Am J Physiol Cell Physiol AJP: Heart and Circulatory Physiology
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Am J Physiol Cell Physiol 283: C1627-C1636, 2002. First published August 7, 2002; doi:10.1152/ajpcell.00152.2001
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Vol. 283, Issue 6, C1627-C1636, December 2002

Regulation of volume-sensitive outwardly rectifying anion channels in pulmonary arterial smooth muscle cells by PKC

Juming Zhong1, Ge-Xin Wang1, William J. Hatton1, Ilia A. Yamboliev1, Michael P. Walsh2, and Joseph R. Hume1

1 Center of Biomedical Research Excellence, Department of Pharmacology, University of Nevada, Reno, Nevada 89557-0046; and 2 Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada T2N 4N1

We tested the possible role of endogenous protein kinase C (PKC) in the regulation of native volume-sensitive organic osmolyte and anion channels (VSOACs) in acutely dispersed canine pulmonary artery smooth muscle cells (PASMC). Hypotonic cell swelling activated native volume-regulated Cl- currents (ICl.vol) which could be reversed by exposure to phorbol 12,13-dibutyrate (0.1 µM) or by hypertonic cell shrinkage. Under isotonic conditions, calphostin C (0.1 µM) or Ro-31-8425 (0.1 µM), inhibitors of both conventional and novel PKC isozymes, significantly activated ICl.vol and prevented further modulation by subsequent hypotonic cell swelling. Bisindolylmaleimide (0.1 µM), a selective conventional PKC inhibitor, was without effect. Dialyzing acutely dispersed and cultured PASMC with epsilon V1-2 (10 µM), a translocation inhibitory peptide derived from the V1 region of epsilon PKC, activated ICl.vol under isotonic conditions and prevented further modulation by cell volume changes. Dialyzing PASMC with beta C2-2 (10 µM), a translocation inhibitory peptide derived from the C2 region of beta PKC, had no detectable effect. Immunohistochemistry in cultured canine PASMC verified that hypotonic cell swelling is accompanied by translocation of epsilon PKC from the vicinity of the membrane to cytoplasmic and perinuclear locations. These data suggest that membrane-bound epsilon PKC controls the activation state of native VSOACs in canine PASMC under isotonic and anisotonic conditions.

chloride channels; cell volume; protein kinase C


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