2015 - The backstage pass to study your favorite TRP channel
Port-a-Patch and Patchliner and SyncroPatch 384PE and SyncroPatch 384PE poster, TRP Meeting 2015 (2.2 MB)
TRP channels exhibit a large variety of functional properties and play diverse cellular and physiological roles. Due to their presence in the plasma membrane of cells from various tissues and even organelles, new biophysical properties and mechanisms of regulation are constantly discovered by academic laboratories over the years. Also, because of their relevance linked to disease and pain, TRP channels are important targets in the pharmaceutical industry. Extensive functional studies of ion channels are often compromised by technical limitations of the electrophysiological assay. Nanion’s Port-A-Patch, Patchliner and SyncroPatch 384PE are specifically designed to marry the required experimental flexibility with high-throughput and Giga-seal data quality.
In order to demonstrate reliable pharmacology combined with high-throughput electrophysiology we show data of TRPA1 activated by the agonist Supercinnamaldehyde and blocked by A-967079 in a concentration-dependent manner across a SyncroPatch 384 chip in a single experiment. With the aim to highlight experimental flexibility we present data using the internal perfusion of the SyncroPatch 384PE applying NMM (biotinylated NMethyl Maleimide) and AITC (allyl-iso-thiocyanat) to the intracellular side of the channel. Intracellular organelles have been accessed using the Port-a-Patch in a report from Chen et al. (Nature Comm., 2014). Here, they measured whole lysosomes and demonstrated how synthetic small-molecule activators can restore mutant TRPML1 channel function to rescue disease associated abnormalities in mucolipidosis type IV. Furthermore, single channel traces of TRPA1 reconstituted into liposomes were recorded at various cold temperatures and the open probability was analysed. Finally, we show data featuring the temperature control of the Patchliner (available for the SyncroPatch 384PE shortly) by exposing TRPV1 and -V3 to a range of well defined temperatures and generating the corresponding activation curves.