We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
A Cell-Sparing Electric Field Stimulation Technique forHigh-Throughput Screening of Voltage-Gated Ion Channels.
- Authors
Randal M. Bugianesi; Paul R. Augustine; Karim Azer; Claude Dufresne; James Herrington; Gary S. Kath; Owen B. McManus; Christopher S. Napolitano; Alison Rush; Jeff Sachs; Neal Simpson; Michael K. Wismer; Gregory J. Kaczorowski; Robert S. Slaughter
- Abstract
The Trans Cell Layer Electrical Field Stimulation (TCL-EFS) system has been developedfor high-throughput screening (HTS) of voltage-gated ion channels in microplate format on aVoltage-Ion Probe Reader (VIPR®) platform. In this design, a wire electrode is placed above the celllayer of each filter well, and a whole plate perimeter electrode resides beneath the filter layer. Thisconfiguration allows the electrodes to be placed away from the cell layer to minimize the nearelectrode field effects on cell function and dye bleaching observed with other existing designs.Mathematical simulation indicates that the electric field at the cell layer becomes uniform as the topelectrode is raised to a position near the surface of the solution in the well. Using the TCL-EFSsystem and membrane potential fluorescence resonance energy transfer (FRET) dyes, the sensitivityof voltage-gated sodium channels to tetrodotoxin and other channel inhibitors was found to besimilar to those determined by established electrophysiological and more conventional VIPRtechniques. A good correlation was also observed with the TCL-EFS system for inhibition of Cav2.2by ω-conotoxin-GVIA and for block of Cav1.2 by known small molecule inhibitors. Thus, the TCLEFS system is suitable for both quantitative analysis and HTS of voltage-gated sodium and calciumchannels, without the liabilities of previously reported EFS methodologies.
- Publication
Assay & Drug Development Technologies, 2006, Vol 4, Issue 1, p21
- ISSN
1540-658X
- Publication type
Article