Application of recording SK2 current in human atrial myocytes by perforated patch clamp techniques with the mix of beta-escin and amphotericin B.
- Author:
Hua WANG
1
;
Tao LI
;
Ming LEI
;
Miao-ling LI
;
Yin-yuan DING
;
Yan YANG
;
Xiao-rong ZENG
Author Information
- Publication Type:Journal Article
- MeSH: Amphotericin B; pharmacology; Calcium; metabolism; Electric Conductivity; Escin; pharmacology; Humans; Myocytes, Cardiac; physiology; Patch-Clamp Techniques; Potassium Channels, Calcium-Activated; drug effects
- From: Chinese Journal of Applied Physiology 2012;28(3):214-218
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a perforated patch-clamp technology with amphotericin B and beta-escin and to research the regulation of small conductance calcium-activated potassium channel SK2 currents by calcium ions.
METHODSSingle human atrial myocytes were enzymatically isolated from the right atrial appendage. Amphotericin B and / or beta-escin were used by perforated electrode liquid. The regulation of SK2 current by calcium ions in human atrial myocytes was performed with the perforated patch-clamp technique. The intracellular calcium changes were measured by the intracellular calcium test system.
RESULTSMixed perforated electrode liquid compared with 150 microg/ml amphotericin B or 6.88 microg/ml beta-escin alone, it was easy to seal cells and activate SK2 current by the former method. Moreover, the ration of F340/380 was consistent with the change of intracellular free calcium ion concentration increase after the formation of perforation. The ration of F340/380 was measured by intracellular calcium test system.
CONCLUSIONThe appropriate concentration of amphotericin B mixed with beta-escin can form a stable whole-cell patch recording technology that is appropriate for the research of SK2 current regulation by intracellular calcium.
