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Thermal inactivation of volume-sensitive K+,Cl− cotransport and plasma membrane relief changes in human erythrocytes

  • Ion Channels, Receptors and Transporters
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Abstract

Previously, we reported that in mammalian erythrocytes irreversible annealing of spectrin heterodimers at 49–50 °C abolished cell volume-dependent regulation of ion carriers, thus suggesting an implication of a two-dimensional (2D) membrane carcass in volume sensing and/or signal transduction. To further examine this hypothesis, we employed atomic force microscopy. This method revealed folded membrane relief of fixed human erythrocytes with an average wave height of 3–5 nm covered by globular structures with a diameter of 40–50 nm and an average height of 1–2 nm. Erythrocyte swelling caused by reduction of medium osmolality decreased the height of membrane surface waves by 40 % and increased K+,Cl− cotransport by approximately sixfold. Both volume-sensitive changes of membrane relief and activity of K+,Cl− cotransporter were abolished by a 10-min preincubation at 50 °C. Our results strongly suggest that volume-dependent alterations of the human erythrocyte membrane relief are caused by reorganization of the 2D spectrin–actin network contributing to regulation of the activity of volume-sensitive ion transporters.

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References

  1. Adragna N, Di Fulvio M, Lauf PK (2004) Regulation of K-Cl cotransport: from function to genes. J Membr Biol 201:109–137

    Article  PubMed  CAS  Google Scholar 

  2. Anfinogenova YJ, Rodriguez X, Grygorczyk R, Adragna N, Lauf PK, Hamet P, Orlov SN (2001) Swelling-induced K+ fluxes in vascular smooth muscle cells are mediated by charybdotoxin-sensitive K+ channels. Cell Physiol Biochem 11:295–310

    Article  PubMed  CAS  Google Scholar 

  3. Brandts JF, Taverna RD, Salasivan E, Lysko KA (1978) Calorimetric studies of structural transitions of human erythrocyte membrane. Studies of B and C transitions. Biochim Biophys Acta 512:566–578

    Article  PubMed  CAS  Google Scholar 

  4. Burton NM, Bruce LJ (2011) Modeling the structure of the red cell membrane. Biochem Cell Biol 89:200–215

    Article  PubMed  CAS  Google Scholar 

  5. Cheng Y, Liu M, Li R, Wang C, Bai C, Wang K (1999) Gadolinium induces domain and pore formation of human erythrocyte membrane: an atomic force microscopic study. Biochim Biophys Acta 1421:249–260

    Article  PubMed  CAS  Google Scholar 

  6. Evans J, Gratzer W, Mohandas N, Parker K, Sleep J (2008) Fluctuations of the red blood cell membrane: relation to mechanical properties and lack of ATP-dependence. Biophys J 94:4134–4144

    Article  PubMed  CAS  Google Scholar 

  7. Francis LW, Lewis PD, Wright CJ, Conlan RS (2010) Atomic force microscopy comes of age. Biol Cell 102:133–143

    CAS  Google Scholar 

  8. Garay R, Nazaret C, Hannaert R, Cragoe E (1988) Demonstration of a K+, Cl− cotransport system in human red blood cells by its sensitivity to [(dihydroindenyl)oxy]alcanoic acid regulation of cell swelling and distinction from bumetanide-sensitive Na+, K+, Cl− cotransport system. Mol Pharmacol 33:696–701

    PubMed  CAS  Google Scholar 

  9. Girasole M, Pompeo G, Cricenti A, Congiu-Castellano A, Andreola F, Serafino A, Frazer BH, Boumis G, Amiconi G (2007) Roughness of the plasma membrane as an independent morphological parameter to study RBCs: A quantitative atomic force microscopy investigation. Biochim Biophys Acta 1768:1268–1276

    Article  PubMed  CAS  Google Scholar 

  10. Hoffmann EK, Lambert IH, Pedersen SF (2009) Physiology of cell volume regulation in vertebrates. Physiol Rev 89:193–277

    Article  PubMed  CAS  Google Scholar 

  11. Janmey PA, Lindberg U (2004) Cytoskeletal regulation: rich in lipids. Nat Rev Mol Cell Biol 5:658–666

    Article  PubMed  CAS  Google Scholar 

  12. Lang F, Busch G, Ritter M, Volkl H, Waldegger S, Gulbins E, Haussinger D (1998) Functional significance of cell volume regulatory mechanisms. Physiol Rev 78:247–306

    PubMed  CAS  Google Scholar 

  13. Lauf PK, Adragna NC (2000) K-Cl cotransport: properties and molecular mechanism. Cell Physiol Biochem 10:341–354

    Article  PubMed  CAS  Google Scholar 

  14. Mongin AA, Orlov SN (2001) Mechanisms of cell volume regulation and possible nature of the cell volume sensor. Pathophysiology 8:77–88

    Article  PubMed  CAS  Google Scholar 

  15. Nasuhoglu C, Feng S, Mao Y, Shammat I, Yamamato M, Earnest S, Lemmon M, Hilgemann DW (2002) Modulation of cardiac PIP2 by cardioactive hormones and other physiologically relevant interventions. Am J Physiol Cell Physiol 283:C223–C234

    Article  PubMed  CAS  Google Scholar 

  16. Nielsen DK, Jensen AK, Harbak H, Christensen SC, Simonsen LO (2007) Cell content of phosphatidylinositol (4,5)biphosphate in Ehrlich mouse ascites tumour cells in response to cell volume perturbations in anisotonic and isosmotic media. J Physiol 582:1027–1036

    Article  PubMed  CAS  Google Scholar 

  17. Oberleithner H (2008) Nanophysiology: fact or fiction? Pfluger Arch.- Eur. J Physiol 456:1–2

    CAS  Google Scholar 

  18. Ohta Y, Otsuka C, Okamoto H (2003) Changes in surface roughness of erythrocytes due to shear stress: atomic force microscopic visualization of the surface microstructure. J Artif Organs 6:101–105

    PubMed  Google Scholar 

  19. O’Neill WC (1999) Physiological significance of volume-regulated transporters. Am J Physiol 276:C995–C1011

    PubMed  Google Scholar 

  20. Orlov SN, Kolosova IA, Cragoe EJ, Gurlo TG, Mongin AA, Aksentsev SL, Konev SV (1993) Kinetics and peculiarities of thermal inactivation of volume-dependent Na/H exchange, Na, K,2Cl cotransport and K, Cl cotransport in rat erythrocytes. Biochim Biophys Acta 1151:186–192

    Article  PubMed  CAS  Google Scholar 

  21. Orlov SN, Kuznetsov SR, Kolosova IA, Aksentsev SL, Konev SV (1997) Volume-dependent regulation of ion transporters in human and rat erythrocytes: role of cytoskeleton and protein phosphorylation. Russ Physiol J 83(No5–6):119–147

    CAS  Google Scholar 

  22. Orlov SN, Pokudin NI, Kotelevtsev YV, Gulak PV (1989) Volume-dependent regulation of ion transport and membrane phosphorylation in human and rat erythrocytes. J Membr Biol 107:105–117

    Article  PubMed  CAS  Google Scholar 

  23. Park Y, Best CA, Auth T, Gov NS, Safran SA, Popescu G, Suresh S, Feld MS (2010) Metabolic remodelling of the human red blood cell membrane. Proc Natl Acad Sci USA 107:1289–1294

    Article  PubMed  CAS  Google Scholar 

  24. Repin NV, Bobrova EN, Repina SV (2008) Thermally induced transformation of mammalian red blood cells during hyperthermia. Bioelectrochemistry 73:101–105

    Article  PubMed  CAS  Google Scholar 

  25. Sachs JR (1998) How do red blood cells know how big they are? In: Okada Y (ed) Cell volume regulation: the molecular mechanism and volume sensing machinery. Elsevier Science, Tokyo, pp 3–13

    Google Scholar 

  26. Shnyrov VL, Orlov SN, Zhadan GG, Pokudin NI (1990) Thermal inactivation of membrane proteins, volume-dependent Na+, K+ cotransport, and protein kinase C activator-induced changes of the shape of human and rat erythrocytes. Biomed Biochim Acta 49:445–453

    PubMed  CAS  Google Scholar 

  27. Strange K (2004) Cellular volume homeostasis. Adv Physiol Educ 28:155–159

    Article  PubMed  Google Scholar 

  28. Tuvia S, Levin S, Bitler A, Korenstein R (1998) Mechanical fluctuation of the membrane-skeleton are dependent on F-actin ATPase in human erythrocytes. J Cell Biol 141:1551–1561

    Article  PubMed  CAS  Google Scholar 

  29. Yamamoto M, Chen MZ, Wang YJ, Sun HQ, Wei Y, Martinez M, Yin HL (2006) Hypertonic stress increases phosphatidylinositol 4,5-biphosphate levels by activating PIP5KIbeta. J Biol Chem 281:32630–32638

    Article  PubMed  CAS  Google Scholar 

  30. Yusipovich AI, Zagubizhenko MV, Levin GG, Platonova A, Parshina EY, Grygorczyk R, Maksimov GV, Rubin AB, Orlov SN (2011) Laser interference microscopy of amphibian erythrocytes: impact of cell volume and refractive index. J Microsc 244:223–229

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by grants from the Natural Sciences and Engineering Research Council of Canada (to R.G. and S.N.O.), the Russian Foundation for Fundamental Research (12-04-00033-a to S.N.O; 10-04-00835-а to G.V.M. and E.Y.P.), and the Ministry of Education and Science of the Russian Federation (projects ##8477 and 8162). Manuscript editing by Ovid Da Silva and logistical services from the Research Support Office, CRCHUM are appreciated.

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Authors and Affiliations

  1. Department of Biophysics and Laboratory of Physical Chemistry of Biomembranes, Faculty of Biology, M.V. Lomonosov Moscow State University, Moscow, Russia

    E. Yu. Parshina, A. I. Yusipovich, A. A. Platonova, G. V. Maksimov & S. N. Orlov

  2. Department of Medicine and Centre de recherche, Centre hospitalier de l’Université de Montréal (CRCHUM), Technopôle Angus, 2901 Rachel Est, Montreal, QC, H1W 4A4, Canada

    A. A. Platonova, R. Grygorczyk & S. N. Orlov

  3. Institute of General Pathology and Pathophysiology, Russian Academy of Medical Sciences, Moscow, Russia

    S. N. Orlov

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  1. E. Yu. Parshina
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  2. A. I. Yusipovich
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  3. A. A. Platonova
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  4. R. Grygorczyk
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  5. G. V. Maksimov
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  6. S. N. Orlov
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Correspondence to S. N. Orlov.

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Parshina, E.Y., Yusipovich, A.I., Platonova, A.A. et al. Thermal inactivation of volume-sensitive K+,Cl− cotransport and plasma membrane relief changes in human erythrocytes. Pflugers Arch - Eur J Physiol 465, 977–983 (2013). https://doi.org/10.1007/s00424-013-1221-4

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  • DOI: https://doi.org/10.1007/s00424-013-1221-4

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