Abstract
Aim:
Pirt is a two-transmembrane domain protein that regulates the function of a variety of ion channels. Our previous study indicated that Pirt acts as a positive endogenous regulator of the TRPM8 channel. The aim of this study was to investigate the mechanism underlying the regulation of TRPM8 channel by Pirt.
Methods:
HEK293 cells were transfected with TRPM8+Pirt or TRPM8 alone. Menthol (1 mmol/L) was applied through perfusion to induce TRPM8-mediated voltage-dependent currents, which were recorded using a whole-cell recording technique. PIP2 (10 μmol/L) was added into the electrode pipettes (PI was taken as a control). Additionally, cell-attached single-channel recordings were conducted in CHO cells transfected with TRPM8+Pirt or TRPM8 alone, and menthol (1 mmol/L) was added into the pipette solution.
Results:
Either co-transfection with Pirt or intracellular application of PIP2 (but not PI) significantly enhanced menthol-induced TRPM8 currents. Furthermore, Pirt and PIP2 synergistically modulated menthol-induced TRPM8 currents. Single-channel recordings revealed that co-transfection with Pirt significantly increased the single channel conductance.
Conclusion:
Pirt and PIP2 synergistically enhance TRPM8 channel activity, and Pirt regulates TRPM8 channel activity by increasing the single channel conductance.
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Acknowledgements
This work was supported by the National Science Foundation of China to Zong-xiang TANG (31271181 and 31471007), by Oversea, Hong Kong & Macao Scholars Collaborated Researching Fund to Xin-zhong DONG and Zong-xiang TANG (31328012), from the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), sponsored by “Qing Lan Project” in Jiangsu Province and from the Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor, Nanjing University of Chinese Medicine, China. This work was also supported by Cooperative Innovation Center for Molecular Target New Drug Study, Research and Innovation Project of Hunan Province (CX2015B553) to Min TANG
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Tang, M., Wu, Gy., Dong, Xz. et al. Phosphoinositide interacting regulator of TRP (Pirt) enhances TRPM8 channel activity in vitro via increasing channel conductance. Acta Pharmacol Sin 37, 98–104 (2016). https://doi.org/10.1038/aps.2015.110
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DOI: https://doi.org/10.1038/aps.2015.110
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