Wang D D, Krueger D D, Bordey A
Department of Neurosurgery and Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06520-8082, USA.
J Neurophysiol. 2003 Oct;90(4):2291-302. doi: 10.1152/jn.01116.2002. Epub 2003 Jun 11.
Previous studies have reported the presence of neuronal progenitors in the subventricular zone (SVZ) and rostral migratory stream (RMS) of the postnatal mammalian brain. Although many studies have examined the survival and migration of progenitors after transplantation and the factors influencing their proliferation or differentiation, no information is available on the electrophysiological properties of these progenitors in a near-intact environment. Thus we performed whole cell and cell-attached patch-clamp recordings of progenitors in brain slices containing either the SVZ or the RMS from postnatal day 15 to day 25 mice. Both regions displayed strong immunoreactivity for nestin and neuron-specific class III beta-tubulin, and recorded cells displayed a morphology typical of the neuronal progenitors known to migrate throughout the SVZ and RMS to the olfactory bulb. Recorded progenitors had depolarized zero-current resting potentials (mean more depolarized than -28 mV), very high input resistances (about 4 GOmega), and lacked action potentials. Using the reversal potential of K+ currents through a cell-attached patch a mean resting potential of -59 mV was estimated. Recorded progenitors displayed Ca2+-dependent K+ currents and TEA-sensitive-delayed rectifying K+ (KDR) currents, but lacked inward K+ currents and transient outward K+ currents. KDR currents displayed classical kinetics and were also sensitive to 4-aminopyridine and alpha-dendrotoxin, a blocker of Kv1 channels. Na+ currents were found in about 60% of the SVZ neuronal progenitors. No developmental changes were observed in the passive membrane properties and current profile of neuronal progenitors. Together these data suggest that SVZ neuronal progenitors display passive membrane properties and an ionic signature distinct from that of cultured SVZ neuronal progenitors and mature neurons.
先前的研究报道了出生后哺乳动物脑室内下区(SVZ)和吻侧迁移流(RMS)中存在神经祖细胞。尽管许多研究已检测了移植后祖细胞的存活和迁移以及影响其增殖或分化的因素,但在近乎完整的环境中,关于这些祖细胞的电生理特性尚无相关信息。因此,我们对出生后第15至25天小鼠包含SVZ或RMS的脑片内的祖细胞进行了全细胞和细胞贴附式膜片钳记录。这两个区域对巢蛋白和神经元特异性III类β-微管蛋白均显示出强免疫反应性,且记录的细胞呈现出已知在整个SVZ和RMS中迁移至嗅球的神经祖细胞的典型形态。记录的祖细胞具有去极化的零电流静息电位(平均比-28 mV更去极化)、非常高的输入电阻(约4 GΩ),且无动作电位。通过细胞贴附式膜片钳利用钾电流的反转电位估计平均静息电位为-59 mV。记录的祖细胞表现出钙依赖性钾电流和四乙铵敏感的延迟整流钾(KDR)电流,但缺乏内向钾电流和瞬时外向钾电流。KDR电流呈现出典型的动力学特征,且对4-氨基吡啶和Kv1通道阻滞剂α-银环蛇毒素也敏感。在约60%的SVZ神经祖细胞中发现了钠电流。在神经祖细胞的被动膜特性和电流特征方面未观察到发育变化。这些数据共同表明,SVZ神经祖细胞呈现出与培养的SVZ神经祖细胞和成熟神经元不同的被动膜特性和离子特征。