Chemical gating of heteromeric and heterotypic gap junction channels

Gap junction channels contain two hemichannels (connexons), each being a connexin (Cx) hexamer. In cells expressing multiple connexins, heteromeric connexons are believed to form, whereas cell pairs expressing different connexins generate heterotypic channels. To define gating behavior of heteromeric and heterotypic channels, CO2-induced gating was tested in Xenopus oocyte pairs expressing Cx32, or 5R/N (Cx32 mutant), as well as in pairs in which one oocyte (mx) expressed a 50/50 mixture of Cx32 and 5R/N and the other either the mixture (mx), Cx32 (32) or 5R/n (R/N). In 5R/N, replacement of 5 C-terminus arginines with asparagines greatly increased CO2 sensitivity. In response to 3 and 15 min CO2 exposures, junctional conductance (Gj) decreased to 85% and 47%, in 32-32 pairs, and to 7% and 0.9%, in R/N-R/N pairs, respectively. In mx-mx and mix-32 pairs, Gj decreased to similar values (33% and 35%, respectively) with 15 min CO2. The sensitivity of mx-R/N pairs was similar to that of heterotypic 32-R/N pairs, as Gj dropped to 36% and 38%, respectively, with 3 min CO2. Monoheteromeric (mx-32 and mx-R/N) and biheteromeric (mx-mx) channels behaved as if Cx32 were dominant, suggesting that hemichannel sensitivity is not an average of the sensitivities of its connexin monomers. In contrast, heterotypic channels behaved as if the two hemichannels of a cell-cell channel had no influence on each other.