Functional characterization and expression of endothelin receptors in rat carotid artery: involvement of nitric oxide, a vasodilator prostanoid and the opening of K+ channels in ETB‐induced relaxation
We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ETA and ETB receptors, evaluated by reverse transcription–polymerase chain reaction (RT–PCR) and Western immunoblotting, were detected in carotid segments. Immunohistochemi...
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| Published in | British journal of pharmacology Vol. 146; no. 6; pp. 903 - 912 |
|---|---|
| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Oxford, UK
Blackwell Publishing Ltd
01.11.2005
Nature Publishing |
| Subjects | |
| Online Access | Get full text |
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| Abstract | We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ETA and ETB receptors, evaluated by reverse transcription–polymerase chain reaction (RT–PCR) and Western immunoblotting, were detected in carotid segments. Immunohistochemical assays showed that ETB receptors are expressed in the endothelium and smooth muscle cells, while ETA receptors are expressed only in the smooth muscle cells. In endothelium‐denuded vessels, levels of ETB receptor mRNA were reduced.
Vascular reactivity experiments, using standard muscle bath procedures, showed that ET‐1 induces contraction in endothelium‐intact and ‐denuded carotid rings in a concentration‐dependent manner. Endothelial removal enhanced ET‐1‐induced contraction. BQ123 and BQ788, selective antagonists for ETA and ETB receptors, respectively, produced concentration‐dependent rightward displacements of the ET‐1 concentration–response curves.
IRL1620, a selective agonist for ETB receptors, induced a slight vasoconstriction that was abolished by BQ788, but not affected by BQ123. IRL1620‐induced contraction was augmented after endothelium removal.
ET‐1 concentration dependently relaxed phenylephrine‐precontracted rings with intact endothelium. The relaxation was augmented in the presence of BQ123, reduced in the presence of BQ788 and completely abolished after endothelium removal. IRL1620 induced vasorelaxation that was abolished by BQ788 and endothelium removal, but not affected by BQ123.
Preincubation of intact rings with NG‐nitro‐L‐arginine methyl ester (L‐NAME), 1H‐[1,2,4]oxadiazolo[4,3‐a]quinoxalin‐1‐one (ODQ), indomethacin or tetraethylammonium (TEA) reduced IRL1620‐induced relaxation. The combination of L‐NAME, indomethacin and TEA completely abolished IRL1620‐induced relaxation while sulfaphenazole did not affect this response. 4‐aminopyridine (4‐AP), but not apamin, glibenclamide or charybdotoxin, reduced IRL1620‐induced relaxation.
The major finding of this work is that it firstly demonstrated functionally the existence of both ETA and ETB vasoconstrictor receptors located on the smooth muscle of rat carotid arteries and endothelial ETB receptors that mediated vasorelaxation via NO–cGMP pathway, vasodilator cyclooxygenase product(s) and the activation of voltage‐dependent K+ channels.
British Journal of Pharmacology (2005) 146, 903–912. doi:10.1038/sj.bjp.0706388 |
|---|---|
| AbstractList | We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ETA and ETB receptors, evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western immunoblotting, were detected in carotid segments. Immunohistochemical assays showed that ETB receptors are expressed in the endothelium and smooth muscle cells, while ETA receptors are expressed only in the smooth muscle cells. In endothelium-denuded vessels, levels of ETB receptor mRNA were reduced. Vascular reactivity experiments, using standard muscle bath procedures, showed that ET-1 induces contraction in endothelium-intact and -denuded carotid rings in a concentration-dependent manner. Endothelial removal enhanced ET-1-induced contraction. BQ123 and BQ788, selective antagonists for ETA and ETB receptors, respectively, produced concentration-dependent rightward displacements of the ET-1 concentration-response curves. IRL1620, a selective agonist for ETB receptors, induced a slight vasoconstriction that was abolished by BQ788, but not affected by BQ123. IRL1620-induced contraction was augmented after endothelium removal. ET-1 concentration dependently relaxed phenylephrine-precontracted rings with intact endothelium. The relaxation was augmented in the presence of BQ123, reduced in the presence of BQ788 and completely abolished after endothelium removal. IRL1620 induced vasorelaxation that was abolished by BQ788 and endothelium removal, but not affected by BQ123. Preincubation of intact rings with N(G)-nitro-L-arginine methyl ester (L-NAME), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), indomethacin or tetraethylammonium (TEA) reduced IRL1620-induced relaxation. The combination of L-NAME, indomethacin and TEA completely abolished IRL1620-induced relaxation while sulfaphenazole did not affect this response. 4-aminopyridine (4-AP), but not apamin, glibenclamide or charybdotoxin, reduced IRL1620-induced relaxation. The major finding of this work is that it firstly demonstrated functionally the existence of both ETA and ETB vasoconstrictor receptors located on the smooth muscle of rat carotid arteries and endothelial ETB receptors that mediated vasorelaxation via NO-cGMP pathway, vasodilator cyclooxygenase product(s) and the activation of voltage-dependent K+ channels.We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ETA and ETB receptors, evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western immunoblotting, were detected in carotid segments. Immunohistochemical assays showed that ETB receptors are expressed in the endothelium and smooth muscle cells, while ETA receptors are expressed only in the smooth muscle cells. In endothelium-denuded vessels, levels of ETB receptor mRNA were reduced. Vascular reactivity experiments, using standard muscle bath procedures, showed that ET-1 induces contraction in endothelium-intact and -denuded carotid rings in a concentration-dependent manner. Endothelial removal enhanced ET-1-induced contraction. BQ123 and BQ788, selective antagonists for ETA and ETB receptors, respectively, produced concentration-dependent rightward displacements of the ET-1 concentration-response curves. IRL1620, a selective agonist for ETB receptors, induced a slight vasoconstriction that was abolished by BQ788, but not affected by BQ123. IRL1620-induced contraction was augmented after endothelium removal. ET-1 concentration dependently relaxed phenylephrine-precontracted rings with intact endothelium. The relaxation was augmented in the presence of BQ123, reduced in the presence of BQ788 and completely abolished after endothelium removal. IRL1620 induced vasorelaxation that was abolished by BQ788 and endothelium removal, but not affected by BQ123. Preincubation of intact rings with N(G)-nitro-L-arginine methyl ester (L-NAME), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), indomethacin or tetraethylammonium (TEA) reduced IRL1620-induced relaxation. The combination of L-NAME, indomethacin and TEA completely abolished IRL1620-induced relaxation while sulfaphenazole did not affect this response. 4-aminopyridine (4-AP), but not apamin, glibenclamide or charybdotoxin, reduced IRL1620-induced relaxation. The major finding of this work is that it firstly demonstrated functionally the existence of both ETA and ETB vasoconstrictor receptors located on the smooth muscle of rat carotid arteries and endothelial ETB receptors that mediated vasorelaxation via NO-cGMP pathway, vasodilator cyclooxygenase product(s) and the activation of voltage-dependent K+ channels. We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ET A and ET B receptors, evaluated by reverse transcription–polymerase chain reaction (RT–PCR) and Western immunoblotting, were detected in carotid segments. Immunohistochemical assays showed that ET B receptors are expressed in the endothelium and smooth muscle cells, while ET A receptors are expressed only in the smooth muscle cells. In endothelium-denuded vessels, levels of ET B receptor mRNA were reduced. Vascular reactivity experiments, using standard muscle bath procedures, showed that ET-1 induces contraction in endothelium-intact and -denuded carotid rings in a concentration-dependent manner. Endothelial removal enhanced ET-1-induced contraction. BQ123 and BQ788, selective antagonists for ET A and ET B receptors, respectively, produced concentration-dependent rightward displacements of the ET-1 concentration–response curves. IRL1620, a selective agonist for ET B receptors, induced a slight vasoconstriction that was abolished by BQ788, but not affected by BQ123. IRL1620-induced contraction was augmented after endothelium removal. ET-1 concentration dependently relaxed phenylephrine-precontracted rings with intact endothelium. The relaxation was augmented in the presence of BQ123, reduced in the presence of BQ788 and completely abolished after endothelium removal. IRL1620 induced vasorelaxation that was abolished by BQ788 and endothelium removal, but not affected by BQ123. Preincubation of intact rings with N G -nitro- L -arginine methyl ester ( L -NAME), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), indomethacin or tetraethylammonium (TEA) reduced IRL1620-induced relaxation. The combination of L -NAME, indomethacin and TEA completely abolished IRL1620-induced relaxation while sulfaphenazole did not affect this response. 4-aminopyridine (4-AP), but not apamin, glibenclamide or charybdotoxin, reduced IRL1620-induced relaxation. The major finding of this work is that it firstly demonstrated functionally the existence of both ET A and ET B vasoconstrictor receptors located on the smooth muscle of rat carotid arteries and endothelial ET B receptors that mediated vasorelaxation via NO–cGMP pathway, vasodilator cyclooxygenase product(s) and the activation of voltage-dependent K + channels. We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ETA and ETB receptors, evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western immunoblotting, were detected in carotid segments. Immunohistochemical assays showed that ETB receptors are expressed in the endothelium and smooth muscle cells, while ETA receptors are expressed only in the smooth muscle cells. In endothelium-denuded vessels, levels of ETB receptor mRNA were reduced. Vascular reactivity experiments, using standard muscle bath procedures, showed that ET-1 induces contraction in endothelium-intact and -denuded carotid rings in a concentration-dependent manner. Endothelial removal enhanced ET-1-induced contraction. BQ123 and BQ788, selective antagonists for ETA and ETB receptors, respectively, produced concentration-dependent rightward displacements of the ET-1 concentration-response curves. IRL1620, a selective agonist for ETB receptors, induced a slight vasoconstriction that was abolished by BQ788, but not affected by BQ123. IRL1620-induced contraction was augmented after endothelium removal. ET-1 concentration dependently relaxed phenylephrine-precontracted rings with intact endothelium. The relaxation was augmented in the presence of BQ123, reduced in the presence of BQ788 and completely abolished after endothelium removal. IRL1620 induced vasorelaxation that was abolished by BQ788 and endothelium removal, but not affected by BQ123. Preincubation of intact rings with N(G)-nitro-L-arginine methyl ester (L-NAME), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), indomethacin or tetraethylammonium (TEA) reduced IRL1620-induced relaxation. The combination of L-NAME, indomethacin and TEA completely abolished IRL1620-induced relaxation while sulfaphenazole did not affect this response. 4-aminopyridine (4-AP), but not apamin, glibenclamide or charybdotoxin, reduced IRL1620-induced relaxation. The major finding of this work is that it firstly demonstrated functionally the existence of both ETA and ETB vasoconstrictor receptors located on the smooth muscle of rat carotid arteries and endothelial ETB receptors that mediated vasorelaxation via NO-cGMP pathway, vasodilator cyclooxygenase product(s) and the activation of voltage-dependent K+ channels. We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ETA and ETB receptors, evaluated by reverse transcription–polymerase chain reaction (RT–PCR) and Western immunoblotting, were detected in carotid segments. Immunohistochemical assays showed that ETB receptors are expressed in the endothelium and smooth muscle cells, while ETA receptors are expressed only in the smooth muscle cells. In endothelium‐denuded vessels, levels of ETB receptor mRNA were reduced. Vascular reactivity experiments, using standard muscle bath procedures, showed that ET‐1 induces contraction in endothelium‐intact and ‐denuded carotid rings in a concentration‐dependent manner. Endothelial removal enhanced ET‐1‐induced contraction. BQ123 and BQ788, selective antagonists for ETA and ETB receptors, respectively, produced concentration‐dependent rightward displacements of the ET‐1 concentration–response curves. IRL1620, a selective agonist for ETB receptors, induced a slight vasoconstriction that was abolished by BQ788, but not affected by BQ123. IRL1620‐induced contraction was augmented after endothelium removal. ET‐1 concentration dependently relaxed phenylephrine‐precontracted rings with intact endothelium. The relaxation was augmented in the presence of BQ123, reduced in the presence of BQ788 and completely abolished after endothelium removal. IRL1620 induced vasorelaxation that was abolished by BQ788 and endothelium removal, but not affected by BQ123. Preincubation of intact rings with NG‐nitro‐L‐arginine methyl ester (L‐NAME), 1H‐[1,2,4]oxadiazolo[4,3‐a]quinoxalin‐1‐one (ODQ), indomethacin or tetraethylammonium (TEA) reduced IRL1620‐induced relaxation. The combination of L‐NAME, indomethacin and TEA completely abolished IRL1620‐induced relaxation while sulfaphenazole did not affect this response. 4‐aminopyridine (4‐AP), but not apamin, glibenclamide or charybdotoxin, reduced IRL1620‐induced relaxation. The major finding of this work is that it firstly demonstrated functionally the existence of both ETA and ETB vasoconstrictor receptors located on the smooth muscle of rat carotid arteries and endothelial ETB receptors that mediated vasorelaxation via NO–cGMP pathway, vasodilator cyclooxygenase product(s) and the activation of voltage‐dependent K+ channels. British Journal of Pharmacology (2005) 146, 903–912. doi:10.1038/sj.bjp.0706388 |
| Author | De Oliveira, Ana M Tirapelli, Carlos R Yogi, Alvaro Tostes, Rita C Legros, Eurode Casolari, Debora A Montezano, Augusto C D'Orléans‐Juste, Pedro |
| AuthorAffiliation | 1 Department of Pharmacology, Faculty of Medicine of Ribeirão Preto, University of São Paulo (USP), SP, Brazil 2 Department of Pharmacology, Institute of Biomedical Sciences, USP, São Paulo, SP, Brazil 4 Department of Physics and Chemistry, Laboratory of Pharmacology, Faculty of Pharmaceutical Sciences, USP, Ribeirão Preto, SP, Brazil 3 Department of Pharmacology, Medical School, Institut de pharmacologie de Sherbrooke, Universite de Sherbrooke, QC, Canada |
| AuthorAffiliation_xml | – name: 3 Department of Pharmacology, Medical School, Institut de pharmacologie de Sherbrooke, Universite de Sherbrooke, QC, Canada – name: 4 Department of Physics and Chemistry, Laboratory of Pharmacology, Faculty of Pharmaceutical Sciences, USP, Ribeirão Preto, SP, Brazil – name: 2 Department of Pharmacology, Institute of Biomedical Sciences, USP, São Paulo, SP, Brazil – name: 1 Department of Pharmacology, Faculty of Medicine of Ribeirão Preto, University of São Paulo (USP), SP, Brazil |
| Author_xml | – sequence: 1 givenname: Carlos R surname: Tirapelli fullname: Tirapelli, Carlos R – sequence: 2 givenname: Debora A surname: Casolari fullname: Casolari, Debora A – sequence: 3 givenname: Alvaro surname: Yogi fullname: Yogi, Alvaro – sequence: 4 givenname: Augusto C surname: Montezano fullname: Montezano, Augusto C – sequence: 5 givenname: Rita C surname: Tostes fullname: Tostes, Rita C – sequence: 6 givenname: Eurode surname: Legros fullname: Legros, Eurode – sequence: 7 givenname: Pedro surname: D'Orléans‐Juste fullname: D'Orléans‐Juste, Pedro – sequence: 8 givenname: Ana M surname: De Oliveira fullname: De Oliveira, Ana M |
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| Keywords | Vasodilator agent Rat Rodentia relaxation Ionic channel Endothelin 1 Inorganic element Artery Characterization ETB receptor Vertebrata Mammalia Blood vessel Animal Nitric oxide ETB endothelin receptor Prostanoid Circulatory system Carotid artery endothelin-1 Potassium Endothelin receptor |
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| Snippet | We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ETA and ETB receptors,... We aimed to functionally characterize endothelin (ET) receptors in the rat carotid artery. mRNA and protein expressions of both ET A and ET B receptors,... |
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| SubjectTerms | Animals Biological and medical sciences Blotting, Western Carotid Arteries - drug effects Carotid Arteries - metabolism Carotid Arteries - physiology Carotid artery Endothelin A Receptor Antagonists Endothelin B Receptor Antagonists Endothelin-1 - pharmacology Endothelins - pharmacology endothelin‐1 Endothelium, Vascular - physiology Endothelium-Dependent Relaxing Factors - physiology ETB receptor Gene Expression - drug effects Immunohistochemistry Male Medical sciences Nitric Oxide - physiology Oligopeptides - pharmacology Peptide Fragments - pharmacology Peptides, Cyclic - pharmacology Pharmacology. Drug treatments Piperidines - pharmacology Potassium Channels - physiology Rats Rats, Wistar Receptor, Endothelin A - genetics Receptor, Endothelin A - physiology Receptor, Endothelin B - genetics Receptor, Endothelin B - physiology relaxation Reverse Transcriptase Polymerase Chain Reaction - methods RNA, Messenger - genetics RNA, Messenger - metabolism Vasodilation - drug effects Vasodilation - physiology |
| Title | Functional characterization and expression of endothelin receptors in rat carotid artery: involvement of nitric oxide, a vasodilator prostanoid and the opening of K+ channels in ETB‐induced relaxation |
| URI | https://onlinelibrary.wiley.com/doi/abs/10.1038%2Fsj.bjp.0706388 https://www.ncbi.nlm.nih.gov/pubmed/16151434 https://www.proquest.com/docview/217205214 https://www.proquest.com/docview/69062682 https://pubmed.ncbi.nlm.nih.gov/PMC1751214 |
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