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J. Biol. Chem., Vol. 279, Issue 35, 36993-37003, August 27, 2004

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Connexin 48.5 Is Required for Normal Cardiovascular Function and Lens Development in Zebrafish Embryos^*

Shaohong Cheng, Teresa Shakespeare, Rickie Mui, Thomas W. White, and Gunnar Valdimarsson¶||

From the Department of Zoology and ^¶Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada and Department of Physiology and Biophysics, State University of New York, Stony Brook, New York 11794

Gap junctions are composed of connexin (Cx) proteins and mediate intercellular communication required for many developmental and physiological processes. Here we describe the isolation and characterization of Cx48.5, a zebrafish connexin with the highest sequence identity to mammalian Cx46. Expression analysis showed that Cx48.5 is expressed in the adult and embryonic lens and heart, adult testis, and transiently in the embryonic otic vesicles. Injection of Cx48.5 cRNA into Xenopus oocytes elicited intercellular electrical coupling with voltage sensitivity similar to mammalian Cx46. In single oocytes, Cx48.5 also induced large outward currents on depolarization, consistent with gap-junctional hemichannels. Disruption of Cx48.5 expression in embryos with antisense morpholino oligos (morpholinos) revealed that Cx48.5 has an essential role in the maintenance of lens homeostasis. The morpholino-treated embryos also developed small lenses and eyes as well as severe cardiovascular abnormalities.

Received for publication, February 6, 2004 , and in revised form, June 3, 2004.

^* This work was supported by research grants (to G. V.) and scholarships (to S. C.) from the Natural Sciences and Engineering Research Council of Canada and the Manitoba Health Research Council and by National Institutes of Health Grants EY13163 and DC06652 (to T. W. W.). 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.

The on-line version of this article (available at http://www.jbc.org) contains supplemental videos.

^|| To whom correspondence should be addressed: Dept. of Zoology, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada. Tel.: 204-4746170; Fax: 204-4747588; E-mail: valdimar{at}cc.umanitoba.ca.

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