These authors contributed equally.
Pharmacological profiling of the TRPV3 channel in recombinant and native assays
Article first published online: 28 APR 2014
© 2013 The British Pharmacological Society
British Journal of Pharmacology
Special Issue: Themed Section: The pharmacology of TRP channels. Guest Editors: Peter Holzer & Angelo A. Izzo
Volume 171, Issue 10, pages 2631–2644, May 2014
How to Cite
Grubisha, O., Mogg, A. J., Sorge, J. L., Ball, L.-J., Sanger, H., Ruble, C. L. A., Folly, E. A., Ursu, D. and Broad, L. M. (2014), Pharmacological profiling of the TRPV3 channel in recombinant and native assays. British Journal of Pharmacology, 171: 2631–2644. doi: 10.1111/bph.12303
- Issue published online: 28 APR 2014
- Article first published online: 28 APR 2014
- Accepted manuscript online: 15 JUL 2013 05:36AM EST
- Manuscript Accepted: 10 JUL 2013
- Manuscript Revised: 4 JUL 2013
- Manuscript Received: 20 MAR 2013
- Eli Lilly
- TRP channel;
- mouse 308 keratinocytes
Background and Purpose
Transient receptor potential vanilloid subtype 3 (TRPV3) is implicated in nociception and certain skin conditions. As such, it is an attractive target for pharmaceutical research. Understanding of endogenous TRPV3 function and pharmacology remains elusive as selective compounds and native preparations utilizing higher throughput methodologies are lacking. In this study, we developed medium-throughput recombinant and native cellular assays to assess the detailed pharmacological profile of human, rat and mouse TRPV3 channels.
Medium-throughput cellular assays were developed using a Ca2+-sensitive dye and a fluorescent imaging plate reader. Human and rat TRPV3 pharmacology was examined in recombinant cell lines, while the mouse 308 keratinocyte cell line was used to assess endogenous TRPV3 activity.
A recombinant rat TRPV3 cellular assay was successfully developed after solving a discrepancy in the published rat TRPV3 protein sequence. A medium-throughput, native, mouse TRPV3 keratinocyte assay was also developed and confirmed using genetic approaches. Whereas the recombinant human and rat TRPV3 assays exhibited similar agonist and antagonist profiles, the native mouse assay showed important differences, namely, TRPV3 activity was detected only in the presence of potentiator or during agonist synergy. Furthermore, the native assay was more sensitive to block by some antagonists.
Conclusions and Implications
Our findings demonstrate similarities but also notable differences in TRPV3 pharmacology between recombinant and native systems. These findings offer insights into TRPV3 function and these assays should aid further research towards developing TRPV3 therapies.
This article is part of a themed section on the pharmacology of TRP channels. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue-10