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This Article Full Text Full Text (PDF) All Versions of this Article: 287/6/C1596    most recent 00225.2004v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (9) Citing Articles via Google Scholar Google Scholar Articles by Valiunas, V. Articles by White, T. W. Search for Related Content PubMed PubMed Citation Articles by Valiunas, V. Articles by White, T. W.

MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

Biophysical characterization of zebrafish connexin35 hemichannels

¹Department of Physiology and Biophysics, State University of New York, Stony Brook, New York 11794; and ²Department of Zoology, University of Manitoba, Winnipeg, Canada R3T 2N2

Submitted 6 May 2004 ; accepted in final form 23 July 2004

A subset of connexins can form unopposed hemichannels in expression systems, providing an opportunity for comparison of hemichannel gating properties with those of intact gap junction channels. Zebrafish connexin35 (Cx35) is a member of the Cx35/Cx36 subgroup of connexins highly expressed in the retina and brain. In the present study, we have shown that Cx35 expression in Xenopus oocytes and N2A cells produced large outward whole cell currents on cell depolarization. Using whole cell, cell-attached, and excised patch configurations, we obtained multichannel and single-channel current recordings attributable to the Cx35 hemichannels (I_hc) that were activated and increased by stepwise depolarization of membrane potential (V_m) and deactivated by hyperpolarization. The currents were not detected in untransfected N2A cells or in control oocytes injected with antisense Cx38. However, water-injected oocytes that were not treated with antisense showed activities attributable to Cx38 hemichannels that were easily distinguishable from Cx35 hemichannels by a significantly larger unitary conductance (_hc: 250–320 pS). The _hc of Cx35 hemichannels exhibited a pronounced V_m dependence; i.e., _hc increased/decreased with relative hyperpolarization/depolarization (_hc was 72 pS at V_m = –100 mV and 35 pS at V_m = 100 mV). Extrapolation to V_m = 0 mV predicted a _hc of 48 pS, suggesting a unitary conductance of intact Cx35 gap junction channels of 24 pS. Channel gating was also V_m dependent: open time declined with negative V_m and increased with positive V_m. The ability to break down the complex gating of intact intercellular channels into component hemichannels in vitro will help to evaluate putative physiological roles for hemichannels in vivo.

connexin; gating; retina

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