title: Amiloride-insensitive sodium channels are directly regulated by actin cytoskeleton dynamics in human lymphoma cells.
abstract: Sodium influx mediated by ion channels of plasma membrane underlies fundamental physiological processes in cells of blood origin. However, little is known about the single channel activity and regulatory mechanisms of sodium-specific channels in native cells. In the present work, we used different modes of patch clamp technique to examine ion channels involved in Na-transporting pathway in U937 human lymphoma cells. The activity of native non-voltage-gated sodium (NVGS) channels with unitary conductance of 10 pS was revealed in cell-attached, inside-out and whole-cell configurations. NVGS channel activity is directly controlled by submembranous actin cytoskeleton. Specifically, an activation of sodium channels in U937 cells in response to microfilament disassembly was demonstrated on single-channel and integral current level. Inside-out experiments showed that filament assembly on cytoplasmic membrane surface caused fast inactivation of the channels. Biophysical characteristics of NVGS channels in U937 cells were similar to that of epithelial sodium channels (ENaCs). However, we found that amiloride, a known inhibitor of DEG/ENaC, did not block NVGS channels in U937 cells. Whole-cell current measurements revealed no amiloride-sensitive component of membrane current. Our data show that cortical actin structures represent the main factor that controls the activity of amiloride-insensitive ENaC-like channels in human lymphoma cells.
authors: Institute of Cytology RAS, 194064 Tikhoretsky Ave. 4, St. Petersburg, Russia. Electronic address: anastasia.sudarikova@gmail.com.
authors: Anastasia V
authors: AV
authors: Sudarikova
authors: Institute of Cytology RAS, 194064 Tikhoretsky Ave. 4, St. Petersburg, Russia.
authors: Olga A
authors: OA
authors: Tsaplina
authors: Institute of Cytology RAS, 194064 Tikhoretsky Ave. 4, St. Petersburg, Russia.
authors: Vladislav I
authors: VI
authors: Chubinskiy-Nadezhdin
authors: Institute of Cytology RAS, 194064 Tikhoretsky Ave. 4, St. Petersburg, Russia.
authors: Elena A
authors: EA
authors: Morachevskaya
authors: Institute of Cytology RAS, 194064 Tikhoretsky Ave. 4, St. Petersburg, Russia; Department of Medical Physics, St. Petersburg State Polytechnical University, 29, Polytechnicheskaya St., 195251, St.Petersburg, Russia.
authors: Yuri A
authors: YA
authors: Negulyaev
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000584:Amiloride / Q000008:administration & dosage*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D062686:Epithelial Sodium Channel Blockers / Q000008:administration & dosage; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects*; D008223:Lymphoma / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D015222:Sodium Channels / Q000378:metabolism*
chemical_list: D062686:Epithelial Sodium Channel Blockers; D015222:Sodium Channels; D000584:Amiloride; D012964:Sodium
keywords: Actin cytoskeleton; Amiloride; Human lymphoma cell; Plasma membrane; Sodium channel