Liu R H, Morassutti D J, Whittemore S R, Sosnowski J S, Magnuson D S
Department of Neurological Surgery, University of Louisville School of Medicine, Kentucky 40202, USA.
Exp Neurol. 1999 Jul;158(1):143-54. doi: 10.1006/exnr.1999.7078.
Growth factor-expanded neural precursor cells isolated from the mammalian central nervous system can differentiate into neurons and glia. Although the morphological and neurochemical development of these neural precursor cells has been investigated, little attention has been paid to their electrophysiology. This study examined the electrophysiological properties of neurons and glia derived from neural precursor cells isolated from the adult rat spinal cord (SC) and subventricular zone (SVZ). Cells were cultured in medium containing epidermal growth factor and/or fibroblast growth factor-2. After at least two passages, spheres of neural precursor cells were plated on coated coverslips and maintained in culture for up to 6 weeks. Whole-cell patch recordings were made using standard current clamp techniques. Immature action potentials were observed within hours of plating for both SC and SVZ cells. Input resistance and time constants decreased over the first week after plating and no further changes were found at later times. At similar times following plating, however, SVZ cells had a lower input resistance and shorter time constant compared to SC cells. SVZ cells also had higher resting membrane potentials and smaller after hyperpolarizations than those of SC cells, despite no significant difference in the amplitude of action potentials. Neither the SC nor the SVZ cells were capable of eliciting more than a single action potential in response to injected current. While all SC cells tested were depolarized by glutamate, the response of SVZ cells to glutamate varied considerably. This study revealed that neural precursor cells from SC and SVZ differ in both active and passive membrane properties. It appears also that the electrophysiological development of SC and SVZ precursor-derived neurons is incomplete under the conditions used. These observations suggest that the neural precursor cells from different anatomical locations may be physiologically diverse and may exhibit some differences in commitment toward neuronal or glial phenotypes.
从哺乳动物中枢神经系统分离出的生长因子扩增神经前体细胞可分化为神经元和神经胶质细胞。尽管已经对这些神经前体细胞的形态和神经化学发育进行了研究,但对其电生理学关注甚少。本研究检测了从成年大鼠脊髓(SC)和脑室下区(SVZ)分离出的神经前体细胞所衍生的神经元和神经胶质细胞的电生理特性。细胞在含有表皮生长因子和/或成纤维细胞生长因子-2的培养基中培养。至少传代两次后,将神经前体细胞球接种在包被的盖玻片上,并在培养中维持长达6周。使用标准电流钳技术进行全细胞膜片钳记录。接种后数小时内,SC和SVZ细胞均观察到未成熟动作电位。接种后第一周内输入电阻和时间常数降低,之后未发现进一步变化。然而,在接种后的相似时间,与SC细胞相比,SVZ细胞的输入电阻较低,时间常数较短。尽管动作电位幅度无显著差异,但SVZ细胞的静息膜电位也高于SC细胞,超极化后电位较小。无论是SC细胞还是SVZ细胞,对注入电流的反应都不能引发超过单个动作电位。虽然所有测试的SC细胞都被谷氨酸 depolarized,但SVZ细胞对谷氨酸的反应差异很大。本研究表明,来自SC和SVZ的神经前体细胞在主动和被动膜特性方面均存在差异。在所用条件下,SC和SVZ前体衍生神经元的电生理发育似乎也不完全。这些观察结果表明,来自不同解剖位置的神经前体细胞在生理上可能存在差异,并且在向神经元或神经胶质细胞表型的分化上可能表现出一些差异。