Activation of transient receptor potential V1 (TRPV1) is critical for pain sensation. There are some indications that the function of TRPV1 can be modulated through its communication with the μ-opioid receptor (MOR) and Toll-like receptor 4 (TLR4). In addition, cross-talk between signaling cascades regulated by MOR and TLR4 has also been noted. Nevertheless, the mutual interactions between all these receptors and their signaling pathways are still not well understood. The main goal of the present project is to investigate the effects of different specific ligands on the behavior and function of these three membrane-bound receptors, which participate in processing of nociceptive stimuli. Using advanced fluorescence microscopy techniques such as FRAP (fluroescence recovery after photo-bleaching) and FLIM-FRET (fluorescence lifetime imaging-fluorescence resonance energy transfer) or BRET (bioluminescence resonance energy transfer) we will monitor receptor lateral mobility in the plasma membrane, dynamics of their internalization and relevant protein-protein interactions under controlled in vitro conditions. The role of key molecular partners of these receptors (e.g., G proteins, β-arrestins or GRKs) will also be explored. For this purpose, selected molecules will be knock-downed by specific siRNAs. Results of this research may help to unravel the details of the molecular interactions between MOR, TLR4 and TRPV1.
Drastichova Z., Skrabalova J., Jedelsky P., Neckar J., Kolar F. and Novotny J. (2012) Global changes in the rat heart proteome induced by prolonged morphine treatment and withdrawal. PLoS One 7:e47167.
Ujcikova H., Brejchova J., Vosahlikova M., Kagan D., Dlouha K., Sykora J., Merta L., Drastichova Z., Novotny J., Ostasov P., Roubalova L., Hof M. and Svoboda P. (2014) Opioid-receptor signaling cascades in rat cerebral cortex and model cell lines: the role of plasma membrane structure. Physiol. Res. 63(Suppl. 1):S165-176.
Pacesova D., Volfova B., Cervena K., Hejnova L., Novotny J. and Bendova Z. (2015) Acute morphine affects the rat circadian clock via rhythms of physphorylated ERK1/2 and GSK3β kinases and Per1 expression in the rat suprachiasmatic nucleus. Br. J. Pharmacol. 172:3638-3649.
Melkes B., Hejnova L. and Novotny J. (2016) Biased μ-opioid receptor agonists diversely regulate lateral mobility and functional coupling of the receptor to its cognate G proteins. Naunyn-Schmiedeberg´s Arch. Pharmacol. 389:1289-1300.
Hejnova L., Skrabalova J. and Novotny J. (2017) Prolonged morphine treatment alters expression and plasma membrane distribution of β-adrenergic receptors and some other components of their signaling system in rat cerebral cortex. J. Mol. Neurosci. 63:364-376.
Czech Science Foundation (19-03295S)
110-111520
Deadline is closed