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A Variant of the Ca2+-Activated Cl Channel Best3 is Expressed in Mouse Exocrine Glands

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Abstract

Fluid secretion by exocrine glands requires the activation of an apical Ca2+-dependent Cl channel, the molecular identity of which is unknown. We found that mouse exocrine glands expressed an alternately spliced variant of Best3, a member of the Bestrophin (Vmd2) Ca2+-activated Cl channel gene family, whereas the heart expressed full-length Best3. The spliced transcript lacked exons 2, 3 and 6 (Best3-Δ2,3,6) and is predicted to generate an in-frame protein missing the entire cytoplasmic N terminus, the initial two transmembrane domains and part of the first intracellular loop. In addition to exocrine glands, the Best3-Δ2,3,6 splice variant transcript was detected in lung, testis and kidney. The parotid gland and heart expressed proteins of the predicted size for Best3-Δ2,3,6 and full-length Best3, respectively, that targeted to the plasma membrane in HEK293 cells. HEK293 cells expressing Best3 displayed Ca2+-dependent Cl− currents that were sensitive to the Cl channel blocker DIDS. In contrast, no Ca2+-dependent Cl− currents were detected in cells expressing Best3-Δ2,3,6. Cotransfection of Best3-Δ2,3,6 with Best3 or Best2 (also expressed in salivary gland acinar cells) had no significant effects on the currents generated by either of these Ca2+-dependent Cl channels. Our results demonstrate that exocrine glands express a unique splice variant of Best3. Nevertheless, Best3-Δ2,3,6 does not produce Ca2+-dependent Cl− currents, nor does it regulate the activity of Best2 or the full-length Best3 channel.

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Acknowledgements

We thank Laurie Koek, Mark Wagner and Jennifer Scantlin for technical assistance. We are also grateful to Dr. Ted Begenisich for discussions and critical reading of the manuscript. This work was supported in part by National Institutes of Health grants DE09692 and DE08921 (to J. E. M.).

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Authors and Affiliations

  1. Center for Oral Biology, Medical Center, University of Rochester School of Medicine and Dentistry, 601 Elmwood Ave., Box 611, Rochester, NY, 14642, USA

    Alaka Srivastava, Victor G. Romanenko, Mireya Gonzalez-Begne, Marcelo A. Catalán & James E. Melvin

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  1. Alaka Srivastava
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  2. Victor G. Romanenko
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  3. Mireya Gonzalez-Begne
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  4. Marcelo A. Catalán
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  5. James E. Melvin
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Correspondence to James E. Melvin.

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Alaka Srivastava, Victor G. Romanenko, and Mireya Gonzalez-Begne have contributed equally to this work.

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Srivastava, A., Romanenko, V.G., Gonzalez-Begne, M. et al. A Variant of the Ca2+-Activated Cl Channel Best3 is Expressed in Mouse Exocrine Glands. J Membrane Biol 222, 43–54 (2008). https://doi.org/10.1007/s00232-008-9098-4

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  • DOI: https://doi.org/10.1007/s00232-008-9098-4

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