Journal
EUROPEAN JOURNAL OF PHARMACOLOGY
Volume 805, Issue -, Pages 1-13Publisher
ELSEVIER
DOI: 10.1016/j.ejphar.2017.03.046
Keywords
G protein-coupled receptor (GPCR); G protein; beta-arrestin; Neurotensin receptor 1; Neurotensin; Neuromedin N
Categories
Funding
- Canadian Institute of Health Research (CIHR) [FDN-148413]
- National Science and Engineering Research Council of Canada (NSERC) [CRDPJ-399680]
- FRQ-S
- Institut de Pharmacologie de Sherbrooke (IPS)
- Centre d'Excellence en Neurosciences de l'Universite de Sherbrooke (CNS)
- Canada Research Chair in Neurophysiopharmacology of Chronic Pain
Ask authors/readers for more resources
The human neurotensin 1 receptor (hNTS1) is a G protein-coupled receptor involved in many physiological functions, including analgesia, hypothermia, and hypotension. To gain a better understanding of which signaling pathways or combination of pathways are linked to NTS1 activation and function, we investigated the ability of activated hNTS1, which was stably expressed by CHO-K1 cells, to directly engage G proteins, activate second messenger cascades and recruit (beta-arrestins. Using BRET-based biosensors, we found that neurotensin (NT), NT(8-13) and neuromedin N (NN) activated the G alpha(q)-, G alpha(OA)-, and G alpha(13)-protein signaling pathways as well as the recruitment of beta-arrestins 1 and 2. Using pharmacological inhibitors, we further demonstrated that all three ligands stimulated the production of inositol phosphate and modulation of cAMP accumulation along with ERK1/2 activation. Interestingly, despite the functional coupling to G alpha(i1) and Ga-OA, NT was found to produce higher levels of cAMP in the presence of pertussis toxin, supporting that hNTS1 activation leads to cAMP accumulation in a Ga-s-dependent manner. Additionally, we demonstrated that the full activation of ERK1/2 required signaling through both a PTX-sensitive G(i/o)-c-Src signaling pathway and PLC beta-DAG-PKC-Raf-1-dependent pathway downstream of G(q). Finally, the whole-cell integrated signatures monitored by the cell-based surface plasmon resonance and changes in the electrical impedance of a confluent cell monolayer led to identical phenotypic responses between the three ligands. The characterization of the hNTS1-mediated cellular signaling network will be helpful to accelerate the validation of potential NTS1 biased ligands with an improved therapeutic/adverse effect profile.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available