Fluorescent ligand binding reveals heterogeneous distribution of adrenoceptors and 'cannabinoid-like' receptors in small arteries.
OBJECTIVE
Pharmacological analysis of synergism or functional antagonism between different receptors commonly assumes that interacting receptors are located in the same cells. We have now investigated the distribution of alpha-adrenoceptors, beta-adrenoceptors and cannabinoid-like (GPR55) receptors in the mouse arteries.


METHODS
Fluorescence intensity from vascular tissue incubated with fluorescent ligands (alpha(1)-adrenoceptor ligand, BODIPY-FL-prazosin, QAPB; beta-adrenoceptor ligand, TMR-CGP12177; fluorescent angiotensin II; a novel diarylpyrazole cannabinoid ligand (Tocrifluor 1117, T1117) was measured with confocal microscopy. Small mesenteric and tail arteries of wild-type and alpha(1B/D)-adrenoceptor-KO mice were used.


RESULTS
T1117, a fluorescent form of the cannabinoid CB(1) receptor antagonist AM251, was a ligand for GPR55, with low affinity for CB(1) receptors. In mesenteric arterial smooth muscle cells, alpha(1A)-adrenoceptors were predominantly located in different cells from those with beta-adrenoceptors, angiotensin receptors or cannabinoid-like (GPR55) receptors. Cells with beta-adrenoceptors predominated at arterial branches. Endothelial cells expressed beta-adrenoceptors, alpha-adrenoceptors and cannabinoid-like receptors. Only endothelial alpha-adrenoceptors appeared in clusters. Adventitia was a rich source of G protein-coupled receptors (GPCRs), particularly fibroblasts and nerve tracts, where Schwann cells bound alpha-adrenoceptor, beta-adrenoceptor and CB-receptor ligands, with a mix of separate receptor locations and co-localization.


CONCLUSIONS
Within each cell type, each GPCR had a distinctive heterogeneous distribution with limited co-localization, providing a guide to the possibilities for functional synergism, and suggesting a new paradigm for synergism in which interactions may be either between cells or involve converging intracellular signalling processes.
Integrative and Systems Biology, Faculty of Biomedical and Life Sciences, West Medical Building, University of Glasgow, Glasgow, UK. c.daly@bio.gla.ac.uk
C J
CJ
Daly
R A
RA
Ross
J
J
Whyte
C M
CM
Henstridge
A J
AJ
Irving
J C
JC
McGrath
D000804:Angiotensin II / Q000378:metabolism; D000818:Animals; D001896:Boron Compounds / Q000378:metabolism; D003238:Connective Tissue / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D008024:Ligands; D008297:Male; D008638:Mesenteric Arteries / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D057054:Molecular Imaging; D015336:Molecular Probe Techniques; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D011224:Prazosin / Q000031:analogs & derivatives / Q000378:metabolism; D011412:Propanolamines / Q000378:metabolism; D011720:Pyrazoles / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011941:Receptors, Adrenergic / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D043882:Receptors, Cannabinoid / Q000378:metabolism*; D013623:Tail / Q000098:blood supply*
C547717:Adra1b protein, mouse; C547720:Adra1d protein, mouse; C412306:BODIPY-FL prazosin; D001896:Boron Compounds; D005456:Fluorescent Dyes; C526692:GPR55 protein, mouse; D008024:Ligands; D011412:Propanolamines; D011720:Pyrazoles; D011941:Receptors, Adrenergic; D018340:Receptors, Adrenergic, alpha-1; D011943:Receptors, Adrenergic, beta; D043882:Receptors, Cannabinoid; D000804:Angiotensin II; C033394:CGP 12177; D011224:Prazosin