Macklis J D, Madison R D
Department of Neurology, Harvard Medical School, Children's Hospital, Boston, MA 02115.
J Neurosci Methods. 1990 Jan;31(1):43-6. doi: 10.1016/0165-0270(90)90007-3.
We describe a visual assay of neuronal electrophysiologic status for use with cultured neurons, based on the exclusion of propidium iodide (PI) by intact cellular membranes. We use this fluorescent dye, which binds to nucleic acids, at concentrations suitable for long-term exposure to neurons without toxicity. We correlate the progressive loss of resting membrane potential and the progressive inability to generate stimulated action potentials by cultured mouse dorsal root ganglion neurons with increasing incorporation of PI. The scoring system used to gauge incorporation of PI is rapid and highly reproducible using a standard fluorescence microscope. Applications exist for studies of neuronal toxicity, survival, and electrophysiology in vitro.
我们描述了一种用于培养神经元的神经元电生理状态视觉检测方法，该方法基于完整细胞膜对碘化丙啶（PI）的排斥。我们使用这种与核酸结合的荧光染料，其浓度适合长期暴露于神经元而无毒性。我们将培养的小鼠背根神经节神经元静息膜电位的逐渐丧失以及产生刺激动作电位的逐渐无能与PI掺入增加相关联。使用标准荧光显微镜，用于评估PI掺入的评分系统快速且高度可重复。该方法可应用于体外神经元毒性、存活和电生理的研究。
