Simard J M, Song Y, Tewari K, Dunn S, Werrbach-Perez K, Perez-Polo J R, Eisenberg H M
Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston 77550.
J Neurosci Res. 1993 Feb 1;34(2):170-8. doi: 10.1002/jnr.490340204.
Ionic channels in human cortical neurons have not been studied extensively. HCN-1 and HCN-1A cells, which recently were established as continuous cultures from human cortical tissue, have been shown by histochemical and immunochemical methods to exhibit a neuronal phenotype, but expression of functional ionic channels was not demonstrated. For the present study, HCN-1 and HCN-1A cells were cultured in Dulbecco's modified Eagle's medium with 15% fetal calf serum, in some cases supplemented with 10 ng/ml nerve growth factor, 10 microM forskolin, and 1 mM dibutyryl cyclic adenosine monophosphate to promote differentiation. Cells or membrane patches were voltage clamped using conventional patch clamp techniques. In HCN-1A cells, we identified a tetrodotoxin-sensitive Na+ current, two types of Ca2+ channel current, including L-type current and a second type that in some respects resembled N-type current, and four types of K+ current, including a delayed outward rectifier that showed voltage-dependent inactivation, two types of noninactivating Ca(2+)-activated K+ channels with slope conductances of 146 and 23 pS (K+i/K+o 145 mM/5 mM), and less frequently, a noninactivating, intermediate conductance channel that was not sensitive to internal Ca2+. When HCN-1A cells were examined after 3 days of exposure to differentiating agents, pronounced morphological changes were evident but no differences in ionic currents were apparent. HCN-1 cells also exhibited K+ and Ca2+ channel currents, but Na+ currents were not detected in these cells.(ABSTRACT TRUNCATED AT 250 WORDS)
人类皮层神经元中的离子通道尚未得到广泛研究。HCN - 1和HCN - 1A细胞是最近从人类皮层组织建立的连续培养细胞系,通过组织化学和免疫化学方法已显示出具有神经元表型,但尚未证明其功能性离子通道的表达。在本研究中,HCN - 1和HCN - 1A细胞在含有15%胎牛血清的杜氏改良 Eagle培养基中培养,在某些情况下补充10 ng/ml神经生长因子、10 μM福斯高林和1 mM二丁酰环磷酸腺苷以促进分化。使用传统膜片钳技术对细胞或膜片进行电压钳制。在HCN - 1A细胞中,我们鉴定出一种对河豚毒素敏感的Na⁺电流、两种类型的Ca²⁺通道电流,包括L型电流和在某些方面类似于N型电流的第二种类型,以及四种类型的K⁺电流,包括一种表现出电压依赖性失活的延迟外向整流器、两种斜率电导分别为146和23 pS(细胞内/细胞外K⁺浓度为145 mM/5 mM)的非失活Ca²⁺激活K⁺通道,以及较不常见的一种对细胞内Ca²⁺不敏感的非失活中间电导通道。当在暴露于分化剂3天后检查HCN - 1A细胞时,明显的形态学变化很明显,但离子电流没有明显差异。HCN - 1细胞也表现出K⁺和Ca²⁺通道电流,但在这些细胞中未检测到Na⁺电流。(摘要截短于250字)