The thermo- and pain-sensitive Transient Receptor Potential Melastatin 3 and 8 (TRPM3 and TRPM8) ion channels are functionally associated in the lipid rafts of the plasma membrane. We have already described that cholesterol and sphingomyelin depletion, or inhibition of sphingolipid biosynthesis decreased the TRPM8 but not the TRPM3 channel opening on cultured sensory neurons. We aimed to test the effects of lipid raft disruptors on channel activation on TRPM3- and TRPM8-expressing HEK293T cells in vitro, as well as their potential analgesic actions in TRPM3 and TRPM8 channel activation involving acute pain models in mice. CHO cell viability was examined after lipid raft disruptor treatments and their effects on channel activation on channel expressing HEK293T cells by measurement of cytoplasmic Ca2+ concentration were monitored. The effects of treatments were investigated in Pregnenolone-Sulphate-CIM-0216-evoked and icilin-induced acute nocifensive pain models in mice. Cholesterol depletion decreased CHO cell viability. Sphingomyelinase and methyl-beta-cyclodextrin reduced the duration of icilin-evoked nocifensive behavior, while lipid raft disruptors did not inhibit the activity of recombinant TRPM3 and TRPM8. We conclude that depletion of sphingomyelin or cholesterol from rafts can modulate the function of native TRPM8 receptors. Furthermore, sphingolipid cleavage provided superiority over cholesterol depletion, and this method can open novel possibilities in the management of different pain conditions.

Department of Obstetrics and Gynaecology University of Pécs Édesanyák Str 17 H 7624 Pécs Hungary

Department of Pharmacology and Pharmacotherapy Medical School University of Pécs Szigeti Str 12 H 7624 Pécs Hungary

Department of Pharmacology Faculty of Pharmacy University of Pécs Rókus Str 2 H 7624 Pécs Hungary

Department of Physiology Faculty of Medicine University of Debrecen Nagyerdei Cct 98 H 4032 Debrecen Hungary

Hungarian Research Network Chronic Pain Research Group University of Pécs Szigeti Str 12 H 7624 Pécs Hungary

Institute of Chemistry Department of Organic Chemistry University of Pannonia Egyetem Str 10 H 8200 Veszprém Hungary

Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo Namesti 2 166 10 Prague Czech Republic

National Laboratory for Drug Research and Development Magyar Tudósok Cct 2 H 1117 Budapest Hungary

PharmInVivo Ltd Szondy György Str 10 H 7629 Pécs Hungary

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