title: ÐÑÑÐ²Ð»ÐµÐ½Ð¸Ðµ Î´-ÑÑÐ±ÑÐµÐ´Ð¸Ð½Ð¸ÑÑ ENaC Ð² ÑÐ¾ÑÑÐ°Ð²Ðµ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾-Ð°ÐºÑÐ¸Ð²Ð½Ð¾Ð³Ð¾ Ð½Ð°ÑÑÐ¸ÐµÐ²Ð¾Ð³Ð¾ ÐºÐ°Ð½Ð°Ð»Ð° Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Ð¼Ð¸ÐµÐ»Ð¾Ð¸Ð´Ð½Ð¾Ð¹ Ð»ÐµÐ¹ÐºÐµÐ¼Ð¸Ð¸ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ° Ð562
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: The detection of the ENaC Î´ subunit in the composition of functionally active sodium channels in K562 cells
Creators: name: ÐÑÑÐ¸ÐºÐ¾Ð²Ð°, ÐÐ°ÑÑÑ
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2020
Subjects: Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»-Ð½ÐµÐ·Ð°Ð²Ð¸ÑÐ¸Ð¼ÑÐµ Ð½Ð°ÑÑÐ¸ÐµÐ²ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ
Ð°ÐºÑÐ¸Ð½-ÑÐ¿ÑÐ°Ð²Ð»ÑÐµÐ¼ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ
ÑÐ¿Ð¸ÑÐµÐ»Ð¸Ð°Ð»ÑÐ½ÑÐµ Ð½Ð°ÑÑÐ¸ÐµÐ²ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ ENaC
Î´-ÑÑÐ±ÑÐµÐ´Ð¸Ð½Ð¸ÑÐ° ENaC
ÐºÐ°Ð¿ÑÐ°Ð·ÐµÐ¿Ð¸Ð½
ÐºÐ»ÐµÑÐºÐ¸ Ð»ÐµÐ¹ÐºÐµÐ¼Ð¸Ð¸ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ°
patch-clamp
voltage-independent sodium channels
epithelial sodium channels ENaC
actin-gated channels
Î´-ENaC subunit
capsazepine
human leukemia cells

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¸Ð·ÑÑÐµÐ½Ð¸Ñ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»-Ð½ÐµÐ·Ð°Ð²Ð¸ÑÐ¸Ð¼ÑÑ Ð°ÐºÑÐ¸Ð½-ÑÐ¿ÑÐ°Ð²Ð»ÑÐµÐ¼ÑÑ Ð½Ð°ÑÑÐ¸ÐµÐ²ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Ð»ÐµÐ¹ÐºÐµÐ¼Ð¸Ð¸ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ° Ð562. ÐÐ¾ ÑÐ²Ð¾Ð¸Ð¼ ÑÐ°ÑÐ°ÐºÑÐµÑÐ¸ÑÑÐ¸ÐºÐ°Ð¼ Ð¸ Ð¼ÐµÑÐ°Ð½Ð¸Ð·Ð¼Ð°Ð¼ ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ Ð¸ÑÑÐ»ÐµÐ´ÑÐµÐ¼ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ Ð±Ð»Ð¸Ð·ÐºÐ¸ Ðº ÑÐµÐ¼ÐµÐ¹ÑÑÐ²Ñ ENaC, Ð·Ð° Ð¸ÑÐºÐ»ÑÑÐµÐ½Ð¸ÐµÐ¼ ÑÑÐ²ÑÑÐ²Ð¸ÑÐµÐ»ÑÐ½Ð¾ÑÑÐ¸ Ðº Ð¸Ð½Ð³Ð¸Ð±Ð¸ÑÐ¾ÑÑ Ð°Ð¼Ð¸Ð»Ð¾ÑÐ¸Ð´Ñ. ÐÑÑÐ°ÐµÑÑÑ Ð¾ÑÐºÑÑÑÑÐ¼ Ð²Ð¾Ð¿ÑÐ¾Ñ Ð¾Ð± Ð¸Ñ Ð²Ð¾Ð·Ð¼Ð¾Ð¶Ð½Ð¾Ð¼ ÑÑÐ±ÑÐµÐ´Ð¸Ð½Ð¸ÑÐ½Ð¾Ð¼ ÑÐ¾ÑÑÐ°Ð²Ðµ. Ð¦ÐµÐ»ÑÑ ÑÐ°Ð±Ð¾ÑÑ ÑÐ²Ð»ÑÐ»Ð¾ÑÑ Ð¸Ð·ÑÑÐµÐ½Ð¸Ðµ Ð²Ð¾Ð·Ð¼Ð¾Ð¶Ð½Ð¾Ð³Ð¾ ÑÑÐ°ÑÑÐ¸Ñ Î´-ÑÑÐ±ÑÐµÐ´Ð¸Ð½Ð¸ÑÑ ENaC Ð² ÑÐ¾ÑÑÐ°Ð²Ðµ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾-Ð°ÐºÑÐ¸Ð²Ð½Ð¾Ð³Ð¾ Ð½Ð°ÑÑÐ¸ÐµÐ²Ð¾Ð³Ð¾ ÐºÐ°Ð½Ð°Ð»Ð° Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Ð562.
Ð ÐºÐ¾Ð½ÑÐ¸Ð³ÑÑÐ°ÑÐ¸Ð¸ whole-cell Ð¼ÐµÑÐ¾Ð´Ð° Ð¿Ð°ÑÑ-ÐºÐ»Ð°Ð¼Ð¿ Ð·Ð°ÑÐµÐ³Ð¸ÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð»Ð¸ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ð¾Ð´Ð¸Ð½Ð¾ÑÐ½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð², Ð°ÐºÑÐ¸Ð²Ð¸ÑÑÐµÐ¼ÑÑ Ð¿ÑÐ¸ ÑÐ°Ð·Ð±Ð¾ÑÐºÐµ ÑÐ¸ÑÐ¾ÑÐºÐµÐ»ÐµÑÐ° Ð¦Ð¸ÑÐ¾ÑÐ°Ð»Ð°Ð·Ð¸Ð½Ð¾Ð¼ Ð (100% Ð¾Ð¿ÑÑÐ¾Ð²), Ð¸Ð·ÑÑÐ¸Ð»Ð¸ Ð¸Ñ ÑÐ°ÑÐ°ÐºÑÐµÑÐ¸ÑÑÐ¸ÐºÐ¸ Ð² Na+ Ð¸Ð»Ð¸ Li+ ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐµÐ¼ Ð²Ð½ÐµÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¼ ÑÐ°ÑÑÐ²Ð¾ÑÐµ. ÐÑÐ¾Ð²Ð¾Ð´Ð¸Ð¼Ð¾ÑÑÑ ÑÐ°ÑÑÐ¸ ÑÐµÐ³Ð¸ÑÑÑÐ¸ÑÑÐµÐ¼ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð´Ð»Ñ Li+ Ð±ÑÐ»Ð° ÑÐ½Ð¸Ð¶ÐµÐ½Ð° Ð¿Ð¾ ÑÑÐ°Ð²Ð½ÐµÐ½Ð¸Ñ Ñ Na+ (145 Ð¼Ð). Ð­ÑÐ° Ð¾ÑÐ¾Ð±ÐµÐ½Ð½Ð¾ÑÑÑ ÑÐ²Ð»ÑÐµÑÑÑ Ð¾ÑÐ½Ð¾Ð²Ð½ÑÐ¼ Ð±Ð¸Ð¾ÑÐ¸Ð·Ð¸ÑÐµÑÐºÐ¸Ð¼ Ð¿ÑÐ¸Ð·Ð½Ð°ÐºÐ¾Ð¼ ENaC, ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ¸Ñ Î´ ÑÑÐ±ÑÐµÐ´Ð¸Ð½Ð¸ÑÑ (Ð²Ð¼ÐµÑÑÐ¾ Î±). Ð ÐµÐ·ÑÐ»ÑÑÐ°ÑÑ Ð¸Ð¼Ð¼ÑÐ½Ð¾ÑÐ»ÑÐ¾ÑÐµÑÑÐµÐ½ÑÐ½Ð¾Ð³Ð¾ Ð¾ÐºÑÐ°ÑÐ¸Ð²Ð°Ð½Ð¸Ñ Ð¿Ð¾Ð´ÑÐ²ÐµÑÐ´Ð¸Ð»Ð¸ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ñ Î´-hENaC (Ð½Ð°ÑÑÐ´Ñ Ñ Î±-, Î²-, Î³-ENaC).
Ð Ð¾Ð¿ÑÑÐ°Ñ whole-cell Ð¾Ð±Ð½Ð°ÑÑÐ¶ÐµÐ½Ð¾, ÑÑÐ¾ Ð²Ð½ÐµÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ðµ Ð´Ð¾Ð±Ð°Ð²Ð»ÐµÐ½Ð¸Ðµ ÐºÐ°Ð¿ÑÐ°Ð·ÐµÐ¿Ð¸Ð½Ð°, Ð°ÐºÑÐ¸Ð²Ð°ÑÐ¾ÑÐ° Î´-ENaC, Ð²ÑÐ·ÑÐ²Ð°Ð»Ð¾ Ð°ÐºÑÐ¸Ð²Ð°ÑÐ¸Ñ Ð½Ð°ÑÑÐ¸ÐµÐ²ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð² 67% Ð¾Ð¿ÑÑÐ¾Ð². ÐÐ¸Ð¾ÑÐ¸Ð·Ð¸ÑÐµÑÐºÐ¸Ðµ ÑÐ°ÑÐ°ÐºÑÐµÑÐ¸ÑÑÐ¸ÐºÐ¸ ÐºÐ°Ð½Ð°Ð»Ð¾Ð², Ð°ÐºÑÐ¸Ð²Ð¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ ÐºÐ°Ð¿ÑÐ°Ð·ÐµÐ¿Ð¸Ð½Ð¾Ð¼, ÑÐ¾Ð²Ð¿Ð°Ð´Ð°Ð»Ð¸ Ñ ÑÐ°ÑÐ°ÐºÑÐµÑÐ¸ÑÑÐ¸ÐºÐ°Ð¼Ð¸ Ð°ÐºÑÐ¸Ð½-ÑÐ¿ÑÐ°Ð²Ð»ÑÐµÐ¼ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð². ÐÑÐ¸ Ð·Ð°Ð¼ÐµÐ½Ðµ Na+ Ð² ÐºÐ°Ð¼ÐµÑÐµ Ð½Ð° Li+ Ð½Ð°Ð±Ð»ÑÐ´Ð°Ð»Ð¸ ÑÐ¼ÐµÐ½ÑÑÐµÐ½Ð¸Ðµ Ð°Ð¼Ð¿Ð»Ð¸ÑÑÐ´Ñ ÑÐ¾ÐºÐ¾Ð² Ð¸ Ð¿ÑÐ¾Ð²Ð¾Ð´Ð¸Ð¼Ð¾ÑÑÐ¸ ÐºÐ°Ð½Ð°Ð»Ð¾Ð². ÐÐ° Ð¾ÑÐ½Ð¾Ð²Ð°Ð½Ð¸Ð¸ Ð¿Ð¾Ð»ÑÑÐµÐ½Ð½ÑÑ ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐ¾Ð² Ð¼Ð¾Ð¶Ð½Ð¾ 
Ð¿ÑÐµÐ´Ð¿Ð¾Ð»Ð¾Ð¶Ð¸ÑÑ, ÑÑÐ¾ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾-Ð°ÐºÑÐ¸Ð²Ð½ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ ENaC Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Ð562 ÑÐ¾Ð´ÐµÑÐ¶Ð°Ñ Î´ ÑÑÐ±ÑÐµÐ´Ð¸Ð½Ð¸ÑÑ.
ÐÐ°Ð»ÑÐ½ÐµÐ¹ÑÐµÐµ Ð¸Ð·ÑÑÐµÐ½Ð¸Ðµ ÑÐ¾ÑÑÐ°Ð²Ð° Ð¸ Ð¼ÐµÑÐ°Ð½Ð¸Ð·Ð¼Ð¾Ð² ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ ENaC Ð² ÑÑÐ°Ð½ÑÑÐ¾ÑÐ¼Ð¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ ÐºÐ»ÐµÑÐºÐ°Ñ ÐºÑÐ¾Ð²Ð¸ Ð°ÐºÑÑÐ°Ð»ÑÐ½Ð¾ Ð´Ð»Ñ ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐºÐ¸ Ð½Ð¾Ð²ÑÑ ÑÐ°ÑÐ³ÐµÑÐ½ÑÑ ÑÐµÑÐ°Ð¿ÐµÐ²ÑÐ¸ÑÐµÑÐºÐ¸Ñ ÑÑÑÐ°ÑÐµÐ³Ð¸Ð¹ Ð²Ð¾Ð·Ð´ÐµÐ¹ÑÑÐ²Ð¸Ñ Ð½Ð° ÑÐ°ÐºÐ¾Ð²ÑÐµ ÐºÐ»ÐµÑÐºÐ¸.
description: The given work is devoted to voltage-independent sodium channels in human leukemia cells. According to their characteristics and regulatory mechanisms they are close to the ENaC family, but not sensitive to inhibitor amiloride. The question remains about their possible subunit composition. The aim of the work was to study the possible participation of the Î´-ENaC subunit in the composition of functionally active sodium channels in K562 cells.
Using whole-cell patch-clamp method and Cytochalasine D for cytoskeleton disassembly, we recorded the activity of single actin-gated channels (in 100% of experiments), their characteristics were studied in extracellular solution containing Na+ or Li+. The single-channel conductance of some channels for Li+ was reduced in comparison to Na+ (145 mM). This characteristic is the main biophysical feature of ENaC that contains Î´-subunit (instead of Î±). Using immunofluorescence staining of K562 cells with specific antibodies, the presence of the Î´-subunit of ENaC was confirmed (along with Î±-, Î²-, Î³-ENaC).
In whole-cell experiments it was found that the extracellular addition of capsazepine, an activator of Î´ ENaC, caused the activation of sodium channels in 67% experiments. The electrophysiological characteristics of the capsazepine- 
activated channels coincided with the characteristics of actin-gated channels. The replacement of solution in the chamber with Li+-containing one resulted in decrease of current amplitude and channel conductance. Based on this fact, it can be assumed that ENaC channels in K562 cells contain Î´ subunit.
Further study of the composition and mechanisms of regulation of ENaC in transformed blood cells is relevant for the development of new targeted therapeutic strategies for cancer treatment.