14.10.2020 | Webinar: Measurement of Transient receptor potential cation (TRP) channels using the Patchliner and Port-a-Patch
Port-a-Patch and Patchliner Webinar
Date: October 14. 2020
Dr. András Horváth (Nanion Technologies; Germany)
This is an on-demand webinar from Nan]i[on and Friends 2020.
CHO cells expressing transient receptor potential cation channel V (TRPV) members 1, 3 and 4 and subfamily M member 8 were studied using our automated patch clamp systems, the Patchliner Octo (PL) and Port-a-Patch Perfusion (PaPP). During the recordings, heat, cold and/or ligand activation was performed. A classical ramp pulse-protocol (–100 mV to 100 mV) was applied.
Heat activation of TRPV1, 3, 4 channels was performed repeatedly by the heated pipetted (37-45 °C) of the PL. Interestingly ruthenium red (RR, 50 and 200 µM) was not able to prevent heat activation. Experiments involving TRPV4 were also performed on the PaPP. The cannel could be activated by heat and only partially blocked by RR. Ligand activation could be also performed on the PL (10 µM Capsaicin – TRPV1, 200 µM 2-APB – TRPV3, 100 nM GSK1016790 – TRPV4) and TRPV4 on the PaPP. In all cases the effect could be inhibited using blockers. TRPM8 channel could be repetitively activated using solution at 10°C on the PaPP at 10 °C. Capsazepine (10 µM) was used to block the activated current.
Both the PL and PaPP are powerful tools to study TRP channel physiology (both using heat activation and ligand activation) and could be used to find compounds which block the temperature and ligand response separately.