The development of automated patch clamp assays for canonical transient receptor potential channels TRPC3, 6, and 7.

摘要: The canonical transient receptor potential channel subfamily (TRPC3, TRPC6, and TRPC7) contains Ca2+ permeable non-selective cation channels that are widely expressed in a variety of tissues. There is increasing evidence implicating TRPC channels, particularly TRPC3 6, physiological pathophysiological processes, eliciting interest these as novel drug targets. Electrophysiology remains benchmark technique for measuring ion function accurately determining the pharmacological effects compounds. In this report we describe development inhibitor assays on 2 automated planar patch clamp platforms - IonWorks® Quattro™ QPatch® systems. To enable activation by carbachol, Chinese Hamster Ovary-K1 cells stably expressing muscarinic M3 were transduced with human TRPC3, or TRPC7 using BacMam viruses. 7 currents could be recorded both platforms. However, design each platform limits which assay parameters can recorded. Due to its continuous recording capabilities, QPatch capture decay response. nature inability reactivate large variation peak ability develop compound screening. IonWorks Quattro, due discontinuous sampling, did not fully currents. Quattro record from 64 per well, well was sufficiently reduced allowing medium-throughput screening assays. © Copyright 2014, Mary Ann Liebert, Inc. 2014.