title: ÐÐ·ÑÑÐµÐ½Ð¸Ðµ Ð±Ð¸Ð¾ÑÐ¸Ð·Ð¸ÑÐµÑÐºÐ¸Ñ Ñ Ð°ÑÐ°ÐºÑÐµÑÐ¸ÑÑÐ¸Ðº Ð¼ÐµÑ Ð°Ð½Ð¾ÑÑÐ²ÑÑÐ²Ð¸ÑÐµÐ»ÑÐ½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² PIEZO1 Ð² ÐºÐ»ÐµÑÐºÐ°Ñ K562 Ð² Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ Ð´Ð²ÑÑ Ð²Ð°Ð»ÐµÐ½ÑÐ½ÑÑ ÐºÐ°ÑÐ¸Ð¾Ð½Ð¾Ð²
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: Study of biophysical characteristics of mechanosensitive Piezo1 channels in K562 cells in the presence of divalent cations
Creators: name: ÐÐ¸ÑÐ¸Ð»Ð»Ð¾Ð²Ð°, ÐÐ¾Ð»Ð¸Ð½Ð°
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: 2023
Subjects: Ð¼ÐµÑ Ð°Ð½Ð¾ÑÑÐ²ÑÑÐ²Ð¸ÑÐµÐ»ÑÐ½ÑÐµ Ð¸Ð¾Ð½Ð½ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ
Ð¼ÐµÑÐ¾Ð´ Ð»Ð¾ÐºÐ°Ð»ÑÐ½Ð¾Ð¹ ÑÐ¸ÐºÑÐ°ÑÐ¸Ð¸ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»Ð° (patch-clamp)
ÑÐµÐ¼ÐµÐ¹ÑÑÐ²Ð¾ PIEZO
ÑÐµÐ»ÐµÐºÑÐ¸Ð²Ð½ÑÐ¹ Ñ Ð¸Ð¼Ð¸ÑÐµÑÐºÐ¸Ð¹ Ð°ÐºÑÐ¸Ð²Ð°ÑÐ¾Ñ Yoda1
Ð´Ð²ÑÑ Ð²Ð°Ð»ÐµÐ½ÑÐ½ÑÐµ ÐºÐ°ÑÐ¸Ð¾Ð½Ñ
ÐºÐ»ÐµÑÐºÐ¸ Ð¼Ð¸ÐµÐ»Ð¾Ð¸Ð´Ð½Ð¾Ð¹ Ð»ÐµÐ¹ÐºÐµÐ¼Ð¸Ð¸ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ°
mechanosensitive ion channels
patch-clamp method
PIEZO family
selective chemical activator Yoda1
divalent cations
human myeloid leukemia cells

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¸Ð·ÑÑÐµÐ½Ð¸Ñ Ð²ÐºÐ»Ð°Ð´Ð° ÐºÐ°ÑÐ¸Ð¾Ð½Ð¾Ð² Na^+, Mg^(2+) Ð¸ Ca^(2+) Ð² Ð¿ÑÐ¾Ð²Ð¾Ð´ÑÑÐ¸Ðµ ÑÐ²Ð¾Ð¹ÑÑÐ²Ð° Ð¼ÐµÑÐ°Ð½Ð¾ÑÑÐ²ÑÑÐ²Ð¸ÑÐµÐ»ÑÐ½ÑÑ ÐºÐ°ÑÐ¸Ð¾Ð½-ÑÐµÐ»ÐµÐºÑÐ¸Ð²Ð½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Piezo1 Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Ð¼Ð¸ÐµÐ»Ð¾Ð¸Ð´Ð½Ð¾Ð¹ Ð»ÐµÐ¹ÐºÐµÐ¼Ð¸Ð¸ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ° Ð562. ÐÑÑÐ»ÐµÐ´ÑÐµÐ¼ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ ÑÐ²Ð»ÑÑÑÑÑ Ð²Ð°Ð¶Ð½ÑÐ¼Ð¸ Ð¼ÐµÑÐ°Ð½Ð¾ÑÑÐ°Ð½ÑÐ´ÑÐºÑÐ¾ÑÐ°Ð¼Ð¸ Ð² Ð¿ÑÐ¾ÑÐµÑÑÐ°Ñ ÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¹ ÑÐ¸Ð³Ð½Ð°Ð»Ð¸Ð·Ð°ÑÐ¸Ð¸, Ð¼Ð¾Ð³ÑÑ Ð°ÐºÑÐ¸Ð²Ð¸ÑÐ¾Ð²Ð°ÑÑÑÑ Ð² Ð¾ÑÐ²ÐµÑ Ð½Ð° ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÐµ Ð¼ÐµÑÐ°Ð½Ð¸ÑÐµÑÐºÐ¸Ðµ Ð²Ð¾Ð·Ð´ÐµÐ¹ÑÑÐ²Ð¸Ñ, Ð° ÑÐ°ÐºÐ¶Ðµ ÑÐµÐ»ÐµÐºÑÐ¸Ð²Ð½ÑÐ¼Ð¸ ÑÐ¸Ð¼Ð¸ÑÐµÑÐºÐ¸Ð¼Ð¸ Ð°Ð³Ð¾Ð½Ð¸ÑÑÐ°Ð¼Ð¸ (Yoda1, Jedi2).Â  Ð¦ÐµÐ»ÑÑ ÑÐ°Ð±Ð¾ÑÑ ÑÐ²Ð»ÑÐ»Ð¾ÑÑ Ð¸Ð·ÑÑÐµÐ½Ð¸Ðµ Ð±Ð¸Ð¾ÑÐ¸Ð·Ð¸ÑÐµÑÐºÐ¸Ñ ÑÐ°ÑÐ°ÐºÑÐµÑÐ¸ÑÑÐ¸Ðº Yoda1-Ð°ÐºÑÐ¸Ð²Ð¸ÑÑÐµÐ¼ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Piezo1 Ð² ÐºÐ»ÐµÑÐºÐ°Ñ Ð562 Ð² Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ ÑÐ¾Ð»ÑÐºÐ¾ ÐºÐ°ÑÐ¸Ð¾Ð½Ð¾Ð² Na^+ Ð¸ Ð¿ÑÐ¸ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾Ð¼ Ð´Ð¾Ð±Ð°Ð²Ð»ÐµÐ½Ð¸Ð¸ ÐºÐ°ÑÐ¸Ð¾Ð½Ð¾Ð² Ca^(2+)Ð¸/Ð¸Ð»Ð¸ Mg^(2+). Ð ÐºÐ¾Ð½ÑÐ¸Ð³ÑÑÐ°ÑÐ¸Ð¸ whole-cell Ð¼ÐµÑÐ¾Ð´Ð° patch-clamp Ð·Ð°ÑÐµÐ³Ð¸ÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð»Ð¸ Ð¾Ð´Ð¸Ð½Ð¾ÑÐ½ÑÐµ ÐºÐ°Ð½Ð°Ð»Ñ Piezo1, Ð°ÐºÑÐ¸Ð²Ð¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÐµ Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ Yoda1 Ð² ÑÑÐ°Ð½Ð´Ð°ÑÑÐ½Ð¾Ð¼ Ð²Ð½ÐµÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¼ ÑÐ°ÑÑÐ²Ð¾ÑÐµ, ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐµÐ¼ (Ð² Ð¼Ð): 145 NaCl, 2 CaCl2, 1 MgCl2. ÐÑÐ¾Ð²Ð¾Ð´Ð¸Ð¼Ð¾ÑÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² ÑÐ¾ÑÑÐ°Ð²Ð¸Ð»Ð° Ð¾ÐºÐ¾Ð»Ð¾ 19 Ð¿Ð¡Ð¼. ÐÐ±Ð½Ð°ÑÑÐ¶Ð¸Ð»Ð¸, ÑÑÐ¾ Ð¿ÑÐ¾Ð²Ð¾Ð´Ð¸Ð¼Ð¾ÑÑÑ Yoda1-Ð¸Ð½Ð´ÑÑÐ¸ÑÑÐµÐ¼ÑÑ Piezo1 Ð² Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ ÑÐ¾Ð»ÑÐºÐ¾ Ð¸Ð¾Ð½Ð¾Ð² Na^+ (~30 Ð¿Ð¡Ð¼) Ð·Ð½Ð°ÑÐ¸ÑÐµÐ»ÑÐ½Ð¾ Ð¿ÑÐµÐ²ÑÑÐ°Ð»Ð° Ð·Ð½Ð°ÑÐµÐ½Ð¸Ñ, ÑÐ°ÑÐ°ÐºÑÐµÑÐ½ÑÐµ Ð´Ð»Ñ Piezo1 Ð² ÑÑÐ°Ð½Ð´Ð°ÑÑÐ½Ð¾Ð¼ ÑÐ°ÑÑÐ²Ð¾ÑÐµ. ÐÐ¾Ð±Ð°Ð²Ð»ÐµÐ½Ð¸Ðµ Ð¾Ð´Ð½Ð¾Ð³Ð¾ Ð¸Ð· Ð´Ð²ÑÑÐ²Ð°Ð»ÐµÐ½ÑÐ½ÑÑ ÐºÐ°ÑÐ¸Ð¾Ð½Ð¾Ð² Ca^(2+)Â  Ð¸Ð»Ð¸ Mg^(2+) Ð²Ð¾ Ð²Ð½ÐµÐºÐ»ÐµÑÐ¾ÑÐ½ÑÐ¹ ÑÐ°ÑÑÐ²Ð¾Ñ Ð¿ÑÐ¸Ð²Ð¾Ð´Ð¸Ð»Ð¾ Ðº ÑÐ½Ð¸Ð¶ÐµÐ½Ð¸Ñ Ð¿ÑÐ¾Ð²Ð¾Ð´Ð¸Ð¼Ð¾ÑÑÐ¸ Ð´Ð¾ 20 â 22 Ð¿Ð¡Ð¼. Ð Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ Ð¾Ð±Ð¾Ð¸Ñ Ð´Ð²ÑÑÐ²Ð°Ð»ÐµÐ½ÑÐ½ÑÑ ÐºÐ°ÑÐ¸Ð¾Ð½Ð¾Ð² Ð¿ÑÐ¾Ð²Ð¾Ð´Ð¸Ð¼Ð¾ÑÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² ÑÐ½Ð¸Ð¶Ð°Ð»Ð°ÑÑ Ð´Ð¾ Ð¸ÑÑÐ¾Ð´Ð½ÑÑ Ð·Ð½Ð°ÑÐµÐ½Ð¸Ð¹ (19 Ð¿Ð¡Ð¼). ÐÐ° Ð¾ÑÐ½Ð¾Ð²Ð°Ð½Ð¸Ð¸ Ð¿Ð¾Ð»ÑÑÐµÐ½Ð½ÑÑ ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐ¾Ð² Ð¼Ð¾Ð¶Ð½Ð¾ ÑÐ´ÐµÐ»Ð°ÑÑ Ð²ÑÐ²Ð¾Ð´, ÑÑÐ¾ ÐºÐ°Ð¶Ð´ÑÐ¹ Ð¸Ð· Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð½ÑÑ ÐºÐ°ÑÐ¸Ð¾Ð½Ð¾Ð² Ð²Ð½Ð¾ÑÐ¸Ñ Ð²ÐºÐ»Ð°Ð´ Ð² Ð¿ÑÐ¾Ð²Ð¾Ð´ÑÑÐ¸Ðµ ÑÐ²Ð¾Ð¹ÑÑÐ²Ð° ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Piezo1. ÐÐ¾Ð»ÑÑÐµÐ½Ð½ÑÐµ Ð´Ð°Ð½Ð½ÑÐµ Ð°ÐºÑÑÐ°Ð»ÑÐ½Ñ Ð´Ð»Ñ Ð¿Ð¾Ð½Ð¸Ð¼Ð°Ð½Ð¸Ñ Ð¿ÑÐ¾Ð²Ð¾Ð´ÑÑÐ¸Ñ ÑÐ²Ð¾Ð¹ÑÑÐ² Ð¼ÐµÑÐ°Ð½Ð¾ÑÑÐ²ÑÑÐ²Ð¸ÑÐµÐ»ÑÐ½ÑÑ ÐºÐ°Ð½Ð°Ð»Ð¾Ð² Ð¸ Ð¸Ñ ÑÐ¾Ð»Ð¸ Ð² ÑÐµÐ³ÑÐ»ÑÑÐ¸Ð¸ ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÑ ÑÐ¸Ð·Ð¸Ð¾Ð»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¸Ñ Ð¿ÑÐ¾ÑÐµÑÑÐ¾Ð², Ð² ÑÐ¾Ð¼ ÑÐ¸ÑÐ»Ðµ ÑÐ²ÑÐ·Ð°Ð½Ð½ÑÑ Ñ Ð¼ÐµÑÐ°Ð½Ð¸ÑÐµÑÐºÐ¸Ð¼Ð¸ Ð²Ð¾Ð·Ð´ÐµÐ¹ÑÑÐ²Ð¸ÑÐ¼Ð¸ Ð½Ð° ÐºÐ»ÐµÑÐºÑ.
description: The given work is devoted to the study of the contribution ofÂ  Na^+, Mg^(2+) and Ca^(2+) cations to the conductive properties of mechanosensitive Piezo1 cation-selective channels in human myeloid leukemia cells K562. The studied channels are important mechanotransducers in the processes of cell signaling, can be activated in response to various mechanical stimuli, as well as selective chemical agonists (Yoda1, Jedi2). The aim of this work was to study the biophysical characteristics of Yoda1-activated Piezo1 channels in K562 cells in the presence of only Na^+ and sequential addition of Mg^(2+) and/or Ca^(2+) cations. In the whole-cell configuration of the patch-clamp method, single Piezo1 channels activated with Yoda1 in a standard extracellular solution containing (in mM): 145 NaCl, 2 CaCl2, 1 MgCl2 were registered. The conductance of the channels was about 19 pS. It was found that the conductance of Yoda1-induced Piezo1 in the presence of only Na^+Â  ions (~30 pS) significantly exceeded the values characteristic of Piezo1 in the standard solution. The addition of one of the divalent cationsÂ  Ca^(2+)Â  or Mg^(2+)Â  cations to the extracellular solution led to a conductance decrease to 20â22 pS. In the presence of both divalent cations, the channel conductance decreased to the initial values (19 pS). Based on the results obtained, it can be concluded that each of the studied cations contributes to the conductive properties of the Piezo1 channels. The data obtained are relevant for understanding the conductive properties of mechanosensitive channels and their role in the regulation of various physiological processes, including those associated with mechanical action on the cell.