Scanning Ion Conductance Microscopy Measurement of Paracellular Channel Conductance in Tight Junctions
Scanning Ion Conductance Microscopy Measurement of Paracellular Channel Conductance in Tight Junctions
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DOI：
10.1021/ac303441n
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发表时间：
2013-04-02
影响因子：
7.4
通讯作者：
Baker, Lane A.
中科院分区：
文献类型：
--
作者：
Chen, Chiao-Chen;Zhou, Yi;Baker, Lane A.
阐明跨跨细胞或细胞旁途径的上皮运输有望推进目前对离子运输的理解，并能够调节对上皮细胞和分子生物学至关重要的细胞连接。在这里，我们展示了一种新的仪器技术，电位扫描离子电导显微镜（P-SICM），利用纳米级移液管区分旁细胞和跨细胞运输过程中的高空间分辨率。该技术是经过验证的明确定义的聚合物膜，然后用于研究Madin-Darby犬肾株II（MDCKII）细胞的野生型和claudin缺陷突变体。由密蛋白-2赋予的细胞旁渗透性被P-SICM捕获，这证明了在亚细胞水平监测表观电导的实用性。
Elucidation of epithelial transport across transcellular or paracellular pathways promises to advance the present understanding of ion transport and enables regulation of cell junctions critical to the cell and molecular biology of the epithelium. Here, we demonstrate a new instrumental technique, potentiometric scanning ion conductance microscopy (P-SICM), that utilizes a 'nanoscale pipet to differentiate paracellular and transcellular transport processes at high spatial resolution. The technique is validated for well-defined polymer membranes and then employed to study wild type and claudin-deficient mutants of Madin-Darby Canine Kidney strain II (MDCKII) cells. Paracellular permeabilities conferred by claudin-2 are captured by P-SICM which demonstrates the utility to monitor apparent conductance at subcellular levels.