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Role of F-actin in the activation of Na+-K+-Cl− cotransport by forskolin and vasopressin in mouse kidney cultured thick ascending limb cells

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Abstract

The influence of microtubules and F-actin on Na+-K+-Cl− cotransport was investigated in cultured cells derived from outer-medullary thick ascending limb tubules microdissected from the mouse kidney. The cultured cells contained Tamm-Horsfall protein, produced cAMP in response to dD-arginine vasopressin (dD-AVP), isoproterenol, prostaglandin E2 and forskolin (FK), and exhibited an ouabain-resistant furosemidesensitive (Or-Fs) component of 86Rb+ influx mediated by the Na+-K+-Cl− cotransporter. Both FK and dD-AVP stimulated the Or-Fs component of Rb+ influx. Neither agent altered the tubulin and cytokeratin networks nor the shape of the tight junction using a specific anti-ZO-1 antibody. In contrast, they did induce a marked redistribution of F-actin to the periphery of the cells delineating the tight junctions. Preincubation of the cells with nocodazole, to disrupt microtubules, did not alter the FK-or dD-AVP-elicited Or-Fs Rb+ influx. In contrast, phalloidin and NBD-phallicidin, which stabilize F-actin, markedly impaired the stimulation of Na+-K+-Cl− cotransport by FK or dD-AVP, without affecting the Na+-K+ ATPase pumps and the rate constant of 36Cl− and 86Rb+ efflux. These results strongly suggested that cAMP-stimulated Na+-K+-Cl− cotransport is linked to F-actin in renal TAL cells.

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Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale (INSERM), Unité 246, Institut Fédératif de Recherche Xavier Bichat, B.P. 416, 75870, 18, Paris Cedex, France

    M. S. Wu, M. Bens, F. Cluzeaud & A. Vandewalle

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  1. M. S. Wu
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  2. M. Bens
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  3. F. Cluzeaud
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  4. A. Vandewalle
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Additional information

This work was supported by INSERM. We thank Dr. James L. Madara for helpful discussions and for critically reading the manuscript. We also thank A. Loiseau (INSERM U251) for performing the kinetic analyses. We thank Mrs. G. Burger (LEICA GmbH, Heidelberg, Germany) for providing the photographs of CLSM analysis. Dr. Mai Szu Wu (Chang Gung Memorial Hospital, Taipei, Taiwan) holds an INSERM post-doctoral (Poste Vert) fellowship. We also thank Mrs. I. Gorne (ARKA laboratoire) for photographic works.

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Wu, M.S., Bens, M., Cluzeaud, F. et al. Role of F-actin in the activation of Na+-K+-Cl− cotransport by forskolin and vasopressin in mouse kidney cultured thick ascending limb cells. J. Membarin Biol. 142, 323–336 (1994). https://doi.org/10.1007/BF00233439

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  • DOI: https://doi.org/10.1007/BF00233439

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