Functional properties of sodium channels in cholesterol-depleted K562 cells

• Published in: Cell and tissue biology Vol. 3; no. 5; pp. 461 - 469
• Main Authors: Sudarikova, A. V., Chubinsky-Nadezhdin, V. I., Negulyaev, Yu. A., Morachevskaya, E. A.
• Format: Journal Article
• Language: English
• Published: Dordrecht SP MAIK Nauka/Interperiodica 01.10.2009
• Subjects: Biomedical and Life Sciences; Cell Biology; Life Sciences; actin; methyl-beta-cyclodextrin; sodium channels; plasma membrane; cytoskeleton; cholesterol; cytochalasin; human leukemia cells
• ISSN: 1990-519X; 1990-5203
• DOI: 10.1134/S1990519X09050095

In the present paper, functional properties of nonvoltage-gated sodium channels in K562 cells were studied after cholesterol depletion, i.e., under conditions of the destruction of microdomains (rafts). For cholesterol depletion, cells were incubated with methyl-beta-cyclodextrin (MbCD), an oligosaccharide that selectively binds sterols. Single currents through sodium channels were recorded in cell-attached and inside-out experiments using the patch-clamp technique. After incubation with MbCD (2.5 or 5 mM), the activation of sodium channels in response to cytochalasin B or D was observed in both native cells and membrane fragments. Biophysical characteristics of sodium channels in cholesterol-depleted K562 cells were close to those in control; unitary conductance was 12 pS. Inside-out experiments with the use of globular actin have indicated that filament assembly on cytoplasmic membrane side causes an inactivation of sodium channels in the modified cells. These data imply that sodium channels in K562 cells are not associated with cholesterol-rich membrane microdomains. Possible mechanisms of the interaction of the plasma membrane and the cortical cytoskeleton are discussed.