Functional characteristics of skate connexin35, a member of the gamma subfamily of connexins expressed in the vertebrate retina

Retinal neurons are coupled by electrical synapses that have been studied e xtensively in situ and in isolated cell pairs. Although many unique gating properties have been identified, the connexin composition of retinal gap ju nctions is not well defined. We have functionally characterized connexin35 (Cx35), a recently cloned connexin belonging to the gamma subgroup expresse d in the skate retina, and compared its biophysical properties with those o btained from electrically coupled retinal cells. Injection of Cx35 RNA into pairs of Xenopus oocytes induced intercellular conductances that were volt age-gated at transjunctional potentials greater than or equal to 60 mV, and that were also closed by intracellular acidification. In contrast, Cx35 wa s unable to functionally interact with rodent connexins from the alpha or b eta subfamilies. Voltage-activated hemichannel currents were also observed in single oocytes expressing Cx35, and superfusing these oocytes with mediu m containing 100 mu M quinine resulted in a 1.8-fold increase in the magnit ude of the outward currents, but did not change the threshold of voltage ac tivation (membrane potential = +20 mV). Cx35 intercellular channels between paired oocytes were insensitive to quinine treatment. Both hemichannel act ivity and its modulation by quinine were seen previously in recordings from isolated skate horizontal cells. Voltage-activated currents of Cx46 hemich annels were also enhanced 1.6-fold following quinine treatment, whereas Cx4 3-injected oocytes showed no hemichannel activity in the presence, or absen ce, of quinine. Although the cellular localization of Cx35 is unknown, the functional characteristics of Cx35 in Xenopus oocytes are consistent with t he hemichannel and intercellular channel properties of skate horizontal cel ls.