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Am J Physiol Cell Physiol (August 10, 2005). doi:10.1152/ajpcell.00182.2005
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Submitted on April 19, 2005
Accepted on August 4, 2005

Quantification of Gap Junction Selectivity

Jose F Ek-Vitorin1 and Janis M Burt1*

1 Physiology, University of Arizona, Tucson, Arizona, USA

* To whom correspondence should be addressed. E-mail: jburt{at}u.arizona.edu.

Gap junctions, essential for functional coordination and homeostasis within tissues, permit the direct intercellular exchange of small molecules. The abundance and diversity of this exchange depends on the number and selectivity of the comprising channels, and the transjunctional gradient for and chemical character of the permeant molecules. Limited knowledge of functionally significant permeants and poor detectability of those few known have made it difficult to define channel selectivity. Presented here is a multi-faceted approach to quantification of gap junction selectivity that includes determining the rate constant for intercellular diffusion of a fluorescent probe (k2-DYE) and junctional conductance (gj) for each junction studied, such that the selective permeability (k2-DYE/gj) for dyes with differing chemical characteristics or junctions with differing connexin compositions (or treatment conditions) can be compared. In addition, selective permeability can be correlated with single channel conductance ({gamma}j) when this parameter is also measured. Our measurement strategy is capable of detecting: 1) rate constants and selective permeabilities that differ over three orders of magnitude, and 2) acute changes in that rate constant. With this strategy we show that: 1) the selective permeability of Cx43 junctions to a small cationic dye varied across two orders of magnitude, consistent with the supposition that the various channel configurations adopted by Cx43 display different selective permeabilities; and 2) the selective permeability of Cx37 vs. Cx43 junctions was consistently and significantly lower.




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