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Kinetics of receptor-mediated endocytosis of albumin in cells derived from the proximal tubule of the kidney (opossum kidney cells): influence of Ca2+ and cAMP

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

In this study we investigated the effects of Ca2+ and cyclic adenosine monophosphate (cAMP) on the kinetic of receptor-mediated (RME) and fluidphase (FPE) endocytosis in opossum kidney (OK) cells, derived from the proximal tubule of the kidney. We used fluorescein isothiocyanate (FITC)-labelled albumin and FITC-labelled dextran as endocytotic substrates for RME and FPE, respectively. Removal of extracellular Ca2+ led to a dramatic decrease of the apparent affinity of RME, but did not influence the maximum endocytotic uptake rate (J max). Reduction of extracellular Ca2+ to 1 μmol/l had no effect. Apparent affinity of specific binding of albumin to the plasma membrane was increased to 200% of control in the absence of extracellular Ca2+, whereas maximum binding capacity was slightly decreased. FPE was not affected by removal of extracellular Ca2+. Additional removal of cytoplasmic Ca2+, using ionomycin, had no further effect on RME and did not affect FPE. Increases of cytoplasmic (using ionomycin at extracellular Ca2+ concentrations of 1 μmol/l or 1.2 mmol/l) or extracellular Ca2+ did not alter the kinetics of RME or FPE. Dibutyryl-cAMP reduced J max but left the apparent affinity of RME unchanged. FPE and albumin binding to the plasma membrane were not changed in the presence of cAMP. Removal of extracellular Ca2+ and addition of cAMP led to an alkalinization of endocytotic vesicles. Yet the alkalinization induced by removal of Ca2+ was significantly greater as compared to the alkalinization in the presence of cAMP. Endosomal alkalinization with bafilomycin A1 had no further effect in the absence of Ca2+, but reduced RME in the presence of cAMP. From these results we conclude that cAMP and extracellular Ca2+ influence the kinetics of RME in proximal tubular cells. The action of Ca2+ removal seems to be mediated partially by changes of endosomal pH. Ca2+ seems to be a prerequisite for RME but not a regulator, whereas cAMP may act as physiological inhibitor of RME.

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Authors and Affiliations

  1. Department of Physiology, University of Würzburg, Röntgenring 9, D-97070, Würzburg, Germany

    Ruth Freudinger & Stefen Silbernagl

  2. Department of Medicine, Royal North Shore Hospital, Sydney, Australia

    Michel Gekle & Sigrid Mildenberger

Authors
  1. Ruth Freudinger
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  2. Stefen Silbernagl
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  3. Michel Gekle
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  4. Sigrid Mildenberger
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Freudinger, R., Silbernagl, S., Gekle, M. et al. Kinetics of receptor-mediated endocytosis of albumin in cells derived from the proximal tubule of the kidney (opossum kidney cells): influence of Ca2+ and cAMP. Pflügers Arch 430, 374–380 (1995). https://doi.org/10.1007/BF00373912

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  • DOI: https://doi.org/10.1007/BF00373912

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