Monitoring of membrane phospholipid scrambling in human erythrocytes and K562 cells with FM1-43 - a comparison with annexin V-FITC

• Autorzy: Wrobel A. , Bobrowska-Hagerstrand M. , Lindqvist C. , Hagerstrand H.
• Języki publikacji: EN

Abstrakty

EN

The styryl dye FM1-43 becomes highly fluorescent upon binding to cell membranes. The breakdown of membrane phospholipid asymmetry in ionophore-stimulated T-lymphocytes further increases this fluorescence [Zweifach, 2000]. In this study, the capacity of FM1-43 to monitor membrane phospholipid scrambling was explored using flow cytometry in human erythrocytes and human erythrocyte progenitor K562 cells. The Ca2+-dependent phosphatidylserine-specific probe annexin V-FITC was used for comparison. The presented data show that the loss of phospholipid asymmetry that could be induced in human erythrocytes by elevated intracellular Ca2+ or by structurally different membrane intercalated amphiphilic compounds increases the FM1-43 fluorescence two- to fivefold. The profile of FM1-43 fluorescence for various treatments resembles that of phosphatidylserine exposure reported by annexin V-FITC. FM1-43 detected the onset of scrambling more efficiently than annexin V-FITC. The amphiphile-induced scrambling was shown to be a Ca2+-independent process. Monitoring of scrambling in K562 cells caused by NEM-induced Ca2+-release from intracellular stores and by Ca2+ and ionophore A23187 treatment showed that the increase in FM1-43 fluorescence correlated well with the number of annexin V-FITC-detected phosphatidylserine-positive cells. The results presented here show the usefulness of FM1-43 as a Ca2+-independent marker of dissipation in asymmetric membrane phospholipid distribution induced by various stimuli in both nucleated and non-nucleated cells.

Słowa kluczowe

EN

monitoring; membrane phospholipid; man; erythrocyte; K562 cell; annexin; phospholipid scrambling; phosphatidylserine exposure; ionophore; amphiphile

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 2014
• Tom: 19
• Numer: 2
• Opis fizyczny: p.262-276,fig.,ref.

Twórcy

autor

Wrobel A.

• Institute of Physics, Wroclaw University of Technology, 50–370 Wroclaw, Poland

autor

Bobrowska-Hagerstrand M.

• Department of Biosciences, Abo Akademi University, Artillerigatan 6, FIN-20520 Abo/Turku, Finland

autor

Lindqvist C.

• Department of Biosciences, Abo Akademi University, Artillerigatan 6, FIN-20520 Abo/Turku, Finland

autor

Hagerstrand H.

• Department of Biosciences, Abo Akademi University, Artillerigatan 6, FIN-20520 Abo/Turku, Finland

Bibliografia

• 1. Bevers, E.M., Comfurius, P., Dekkers, D.W. and Zwaal, R.F. Lipid translocation across the plasma membrane of mammalian cells. Biochim. Biophys. Acta 1439 (1999) 317–330.
• 2. Daleke, D.L. Regulation of phospholipid asymmetry in the erythrocyte membrane. Curr. Opin. Hematol. 15 (2008) 191–195.
• 3. Zwaal, R.F., Comfurius, P. and Bevers, E.M. Surface exposure of phosphatidyl-serine in pathological cells. Cell. Mol. Life Sci. 62 (2005) 971–988.
• 4. Zhao, J., Zhou, Q., Wiedmer, T. and Sims, P.J. Level of expression of phospholipid scramblase regulates induced movement of phosphatidylserine to the cell surface. J. Biol. Chem. 273 (1998) 6603–6606.
• 5. Bassé, F., Stout, J.G., Sims, P.J. and Wiedmer, T. Isolation of an erythrocyte membrane protein that mediates Ca2+-dependent transbilayer movement of phospholipid. J. Biol. Chem. 271 (1996) 17205–17210.
• 6. Zhou, Q., Zhao, J., Stout, J.G., Luhm, R.A., Wiedmer, T. and Sims, P.J. Molecular cloning of human plasma membrane phospholipid scramblase. A protein mediating transbilayer movement of plasma membrane phospholipids. J. Biol. Chem. 272 (1997) 18240–18244.
• 7. Sahu, S.K., Gummadi, S.N., Manoj, N. and Aradhyam, G.K. Phospholipid scramblases: an overview. Arch. Biochem. Biophys. 462 (2007) 103–114.
• 8. Bevers, E.M. and Williamson, P.L. Phospholipid scramblase: An update. FEBS Lett. 584 (2010) 2724–2730.
• 9. Contreras, F.X., Sánchez-Magraner, L., Alonso, A. and Goñi, F.M. Transbilayer (flip-flop) lipid motion and lipid scrambling in membranes. FEBS Lett. 584 (2010) 1779–1786.
• 10. Zwaal, R.F. and Schroit, A.J. Pathophysiologic implications of membrane phospholipid asymmetry in blood cells. Blood 89 (1997) 1121–1132.
• 11. Williamson, P. and Schlegel, R.A. Transbilayer phospholipid movement and the clearance of apoptotic cells. Biochim. Biophys. Acta 1585 (2002) 53–63.
• 12. Dekkers, D.W., Comfurius, P., Bevers, E.M. and Zwaal, R.F. Comparison between Ca2+-induced scrambling of various fluorescently labeled lipid analogues in red blood cells. Biochem. J. 362 (2002) 741–747.
• 13. Williamson, P., Christie, A., Kohlin, T., Schlegel, R.A., Comfurius, P., Harmsma, M., Zwaal, R.F. and Bevers, E.M. Phospholipid scramblase activation pathways in lymphocytes. Biochemistry 40 (2001) 8065–8072.
• 14. Balasubramanian, K., Mirnikjoo, B. and Schroit, A.J. Regulated externalization of phosphatidylserine at the cell surface: implications for apoptosis. J. Biol. Chem. 282 (2007) 18357–18364.
• 15. Woon, L.A., Holland, J.W., Kable, E.P. and Roufogalis, B.D. Ca2+ sensitivity of phospholipid scrambling in human red cell ghosts. Cell Calcium 25 (1999) 313–320.
• 16. Wurth, G.A. and Zweifach, A. Evidence that cytosolic calcium increases are not sufficient to stimulate phospholipid scrambling in human T-lymphocytes. Biochem. J. 362 (2002) 701–708.
• 17. van Engeland, M., Kuijpers, H.J., Ramaekers, F.C., Reutelingsperger, C.P. and Schutte, B. Plasma membrane alterations and cytoskeletal changes in apoptosis. Exp. Cell. Res. 235 (1997) 421–430.
• 18. Fadeel, B., Gleiss, B., Högstrand, K., Chandra, J., Wiedmer, T., Sims, P.J., Henter, J.I., Orrenius, S. and Samali, A. Phosphatidylserine exposure during apoptosis is a cell-type-specific event and does not correlate with plasma membrane phospholipid scramblase expression. Biochem. Biophys. Res. Commun. 266 (1999) 504–511.
• 19. Bevers, E.M., Comfurius, P., van Rijn, J.L., Hemker, H.C. and Zwaal, R.F. Generation of prothrombin-converting activity and the exposure of phosphatidylserine at the outer surface of platelets. Eur. J. Biochem. 122 (1982) 429–436.
• 20. Bevers, E.M., Comfurius, P. and Zwaal, R.F. Changes in membrane phospholipid distribution during platelet activation. Biochim. Biophys. Acta 736 (1983) 57–66.
• 21. Blumenfeld, N., Zachowski, A., Galacteros, F., Beuzard, Y. and Devaux, P.F. Transmembrane mobility of phospholipids in sickle erythrocytes: effect of deoxygenation on diffusion and asymmetry. Blood 77 (1991) 849–854.
• 22. Utsugi, T., Schroit, A.J., Connor, J., Bucana, C.D. and Fidler, I.J. Elevated expression of phosphatidylserine in the outer membrane leaflet of human tumor cells and recognition by activated human blood monocytes. Cancer Res. 51 (1991) 3062–3066.
• 23. Schroit, A.J., Madsen, J.W. and Tanaka, Y. In vivo recognition and clearance of red blood cells containing phosphatidylserine in their plasma membranes. J. Biol. Chem. 260 (1985) 5131–5138.
• 24. Connor, J., Pak, C.C. and Schroit, A.J. Exposure of phosphatidylserine in the outer leaflet of human red blood cells. Relationship to cell density, cell age, and clearance by mononuclear cells. J. Biol. Chem. 269 (1994) 2399–2404.
• 25. Fadok, V.A., Voelker, D.R., Campbell, P.A., Cohen, J.J., Bratton, D.L. and Henson, P.M. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J. Immunol. 148 (1992) 2207–2216.
• 26. Martin, S.J., Reutelingsperger, C.P., McGahon, A.J., Rader, J.A., van Schie, R.C., LaFace, D.M. and Green, D.R. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J. Exp. Med. 182 (1995) 1545–1556.
• 27. Kuypers, F.A., Lewis, R.A., Hua, M., Schott, M.A., Discher, D., Ernst, J.D. and Lubin, B.H. Detection of altered membrane phospholipid asymmetry in subpopulations of human red blood cells using fluorescently labeled annexin V. Blood 87 (1996) 1179–1187.
• 28. Wood, B.L., Gibson, D.F. and Tait, J.F. Increased erythrocyte phosphatidylserine exposure in sickle cell disease: flow-cytometric measurement and clinical associations. Blood 88 (1996) 1873–1880.
• 29. Trotter, P.J., Orchard, M.A. and Walker, J.H. Ca2+ concentration during binding determines the manner in which annexin V binds to membranes. Biochem. J. 308 (1995) 591–598.
• 30. Koopman, G., Reutelingsperger, C.P., Kuijten, G.A., Keehnen, R.M., Pals, S.T. and van Oers, M.H. Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84 (1994) 1415–1420.
• 31. Kamp, D., Sieberg, T. and Haest, C.W. Inhibition and stimulation of phospholipid scrambling activity. Consequences for lipid asymmetry, echinocytosis, and microvesiculation of erythrocytes. Biochemistry 40 (2001) 9438–9446.
• 32. Hanshaw, R.G. and Smith, B.D. New reagents for phosphatidylserine recognition and detection of apoptosis. Bioorg. Med. Chem. 13 (2005) 5035–5042.
• 33. Zweifach, A. FM1-43 reports plasma membrane phospholipid scrambling in T-lymphocytes. Biochem. J. 349 (2000) 255–260.
• 34. Cochilla, A.J., Angleson, J.K. and Betz, W.J. Monitoring secretory membrane with FM1-43 fluorescence. Annu. Rev. Neurosci. 22 (1999) 1–10.
• 35. Betz, W.J., Mao, F. and Smith, C.B. Imaging exocytosis and endocytosis. Curr. Opin. Neurobiol. 6 (1996) 365–371.
• 36. Schote, U. and Seelig, J. Interaction of the neuronal marker dye FM1-43 with lipid membranes. Thermodynamics and lipid ordering. Biochim. Biophys. Acta 1415 (1998) 135–146.
• 37. Hägerstrand, H., Holmström, T.H., Bobrowska-Hägerstrand, M., Eriksson, J.E. and Isomaa, B. Amphiphile-induced phosphatidylserine exposure in human erythrocytes. Mol. Membr. Biol. 15 (1998) 89–95.
• 38. Lozzio, C.B. and Lozzio, B.B. Human chronic myelogenous leukemia cellline with positive Philadelphia chromosome. Blood 45 (1975) 321–334.
• 39. Isomaa, B., Hägerstrand, H. and Paatero, G. Shape transformations induced by amphiphiles in erythrocytes. Biochim. Biophys. Acta 899 (1987) 93–103.
• 40. Williamson, P., Mattocks, K. and Schlegel, R.A. Merocyanine 540, a fluorescent probe sensitive to lipid packing. Biochim. Biophys. Acta 732 (1983) 387–393.
• 41. Wu, Y., Yeh, F.L., Mao, F. and Chapman, E.R. Biophysical characterization of styryl dye-membrane interactions. Biophys. J. 97 (2009) 101–109.