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

Cellular & Molecular Biology Letters

Anna Wróbel²  Christer Lindqvist¹  Małgorzata Bobrowska-Hägerstrand¹  Henry Hägerstrand³ 

[1] Department of Biosciences, Åbo Akademi University, Åbo/Turku, Finland$$;Institute of Physics, Wrocław University of Technology, Wrocław, Poland$$;Department of Biosciences, Åbo Akademi University, Åbo/Turku, Finland$$Department of Biosciences, Cell Biology, Åbo Akademi University, Åbo/Turku, Finland$$

关键词: FM1-43;    Phospholipid scrambling;    Annexin V-FITC;    Phosphatidylserine exposure;    Human erythrocytes;    K562 cells;    Ionophores;    Amphiphiles;   

DOI  :  10.2478/s11658-014-0195-3

学科分类：分子生物学，细胞生物学和基因

来源: Uniwersytet Wroclawski * Wydzial Biotechnologii / University of Wroclaw, Faculty of Biotechnology

【 摘 要 】

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.

【 授权许可】

Unknown