1P367 Whole-cell patch-clamp analysis of large conductance, stretch-activated, Ca^-activated K^+ channel in chick cultured ventricular myocytes(14. Ion channels and receptors,Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

2006 ◽  
Vol 46 (supplement2) ◽  
pp. S238
Author(s):  
Masahiro Kishio ◽  
Keiji Naruse
Keyword(s):  
Ion Channels ◽  
Patch Clamp ◽  
Poster Session ◽  
Ventricular Myocytes ◽  
K Channel ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Meeting Program ◽  
Whole Cell Patch Clamp

scholarly journals A Novel Membrane Potential-Sensitive Fluorescent Dye Improves Cell-Based Assays for Ion Channels

2002 ◽  
Vol 7 (1) ◽  
pp. 79-85 ◽  
Author(s):  
Deborah F. Baxter ◽  
Martin Kirk ◽  
Amy F. Garcia ◽  
Alejandra Raimondi ◽  
Mats H. Holmqvist ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Ion Channels ◽  
Patch Clamp ◽  
High Throughput Screening ◽  
Fluorescent Dye ◽  
Patch Clamp Recording ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp ◽  
Electrophysiological Studies

The study of ion channel-mediated changes in membrane potential using the conventional bisoxonol fluorescent dye DiBAC4(3) has several limitations, including a slow onset of response and multistep preparation, that limit both the fidelity of the results and the throughput of membrane potential assays. Here, we report the characterization of the FLIPR Membrane Potential Assay Kit (FMP) in cells expressing voltage- and ligand-gated ion channels. The steady-state and kinetics fluorescence properties of FMP were compared with those of DiBAC4(3), using both FLIPR and whole-cell patch-clamp recording. Our experiments with the voltage-gated K+ channel, hElk-1, revealed that FMP was 14-fold faster than DiBAC4(3) in response to depolarization. On addition of 60 mM KCl, the kinetics of fluorescence changes of FMP using FLIPR were identical to those observed in the electrophysiological studies using whole-cell current clamp. In addition, KCl concentration-dependent increases in FMP fluorescence correlated with the changes of membrane potential recorded in whole-cell patch clamp. In studies examining vanilloid receptor-1, a ligand-gated nonselective cation channel, FMP was superior to DiBAC4(3) with respect to both kinetics and amplitude of capsaicin-induced fluorescence changes. FMP has also been used to measure the activation of KATP1 and hERG.2 Thus this novel membrane potential dye represents a powerful tool for developing high-throughput screening assays for ion channels.

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