Department of Physiology and Biophysics, SUNY-Buffalo, Buffalo, NY 14260, USA.
Lab Chip. 2011 Mar 21;11(6):1096-101. doi: 10.1039/c0lc00350f. Epub 2011 Feb 4.
Transport across gap junction channels (GJCs) between neighboring cells is critical to synchronizing cell's electrical and metabolic activities and maintaining cell homeostasis. Here we present a non-invasive microfluidic method to measure molecular diffusion across GJCs in multiple 1D cell arrays in real time. Using the chip, selective loading of a membrane permeant fluorescence dye (carboxyfluorescein) in Normal Rat Kidney (NRK) cells shows that the dye was able to diffuse through three cells along single cell chains in ∼35 minutes. Application of 100 µM 2-aminoethoxydiphenyl borate (2-APB) reversibly inhibits connexin-43 gap junctions in NRK cells; 0.8 mM 1-heptanol inhibits the diffusion partially. The method offers rapid exchange of reagents, enabling sequential screening of multiple gap junction specific drugs with only one preparation of cells. It is capable of measuring gap junction mediated diffusion between single cells.
细胞间隙连接通道（GJCs）在相邻细胞之间的物质转运对于协调细胞的电和代谢活动以及维持细胞内稳态至关重要。在这里，我们提出了一种非侵入性的微流控方法，可实时测量多个 1D 细胞阵列中 GJCs 的分子扩散。使用该芯片，对正常大鼠肾细胞（NRK）中的膜通透性荧光染料（羧基荧光素）进行选择性加载，结果表明该染料能够在大约 35 分钟内通过三个细胞沿着单细胞链扩散。应用 100µM 2-氨基乙氧基二苯硼酸酯（2-APB）可使 NRK 细胞中的连接蛋白-43 间隙连接可逆抑制；0.8mM 1-庚醇部分抑制扩散。该方法提供了快速的试剂交换，仅通过一次细胞制备即可对多种间隙连接特异性药物进行顺序筛选。它能够测量单细胞之间的间隙连接介导的扩散。
