Wang Hua, Li Tao, Lei Ming, Li Miao-ling, Ding Yin-yuan, Yang Yan, Zeng Xiao-rong
Key Laboratory of Medical Electrophysiology, Ministry of Education of China, Luzhou Medical College, Luzhou, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2012 May;28(3):214-8.
To 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.
Single 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.
Mixed 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.
The 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.
建立两性霉素B和β-七叶皂苷的穿孔膜片钳技术，并研究小电导钙激活钾通道SK2电流的钙离子调控机制。
从右心耳酶解分离单个成人心房肌细胞。穿孔电极液中使用两性霉素B和/或β-七叶皂苷。采用穿孔膜片钳技术研究人心房肌细胞中钙离子对SK2电流的调控。通过细胞内钙检测系统测量细胞内钙变化。
与单独使用150μg/ml两性霉素B或6.88μg/mlβ-七叶皂苷相比，混合穿孔电极液更容易封接细胞并激活SK2电流。此外，穿孔形成后，F340/380比值与细胞内游离钙离子浓度升高的变化一致。F340/380比值通过细胞内钙检测系统测量。
适当浓度的两性霉素B与β-七叶皂苷混合可形成稳定的全细胞膜片记录技术，适用于研究细胞内钙对SK2电流的调控。
