TY - JOUR
T1 - GPR55 ligands promote receptor coupling to multiple signalling pathways
AU - Henstridge, Christopher M.
AU - Balenga, Nariman A. B.
AU - Schroeder, Ralf
AU - Kargl, Julia K.
AU - Platzer, Wolfgang
AU - Martini, Lene
AU - Arthur, Simon
AU - Penman, June
AU - Whistler, Jennifer L.
AU - Kostenis, Evi
AU - Waldhoer, Maria
AU - Irving, Andrew J.
PY - 2010/6
Y1 - 2010/6
N2 - Background and purpose:Although GPR55 is potently activated by the endogenous lysophospholipid, L-alpha-lysophosphatidylinositol (LPI), it is also thought to be sensitive to a number of cannabinoid ligands, including the prototypic CB1 receptor antagonists AM251 and SR141716A (Rimonabant (R)). In this study we have used a range of functional assays to compare the pharmacological activity of selected cannabinoid ligands, AM251, AM281 and SR141716A with LPI in a HEK293 cell line engineered to stably express recombinant, human GPR55.Experimental approach:We evaluated Ca2+ signalling, stimulation of extracellular signal regulated kinase (ERK1/2) mitogen activated kinase MAP-kinases, induction of transcriptional regulators that are downstream of GPR55, including nuclear factor of activated T cells (NFAT), nuclear factor-kappa B (NF-kappa B) and cAMP response element binding protein (CREB), as well as receptor endocytosis. In addition, we assessed the suitability of a novel, label-free assay for GPR55 ligands that involves optical measurement of dynamic mass redistribution following receptor activation.Key results:GPR55 linked to a range of downstream signalling events and that the activity of GPR55 ligands was influenced by the functional assay employed, with differences in potency and efficacy observed.Conclusions and implications:Our data help to resolve some of the issues surrounding the pharmacology of cannabinoid ligands at GPR55 and highlight some differences in effector coupling associated with distinct GPR55 ligands.
AB - Background and purpose:Although GPR55 is potently activated by the endogenous lysophospholipid, L-alpha-lysophosphatidylinositol (LPI), it is also thought to be sensitive to a number of cannabinoid ligands, including the prototypic CB1 receptor antagonists AM251 and SR141716A (Rimonabant (R)). In this study we have used a range of functional assays to compare the pharmacological activity of selected cannabinoid ligands, AM251, AM281 and SR141716A with LPI in a HEK293 cell line engineered to stably express recombinant, human GPR55.Experimental approach:We evaluated Ca2+ signalling, stimulation of extracellular signal regulated kinase (ERK1/2) mitogen activated kinase MAP-kinases, induction of transcriptional regulators that are downstream of GPR55, including nuclear factor of activated T cells (NFAT), nuclear factor-kappa B (NF-kappa B) and cAMP response element binding protein (CREB), as well as receptor endocytosis. In addition, we assessed the suitability of a novel, label-free assay for GPR55 ligands that involves optical measurement of dynamic mass redistribution following receptor activation.Key results:GPR55 linked to a range of downstream signalling events and that the activity of GPR55 ligands was influenced by the functional assay employed, with differences in potency and efficacy observed.Conclusions and implications:Our data help to resolve some of the issues surrounding the pharmacology of cannabinoid ligands at GPR55 and highlight some differences in effector coupling associated with distinct GPR55 ligands.
KW - GPR55
KW - GPCR
KW - cannabinoid
KW - LPI
KW - CANNABINOID RECEPTOR
KW - IDENTIFICATION
KW - ACTIVATION
KW - BIOSENSOR
KW - CELLS
U2 - 10.1111/j.1476-5381.2009.00625.x
DO - 10.1111/j.1476-5381.2009.00625.x
M3 - Article
C2 - 20136841
SN - 0007-1188
VL - 160
SP - 604
EP - 614
JO - British Journal of Pharmacology
JF - British Journal of Pharmacology
IS - 3
ER -