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Aquaporin expression and cell volume regulation in the SV40 immortalized rat submandibular acinar cell line

  • Cell and Molecular Physiology
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Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

The amount of aquaporins present and the cellular ability to perform regulatory volume changes are likely to be important for fluid secretions from exocrine glands. In this work these phenomena were studied in an SV40 immortalized rat submandibular acinar cell line. The regulatory cell volume characteristics have not previously been determined in these cells. Cell volume regulation following hyposmotic exposure and aquaporin induction was examined with Coulter counter methodology, radioactive efflux studies, fura-2 fluorescence, and polymerase chain reaction and Western blot techniques. Cell volume regulation was inhibited by the K+ channel antagonists quinine and BaCl2 and the Cl− channel blocker 5-nitro-2-(3-phenypropylamino)benzoic acid. A concomitant increase in cellular 3H-taurine release and Ca2+ concentration was also observed. Chelation of both intra- and extracellular Ca2+ with EGTA and the Ca2+ ionophore A23187 did not, however, affect cell volume regulation. Aquaporin 5 (AQP5) mRNA and protein levels were upregulated in hyperosmotic conditions and downregulated upon return to isosmotic solutions, but were reduced by the mitogen-activated ERK-activating kinase (MEK) inhibitor U0126. A 24-h MEK inhibition also diminished hyposmotically induced cell swelling and cell volume regulation. In conclusion, it was determined that regulatory volume changes in this immortalized cell line are due to KCl and taurine efflux. In conditions that increased AQP5 levels, the cells showed a faster cell swelling and a more complete volume recovery following hyposmotic exposure. This response could be overturned by MEK inhibition.

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Acknowledgements

We are greatly in debt to Dr. David Quissell, Department of Craniofacial Biology, University of Colorado, for providing us with the acinar cell line and to Dr. Peter Agre, John Hopkins University School of Medicine, USA, for supplying the initial anti-rat AQP5 sera. We would also like to thank Dr. Kjell Fugelli, Department of Molecular Biosciences, University of Oslo, Norway, for assistance with the taurine efflux experiments. Excellent imaging service was provided by Hege Avsnes Dale, M.S., at the national technology platform Molecular Imaging Center at the University of Bergen, Norway, and supported by the functional genomics program (FUGE) in the Research Council of Norway.

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  1. Institute of Oral Biology, Faculty of Dentistry, University of Oslo, P.O. Box 1052, Blindern, 0316, Oslo, Norway

    Ann-Kristin Hansen & Hilde Kanli Galtung

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  1. Ann-Kristin Hansen
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  2. Hilde Kanli Galtung
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Correspondence to Hilde Kanli Galtung.

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Hansen, AK., Kanli Galtung, H. Aquaporin expression and cell volume regulation in the SV40 immortalized rat submandibular acinar cell line. Pflugers Arch - Eur J Physiol 453, 787–796 (2007). https://doi.org/10.1007/s00424-006-0158-2

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  • DOI: https://doi.org/10.1007/s00424-006-0158-2

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