Perforated whole-cell patch recording of L-type calcium current with β-escin in osteoblasts
10.3969/j.issn.1000-4718.2015.06.033
- VernacularTitle:一种有效记录成骨细胞 L 型钙通道电流的穿孔全细胞膜片钳方法
- Author:
Wenwei WANG
;
Xiaoyan QI
;
Xuemei ZHANG
- Publication Type:Journal Article
- Keywords:
Osteoblasts;
L-type calcium current;
Perforated whole-cell patch-clamp;
β-escin
- From:
Chinese Journal of Pathophysiology
2015;(6):1150-1152
- CountryChina
- Language:Chinese
-
Abstract:
[ ABSTRACT] AIM:To establish a perforated whole-cell patch-clamp technique withβ-escin to record L-type cal-cium current (ICa,L) in osteoblasts.METHODS:ROS 17/2.8 is a rat osteoblast-like osteosarcoma cell line.β-escin was applied to the pipette solution to permeabilize the cell membrane and the perforated patch recording mode was obtained. RESULTS:β-escin at concentration of 50μmol/L easily permeabilized the cell membrane and obtained a perforated patch recording mode in 2~7 min.This technique prevented ICa,L rundown and preserved cytoplasmic signaling pathways.CON-CLUSION:β-escin may be used as an alternative ionophore for perforated patch-clamp studies in osteoblasts and results in minimal rundown that could facilitate recordings of ICa,L in osteoblasts.