A Tetracysteine-tag and HeLa Cell System for the Dynamic Analysis of the Localization and Gating Properties of a Specific Connexin Isoform

Abstract

Cell-cell communication in animals is predominantly conducted via gap junction (GJ) plaque comprising 20 connexin (Cx) isoforms. However the GJ plaques are formed at only limited part of cell-cell interface and such an irregular localization is a current problem. To solve this problem, we developed an experimental platform to analyze the role of each Cx isoform in the regulation of GJ plaque localization and in the intercellular molecular movement. A tetracysteine-tagged Cx expressed in HeLa cell deficient in Cx was found feasible for viable imaging of GJ plaques. Femtoinjection enabled the quantitative analysis of intercellular dye diffusion.