a voltage-sensitive dye-based assay for the identification of differentiated neurons derived from embryonic neural stem cell cultures的电压特异性dye-based分析识别源于胚胎神经干细胞的分化神经元的文化.pdfVIP

A Voltage-Sensitive Dye-Based Assay for the Identification of Differentiated Neurons Derived from Embryonic Neural Stem Cell Cultures ˜ 1. 2. 3 2 2 Richardson N. Leao , Amilcar Reis , Amanda Emirandetti , Michalina Lewicka , Ola Hermanson , ´ 1* Andre Fisahn 1 Neuronal Oscillations Laboratory, Karolinska Institutet, Stockholm, Sweden, 2 Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden, 3 Developmental Genetics Group, Department of Neuroscience, Uppsala University, Uppsala, Sweden Abstract Background: Pluripotent and multipotent stem cells hold great therapeutical promise for the replacement of degenerated tissue in neurological diseases. To fulfill that promise we have to understand the mechanisms underlying the differentiation of multipotent cells into specific types of neurons. Embryonic stem cell (ESC) and embryonic neural stem cell (NSC) cultures provide a valuable tool to study the processes of neural differentiation, which can be assessed using immunohistochemistry, gene expression, Ca2+-imaging or electrophysiology. However, indirect methods such as protein and gene analysis cannot provide direct evidence of neuronal functionality. In contrast, direct methods such as electrophysiological techniques are well suited to produce direct evidence of neural functionality but are limited to the study of a few cells on a culture plate. Methodology/Principal Findings: In this study we describe a novel method for the detection of action potential-capable neurons differentiated from embryonic NSC cultures using fast voltage-sensitive dyes (VSD). We found that the use of extracellularly applied VSD resulted in a more detailed labeling of cellular processe