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J. Biol. Chem., Vol. 279, Issue 4, 2913-2921, January 23, 2004

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Molecular Cloning and Functional Expression of zfCx52.6

A NOVEL CONNEXIN WITH HEMICHANNEL-FORMING PROPERTIES EXPRESSED IN HORIZONTAL CELLS OF THE ZEBRAFISH RETINA^*

Georg Zoidl, Roberto Bruzzone, Svenja Weickert, Marian Kremer, Christiane Zoidl, Georgia Mitropoulou, Miduturu Srinivas¶, David C. Spray¶, and Rolf Dermietzel||

From the Department of Neuroanatomy and Molecular Brain Research, Ruhr-University-Bochum, University Street 150, D-44801 Bochum, Germany, the Department of Neuroscience, Institute Pasteur, 75015 Paris, France, the Department of Clinical Neurobiology, Interdisciplinary Center for Neurosciences (IZN), 69120 Heidelberg, Germany, and the ^¶Department of Neuroscience, Albert Einstein College of Medicine, Rose F. Kennedy Center, Bronx, New York 10461

Gap junction-mediated electrical coupling contributes to synchronous oscillatory activities of neurons, and considerable progress has been made in defining the molecular composition of gap junction channels. In particular, cloning and functional expression of gap junction proteins (connexins (Cx)) from zebrafish retina have shown that this part of the brain possesses a high degree of connexin diversity that may account for differential functional properties of electrical synapses. Here, we report the cloning and functional characterization of a new connexin, designated zebrafish Cx52.6 (zfCx52.6). This connexin shows little similarity to known connexins from fish and higher vertebrates. By combining in situ hybridization with Laser Capture Microdissection and RT-PCR, we found that this novel fish connexin is expressed in horizontal cells in the inner nuclear layer of the retina. Functional expression of zfCx52.6 in neuroblastoma cells and Xenopus oocytes led to functional gap junctional channels and, in single oocytes, induced large non-junctional membrane currents indicative of the formation of hemichannels, which were inhibited in reversible fashion by raising extracellular Ca²⁺ concentrations.

Received for publication, May 8, 2003 , and in revised form, October 28, 2003.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) GI:21311548.

^* This work was supported by the Deutsche Forschungsgemeinschaft, SFB 509, National Institutes of Health Grant MH65495, and the Association RETINA France. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed. Tel. 49-234-322-5003; Fax: 49-234-321-4655; E-mail: rolf.dermietzel{at}ruhr-unibochum.de.

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