title: [Application of recording SK2 current in human atrial myocytes by perforated patch clamp techniques with the mix of beta-escin and amphotericin B].
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Key Laboratory of Medical Electrophysiology, Ministry of Education of China, Luzhou Medical College, Luzhou, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.forename: Ming
authors.initials: M
authors.lastname: Lei
authors.forename: Miao-ling
authors.initials: ML
authors.lastname: Li
authors.forename: Yin-yuan
authors.initials: YY
authors.lastname: Ding
authors.forename: Yan
authors.initials: Y
authors.lastname: Yang
authors.forename: Xiao-rong
authors.initials: XR
authors.lastname: Zeng
mesh_terms: D000666:Amphotericin B / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D004553:Electric Conductivity; D004928:Escin / Q000494:pharmacology*; D006801:Humans; D032383:Myocytes, Cardiac / Q000502:physiology*; D018408:Patch-Clamp Techniques; D024681:Potassium Channels, Calcium-Activated / Q000187:drug effects*
chemical_list: D024681:Potassium Channels, Calcium-Activated; D004928:Escin; D000666:Amphotericin B; D002118:Calcium