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Accelerated suicidal erythrocyte death in Klotho-deficient mice

  • Molecular and Genomic Physiology
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Abstract

Klotho, a membrane protein mainly expressed in parathyroid glands, kidney, and choroid plexus, counteracts aging and increases the life span. Accordingly, life span is significantly shorter in Klotho-deficient mice (klotho −/−) than in their wild-type littermates (klotho +/+). The pleotropic effects of Klotho include inhibition of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) formation. Vitamin D-deficient diet reverses the shortening of life span in klotho −/− mice. In a variety of cells, 1,25(OH)2D3 stimulates Ca2+ entry. In erythrocytes, increased Ca2+ entry stimulates suicidal erythrocyte death, which is characterized by cell shrinkage and phosphatidylserine exposure at the erythrocyte surface. The present study explored the putative impact of Klotho on eryptosis. According to Fluo3 fluorescence, cytosolic Ca2+ concentration was significantly larger in klotho −/− erythrocytes as compared to klotho +/+ erythrocytes. According to annexin V-binding, phosphatidylserine exposure was significantly enhanced, and according to forward scatter, cell volume significantly decreased in klotho −/− erythrocytes as compared to klotho +/+ erythrocytes. Energy depletion (13 h glucose depletion) and oxidative stress (35 min 1 mM tert-butyl-hydroxyl-peroxide [tert-BOOH]) increased phosphatidylserine exposure to values again significantly larger in klotho −/− erythrocytes as compared to klotho +/+ erythrocytes. Reticulocyte number was significantly increased in klotho −/− mice, pointing to enhanced erythrocyte turnover. Vitamin D-deficient diet reversed the enhanced Ca2+ entry and annexin V-binding of klotho −/− erythrocytes. The present observations reveal a novel function of Klotho, i.e., the at least partially vitamin D-dependent regulation of cytosolic Ca2+ activity in and suicidal death of erythrocytes.

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Acknowledgments

The authors acknowledge the technical assistance of E. Faber and the meticulous preparation of the manuscript by Tanja Loch. This study was supported by the Deutsche Forschungsgemeinschaft, Nr. La 315/4-3 and La 315/6-1, and by the Promotionsverbund of the University of Tuebingen to S.K. M.F. was supported by a grant of the Carl-Zeiss-Stiftung.

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Authors and Affiliations

  1. Department of Physiology, University of Tübingen, Gmelinstr. 5, Tübingen, 72076, Germany

    Daniela S. Kempe, Teresa F. Ackermann, Stephanie S. Fischer, Saisudha Koka, Krishna M. Boini, Hasan Mahmud, Michael Föller & Florian Lang

  2. Department of Biochemistry and Molecular Biology and Department of Medicine, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555-1071, USA

    Kevin P. Rosenblatt

  3. Department of Pathology, The University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Blvd., Dallas, TX, 75390-9072, USA

    Makoto Kuro-o

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  1. Daniela S. Kempe
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  2. Teresa F. Ackermann
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Correspondence to Florian Lang.

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Kempe, D.S., Ackermann, T.F., Fischer, S.S. et al. Accelerated suicidal erythrocyte death in Klotho-deficient mice. Pflugers Arch - Eur J Physiol 458, 503–512 (2009). https://doi.org/10.1007/s00424-009-0636-4

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