], our data suggest that this may require modification of the Na(v)1.6 a subunit by additional factors found in Purkinje neurons but not in HEK293 cells. (C) 2002 Elsevier Science B V All rights reserved.