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Ca2+- and H+-dependent effects of crude bacterial phospholipase C on the hydroosmotic response of toad urinary bladder to serosal hypertonicity

  • Transport Processes, Metabolism and Endocrinology; Crinology; Kidney, Gastrointestinal Tract, and Exocrine Glands
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Abstract

Phospholipase C (EC 3.1.4.3.) from Clostridium perfringens (crude extracts) was used to study the role of phospholipids in the osmotic permeability of the urinary bladder of the toad. When added to the serosal bath (430 mU/ml) it inhibited the effects of antidiurectic hormone (ADH) and exogenous cyclic AMP. Under the same conditions the increase in osmotic flow produced by serosal hypertonicity (SH) was slightly enhanced by the lipase. The hydroosmotic effect of SH was greatly potentiated by the lipase by decreasing 10-fold the Ca2+ concentration. The SH-induced flow was inhibited by the lipase if the Ca2+ or the H+ concentration was increased 10-fold, but not if the increase in positive charges was produced by a concentration of Mg2+. Phospholipase C had no effect on the action of either ADH or SH if added to the mucosal bath. Serosal neuraminidase or phospholipase A2 could not mimic the effect of phospholipase C on SH. The effect of phospholipase C on the response to SH was not modified if fatty acid-free bovine serum albumin was added to the bath. Therefore, the release of products of lipolysis into the bath do not seem to be responsible for the effects of phospholipase C on SH-induced water flow. The results suggest that the effects of the enzyme on the composition and rearrangement of lipids at the basolateral membrane produce modifications of the water flow. Ca2+ and H+ may modify the enzyme-substrate interaction, suggesting that different phospholipids may be differentially involved in the control of water permeability of the basolateral membrane. Changes in Ca2+ and H+ concentration may also activate or suppress different lipolytic enzymes of the nonhomogenous enzymatic complex employed. These results support the idea that ADH and SH converge in a final common site of action in order to increase the water permeability of the apical membrane, but they do so by activating different steps, SH acting mainly at a post-cyclic AMP site.

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Author notes

  1. Marcos A. Hardy

    Present address: Center for Polypeptide and Membrane Research, The Mount Sinai Medical Center of the City University of New York, Annenberg Building 21-86, One Gustave L. Levy Place, 10029, New York, NY, USA

Authors and Affiliations

  1. Department of Medicine (Division of Nephrology) and Pharmacology, University of Miami School of Medicine, 33101, Miami, Florida, USA

    Marcos A. Hardy

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  1. Marcos A. Hardy
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Hardy, M.A. Ca2+- and H+-dependent effects of crude bacterial phospholipase C on the hydroosmotic response of toad urinary bladder to serosal hypertonicity. Pflugers Arch. 402, 171–175 (1984). https://doi.org/10.1007/BF00583331

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  • DOI: https://doi.org/10.1007/BF00583331

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