A simple method for fast temperature changes and its application to thermal activation of TRPV1 ion channels
Article
-
- Overview
-
- Research
-
- Identity
-
- Additional Document Info
-
- View All
-
Overview
abstract
-
Background: Thermally activated ion channels function as molecular thermometers and participate in other physiological important functions. The mechanism by which they acquire their exquisite temperature sensitivity is unknown and is currently an area of intense research. For this reason, there is a need for diverse methods to deliver controlled temperature stimuli. New method: We have developed a simple, inexpensive and reliable method to deliver temperature pulses to small volumes surrounding the recording area, which can be either a patch-clamp pipette containing a cell-free membrane with thermally activated channels or a whole cell attached to a pipette. Results: Here we developed a micro-heater based on resistive heating of a copper filament enclosed in a glass capillary that is capable of delivering fast and localized temperature changes. We validated the performance of the micro-heaters by analyzing the heat-induced activation of TRPV1 thermoTRP channels recorded in inside-out patches and demonstrate the use of the micro-heaters. Comparison with existing method(s): The micro-heaters we introduce here are compact, easy to fabricate and to operate. In contrast with bulk solution heaters commercially available, our method is extremely affordable and simple to operate. To the best of our knowledge there are no other similar, commercially available heating methods. Conclusions: The micro-heater method is simple and should provide a straightforward and rapid experimental tool to study mechanisms in thermally activated ion channels. © 2015 Elsevier B.V.
publication date
funding provided via
published in
Research
keywords
-
Heat activation; Ion channels; Patch-clamp; Resistive heating; ThermoTRPs; TRPV1 channel copper; glass; vanilloid receptor 1; electrolyte; solution and solubility; Trpv1 protein, rat; vanilloid receptor; Article; electrophysiology; heat sensitivity; heating; human; human cell; inside out patch clamp; nonhuman; priority journal; rat; temperature; temperature dependence; thermal activation; thermal conductivity; whole cell patch clamp; animal; computer simulation; devices; electrical equipment; equipment design; genetic transfection; genetics; HEK293 cell line; membrane potential; metabolism; patch clamp technique; physiology; procedures; solution and solubility; stimulation; theoretical model; validation study; Animals; Computer Simulation; Copper; Electrical Equipment and Supplies; Electrolytes; Equipment Design; Glass; HEK293 Cells; Humans; Membrane Potentials; Models, Theoretical; Patch-Clamp Techniques; Physical Stimulation; Rats; Solutions; Temperature; Transfection; TRPV Cation Channels
Identity
Digital Object Identifier (DOI)
PubMed ID
Additional Document Info
start page
end page
volume
issue