Fluorescent ligand binding reveals heterogeneous distribution of adrenoceptors and 'cannabinoid-like' receptors in small arteries

Craig J Daly, Ruth Alexandra Ross, Julie Anne Whyte, C. M. Henstridge, A J Irving, J C McGrath

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

Background and purpose: 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.

Experimental approach: 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.

Key results: T1117, a fluorescent form of the cannabinoid CB1 receptor antagonist AM251, was a ligand for GPR55, with low affinity for CB1 receptors. In mesenteric arterial smooth muscle cells, alpha(1A)-adrenoceptors were predominantly located in different cells from those with b-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 and implications: 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. British Journal of Pharmacology (2010) 159, 787-796; doi:10.1111/j.1476-5381.2009.00608.x; published online 5 February 2010

Original languageEnglish
Pages (from-to)787-796
Number of pages10
JournalBritish Journal of Pharmacology
Volume159
Issue number4
Early online date5 Feb 2010
DOIs
Publication statusPublished - Feb 2010

Keywords

  • vascular smooth muscle
  • adrenoceptor
  • alpha(1B/D)-adrenoceptor knockout
  • fluorescent ligand binding
  • confocal microscopy
  • angiotensin
  • cannabinoid
  • adventitia
  • endothelium
  • smooth-muscle-cells
  • alpha(1D)-adrenergic receptors
  • cardiovascular-disease
  • mesenteric-arteries
  • surface expression
  • angiotensin-II
  • pharmacology
  • anandamide
  • subtypes
  • GPR55
  • a1B/D-adrenoceptor knockout

Cite this

Fluorescent ligand binding reveals heterogeneous distribution of adrenoceptors and 'cannabinoid-like' receptors in small arteries. / Daly, Craig J; Ross, Ruth Alexandra; Whyte, Julie Anne; Henstridge, C. M.; Irving, A J; McGrath, J C.

In: British Journal of Pharmacology, Vol. 159, No. 4, 02.2010, p. 787-796.

Research output: Contribution to journalArticle

Daly, Craig J ; Ross, Ruth Alexandra ; Whyte, Julie Anne ; Henstridge, C. M. ; Irving, A J ; McGrath, J C. / Fluorescent ligand binding reveals heterogeneous distribution of adrenoceptors and 'cannabinoid-like' receptors in small arteries. In: British Journal of Pharmacology. 2010 ; Vol. 159, No. 4. pp. 787-796.
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N2 - Background and purpose: 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.Experimental approach: 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.Key results: T1117, a fluorescent form of the cannabinoid CB1 receptor antagonist AM251, was a ligand for GPR55, with low affinity for CB1 receptors. In mesenteric arterial smooth muscle cells, alpha(1A)-adrenoceptors were predominantly located in different cells from those with b-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 and implications: 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. British Journal of Pharmacology (2010) 159, 787-796; doi:10.1111/j.1476-5381.2009.00608.x; published online 5 February 2010

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KW - endothelium

KW - smooth-muscle-cells

KW - alpha(1D)-adrenergic receptors

KW - cardiovascular-disease

KW - mesenteric-arteries

KW - surface expression

KW - angiotensin-II

KW - pharmacology

KW - anandamide

KW - subtypes

KW - GPR55

KW - a1B/D-adrenoceptor knockout

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