Alessandrini Andrea, Gavazzo Paola, Picco Cristiana, Facci Paolo
National Research Council - National Institute for the Physics of Matter, Center nanoStructures and bioSystems at Surfaces - S3 Modena, Italy.
Microsc Res Tech. 2008 Apr;71(4):274-8. doi: 10.1002/jemt.20552.
We report on a novel use of electrochemical scanning force microscopy (SFM) for the investigation of morphological modifications occurring in plasma membranes containing voltage-gated ion channels, on membrane potential variation. Membrane patches of Xenopus laevis oocytes microinjected with exogenous KAT1 cRNA, deposited by a stripping method at the surface of a derivatized gold film in inside-out configuration, have been imaged by SFM in an electrochemical cell. A potentiostat was used to maintain a desired potential drop across the membrane. Performing imaging at potential values corresponding to open (-120 mV) and closed (+20 mV) states for KAT1, morphological differences in localized sample zones were observed. Particularly, cross-shaped features involving a significant membrane portion appear around putative channel locations. The reported approach constitutes the first demonstration of an SPM-based experimental technique suitable to investigate the rearrangements occurring to the plasma membrane containing voltage-gated channels on transmembrane potential variation.
我们报告了一种电化学扫描力显微镜(SFM)的新用途,用于研究含有电压门控离子通道的质膜在膜电位变化时发生的形态改变。通过剥离法将注射了外源性KAT1 cRNA的非洲爪蟾卵母细胞的膜片以反转配置沉积在衍生化金膜表面,在电化学池中用SFM对其进行成像。使用恒电位仪维持跨膜的所需电位降。在对应于KAT1开放(-120 mV)和关闭(+20 mV)状态的电位值下进行成像时,观察到局部样品区域的形态差异。特别是,在假定的通道位置周围出现了涉及显著膜部分的十字形特征。所报道的方法首次证明了一种基于扫描探针显微镜的实验技术,适用于研究含有电压门控通道的质膜在跨膜电位变化时发生的重排。