G-protein βγ subunits in vasorelaxing and anti-endothelinergic effects of calcitonin gene-related peptide

M J P M T Meens, N J A Mattheij, P B van Loenen, L J A Spijkers, P Lemkens, J Nelissen, M G Compeer, A E Alewijnse, J G R De Mey

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

BACKGROUND AND PURPOSE: Calcitonin gene-related peptide (CGRP) has been proposed to relax vascular smooth muscle cells (VSMC) via cAMP and can promote dissociation of endothelin-1 (ET-1) from ET(A) receptors. The latter is not mimicked by other stimuli of adenylate cyclases. Therefore, we evaluated the involvement of G-protein βγ subunits (Gβγ) in the arterial effects of CGRP receptor stimulation.

EXPERIMENTAL APPROACH: To test the hypothesis that instead of α subunits of G-proteins (Gαs), Gβγ mediates the effects of CGRP receptor activation, we used (i) rat isolated mesenteric resistance arteries (MRA), (ii) pharmacological modulators of cyclic nucleotides; and (iii) low molecular weight inhibitors of the functions of Gβγ, gallein and M119. To validate these tools with respect to CGRP receptor function, we performed organ bath studies with rat isolated MRA, radioligand binding on membranes from CHO cells expressing human CGRP receptors and cAMP production assays in rat cultured VSMC.

KEY RESULTS: In isolated arteries contracted with K(+) or ET-1, IBMX (PDE inhibitor) increased sodium nitroprusside (SNP)- and isoprenaline (ISO)- but not CGRP-induced relaxations. While fluorescein (negative control) was without effects, gallein increased binding of [(125) I]-CGRP in the absence and presence of GTPγS. Gallein also increased CGRP-induced cAMP production in VSMC. Despite these stimulating effects, gallein and M119 selectively inhibited the relaxing and anti-endothelinergic effects of CGRP in isolated arteries while not altering contractile responses to K(+) or ET-1 or relaxing responses to ISO or SNP.

CONCLUSION AND IMPLICATIONS: Activated CGRP receptors induce cyclic nucleotide-independent relaxation of VSMC and terminate arterial effects of ET-1 via Gβγ.

OriginalsprogEngelsk
TidsskriftBritish Journal of Pharmacology
Vol/bind166
Udgave nummer1
Sider (fra-til)297-308
ISSN0007-1188
DOI
StatusUdgivet - 2012
Udgivet eksterntJa

Fingeraftryk

Calcitonin Gene-Related Peptide Receptors
Calcitonin Gene-Related Peptide
Protein Subunits
Endothelin-1
Vascular Smooth Muscle
Nitroprusside
Isoproterenol
1-Methyl-3-isobutylxanthine
CHO Cells
Fluorescein
Adenylyl Cyclases
Molecular Weight
Membranes
gallein

Citer dette

Meens, M J P M T ; Mattheij, N J A ; van Loenen, P B ; Spijkers, L J A ; Lemkens, P ; Nelissen, J ; Compeer, M G ; Alewijnse, A E ; De Mey, J G R. / G-protein βγ subunits in vasorelaxing and anti-endothelinergic effects of calcitonin gene-related peptide. I: British Journal of Pharmacology. 2012 ; Bind 166, Nr. 1. s. 297-308.
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abstract = "BACKGROUND AND PURPOSE: Calcitonin gene-related peptide (CGRP) has been proposed to relax vascular smooth muscle cells (VSMC) via cAMP and can promote dissociation of endothelin-1 (ET-1) from ET(A) receptors. The latter is not mimicked by other stimuli of adenylate cyclases. Therefore, we evaluated the involvement of G-protein βγ subunits (Gβγ) in the arterial effects of CGRP receptor stimulation.EXPERIMENTAL APPROACH: To test the hypothesis that instead of α subunits of G-proteins (Gαs), Gβγ mediates the effects of CGRP receptor activation, we used (i) rat isolated mesenteric resistance arteries (MRA), (ii) pharmacological modulators of cyclic nucleotides; and (iii) low molecular weight inhibitors of the functions of Gβγ, gallein and M119. To validate these tools with respect to CGRP receptor function, we performed organ bath studies with rat isolated MRA, radioligand binding on membranes from CHO cells expressing human CGRP receptors and cAMP production assays in rat cultured VSMC.KEY RESULTS: In isolated arteries contracted with K(+) or ET-1, IBMX (PDE inhibitor) increased sodium nitroprusside (SNP)- and isoprenaline (ISO)- but not CGRP-induced relaxations. While fluorescein (negative control) was without effects, gallein increased binding of [(125) I]-CGRP in the absence and presence of GTPγS. Gallein also increased CGRP-induced cAMP production in VSMC. Despite these stimulating effects, gallein and M119 selectively inhibited the relaxing and anti-endothelinergic effects of CGRP in isolated arteries while not altering contractile responses to K(+) or ET-1 or relaxing responses to ISO or SNP.CONCLUSION AND IMPLICATIONS: Activated CGRP receptors induce cyclic nucleotide-independent relaxation of VSMC and terminate arterial effects of ET-1 via Gβγ.",
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volume = "166",
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Meens, MJPMT, Mattheij, NJA, van Loenen, PB, Spijkers, LJA, Lemkens, P, Nelissen, J, Compeer, MG, Alewijnse, AE & De Mey, JGR 2012, 'G-protein βγ subunits in vasorelaxing and anti-endothelinergic effects of calcitonin gene-related peptide', British Journal of Pharmacology, bind 166, nr. 1, s. 297-308. https://doi.org/10.1111/j.1476-5381.2011.01774.x

G-protein βγ subunits in vasorelaxing and anti-endothelinergic effects of calcitonin gene-related peptide. / Meens, M J P M T; Mattheij, N J A; van Loenen, P B; Spijkers, L J A; Lemkens, P; Nelissen, J; Compeer, M G; Alewijnse, A E; De Mey, J G R.

I: British Journal of Pharmacology, Bind 166, Nr. 1, 2012, s. 297-308.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - G-protein βγ subunits in vasorelaxing and anti-endothelinergic effects of calcitonin gene-related peptide

AU - Meens, M J P M T

AU - Mattheij, N J A

AU - van Loenen, P B

AU - Spijkers, L J A

AU - Lemkens, P

AU - Nelissen, J

AU - Compeer, M G

AU - Alewijnse, A E

AU - De Mey, J G R

N1 - © 2011 The Authors. British Journal of Pharmacology © 2011 The British Pharmacological Society.

PY - 2012

Y1 - 2012

N2 - BACKGROUND AND PURPOSE: Calcitonin gene-related peptide (CGRP) has been proposed to relax vascular smooth muscle cells (VSMC) via cAMP and can promote dissociation of endothelin-1 (ET-1) from ET(A) receptors. The latter is not mimicked by other stimuli of adenylate cyclases. Therefore, we evaluated the involvement of G-protein βγ subunits (Gβγ) in the arterial effects of CGRP receptor stimulation.EXPERIMENTAL APPROACH: To test the hypothesis that instead of α subunits of G-proteins (Gαs), Gβγ mediates the effects of CGRP receptor activation, we used (i) rat isolated mesenteric resistance arteries (MRA), (ii) pharmacological modulators of cyclic nucleotides; and (iii) low molecular weight inhibitors of the functions of Gβγ, gallein and M119. To validate these tools with respect to CGRP receptor function, we performed organ bath studies with rat isolated MRA, radioligand binding on membranes from CHO cells expressing human CGRP receptors and cAMP production assays in rat cultured VSMC.KEY RESULTS: In isolated arteries contracted with K(+) or ET-1, IBMX (PDE inhibitor) increased sodium nitroprusside (SNP)- and isoprenaline (ISO)- but not CGRP-induced relaxations. While fluorescein (negative control) was without effects, gallein increased binding of [(125) I]-CGRP in the absence and presence of GTPγS. Gallein also increased CGRP-induced cAMP production in VSMC. Despite these stimulating effects, gallein and M119 selectively inhibited the relaxing and anti-endothelinergic effects of CGRP in isolated arteries while not altering contractile responses to K(+) or ET-1 or relaxing responses to ISO or SNP.CONCLUSION AND IMPLICATIONS: Activated CGRP receptors induce cyclic nucleotide-independent relaxation of VSMC and terminate arterial effects of ET-1 via Gβγ.

AB - BACKGROUND AND PURPOSE: Calcitonin gene-related peptide (CGRP) has been proposed to relax vascular smooth muscle cells (VSMC) via cAMP and can promote dissociation of endothelin-1 (ET-1) from ET(A) receptors. The latter is not mimicked by other stimuli of adenylate cyclases. Therefore, we evaluated the involvement of G-protein βγ subunits (Gβγ) in the arterial effects of CGRP receptor stimulation.EXPERIMENTAL APPROACH: To test the hypothesis that instead of α subunits of G-proteins (Gαs), Gβγ mediates the effects of CGRP receptor activation, we used (i) rat isolated mesenteric resistance arteries (MRA), (ii) pharmacological modulators of cyclic nucleotides; and (iii) low molecular weight inhibitors of the functions of Gβγ, gallein and M119. To validate these tools with respect to CGRP receptor function, we performed organ bath studies with rat isolated MRA, radioligand binding on membranes from CHO cells expressing human CGRP receptors and cAMP production assays in rat cultured VSMC.KEY RESULTS: In isolated arteries contracted with K(+) or ET-1, IBMX (PDE inhibitor) increased sodium nitroprusside (SNP)- and isoprenaline (ISO)- but not CGRP-induced relaxations. While fluorescein (negative control) was without effects, gallein increased binding of [(125) I]-CGRP in the absence and presence of GTPγS. Gallein also increased CGRP-induced cAMP production in VSMC. Despite these stimulating effects, gallein and M119 selectively inhibited the relaxing and anti-endothelinergic effects of CGRP in isolated arteries while not altering contractile responses to K(+) or ET-1 or relaxing responses to ISO or SNP.CONCLUSION AND IMPLICATIONS: Activated CGRP receptors induce cyclic nucleotide-independent relaxation of VSMC and terminate arterial effects of ET-1 via Gβγ.

KW - 1-Methyl-3-isobutylxanthine

KW - Animals

KW - CHO Cells

KW - Calcitonin Gene-Related Peptide

KW - Cricetinae

KW - Cricetulus

KW - Cyclic AMP

KW - Cyclohexanes

KW - GTP-Binding Protein beta Subunits

KW - GTP-Binding Protein gamma Subunits

KW - Humans

KW - Isoproterenol

KW - Male

KW - Mesenteric Arteries

KW - Muscle, Smooth, Vascular

KW - Nitroprusside

KW - Rats

KW - Rats, Inbred WKY

KW - Receptors, Calcitonin Gene-Related Peptide

KW - Vasodilation

KW - Xanthenes

U2 - 10.1111/j.1476-5381.2011.01774.x

DO - 10.1111/j.1476-5381.2011.01774.x

M3 - Journal article

VL - 166

SP - 297

EP - 308

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 1

ER -