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Article: Morphological heterogeneity with normal expression but altered function of G proteins in porcine cultured regenerated coronary endothelial cells

TitleMorphological heterogeneity with normal expression but altered function of G proteins in porcine cultured regenerated coronary endothelial cells
Authors
Issue Date1997
PublisherJohn Wiley & Sons Ltd. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=0007-1188&site=1
Citation
British Journal Of Pharmacology, 1997, v. 122 n. 6, p. 999-1008 How to Cite?
Abstract1. Experiments were designed to investigate whether the pertussis toxin-dependent endothelial dysfunction following balloon injury is due to a reduced expression or an insufficient function of G-proteins. 2. Endothelium-dependent responses of porcine coronary arteries were examined in vitro by use of conventional organ chambers. Morphological analysis was performed by isolating and culturing the endothelial cells from these arteries. The expression of Gi-proteins in regenerated endothelial cells was measured by Western blots and immunolabelling. The function of G-proteins was assessed by measuring the GTPase activity of cultured endothelial cells. 3. Eight days following denudation, endothelial regrowth was confirmed by histological examination and by demonstrating presence of endothelium-dependent relaxations to bradykinin and 5-hydroxytryptamine (5-HT). In primary culture, the regenerated endothelial cells displayed a 'cobblestone' pattern as seen with native endothelial cells. 4. Twenty eight days after denudation, the endothelium-dependent relaxations induced by 5-HT were impaired, but those to bradykinin were maintained. However, the latter were reduced when endothelium dependent hyperpolarization was prevented. 5. Twenty eight days after denudation, multinucleated giant cells were present in the regenerated but not in the native cultured endothelial cell populations. These regenerated endothelial cells incorporated less tritiated thymidine than native endothelial cells. 6. The intensities of the bands on the immunoblot of the regenerated endothelial cells, when several antibodies against Giα1/α2/α3 were used, were the same as those obtained in native endothelial cells. The immunolabelling with the same antibodies was similar between the giant cells and the regenerated endothelial cells of normal size. The hydrolysis of GTP was lower in regenerated than in native endothelial cell membranes. 7. In conclusion, endothelium-dependent relaxations mediated by Gi-proteins are impaired in balloon denuded coronary arteries. This dysfunction following regeneration cannot be explained by a reduced expression of Gi proteins but rather reflects an abnormal function of the G-proteins in the regenerated endothelium.
Persistent Identifierhttp://hdl.handle.net/10722/171198
ISSN
2015 Impact Factor: 5.259
2015 SCImago Journal Rankings: 2.368
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorBorgCapra, Cen_US
dc.contributor.authorFournetBourguignon, MPen_US
dc.contributor.authorJaniak, Pen_US
dc.contributor.authorVilleneuve, Nen_US
dc.contributor.authorBidouard, JPen_US
dc.contributor.authorVilaine, JPen_US
dc.contributor.authorVanhoutte, PMen_US
dc.date.accessioned2012-10-30T06:12:39Z-
dc.date.available2012-10-30T06:12:39Z-
dc.date.issued1997en_US
dc.identifier.citationBritish Journal Of Pharmacology, 1997, v. 122 n. 6, p. 999-1008en_US
dc.identifier.issn0007-1188en_US
dc.identifier.urihttp://hdl.handle.net/10722/171198-
dc.description.abstract1. Experiments were designed to investigate whether the pertussis toxin-dependent endothelial dysfunction following balloon injury is due to a reduced expression or an insufficient function of G-proteins. 2. Endothelium-dependent responses of porcine coronary arteries were examined in vitro by use of conventional organ chambers. Morphological analysis was performed by isolating and culturing the endothelial cells from these arteries. The expression of Gi-proteins in regenerated endothelial cells was measured by Western blots and immunolabelling. The function of G-proteins was assessed by measuring the GTPase activity of cultured endothelial cells. 3. Eight days following denudation, endothelial regrowth was confirmed by histological examination and by demonstrating presence of endothelium-dependent relaxations to bradykinin and 5-hydroxytryptamine (5-HT). In primary culture, the regenerated endothelial cells displayed a 'cobblestone' pattern as seen with native endothelial cells. 4. Twenty eight days after denudation, the endothelium-dependent relaxations induced by 5-HT were impaired, but those to bradykinin were maintained. However, the latter were reduced when endothelium dependent hyperpolarization was prevented. 5. Twenty eight days after denudation, multinucleated giant cells were present in the regenerated but not in the native cultured endothelial cell populations. These regenerated endothelial cells incorporated less tritiated thymidine than native endothelial cells. 6. The intensities of the bands on the immunoblot of the regenerated endothelial cells, when several antibodies against Giα1/α2/α3 were used, were the same as those obtained in native endothelial cells. The immunolabelling with the same antibodies was similar between the giant cells and the regenerated endothelial cells of normal size. The hydrolysis of GTP was lower in regenerated than in native endothelial cell membranes. 7. In conclusion, endothelium-dependent relaxations mediated by Gi-proteins are impaired in balloon denuded coronary arteries. This dysfunction following regeneration cannot be explained by a reduced expression of Gi proteins but rather reflects an abnormal function of the G-proteins in the regenerated endothelium.en_US
dc.languageengen_US
dc.publisherJohn Wiley & Sons Ltd. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=0007-1188&site=1en_US
dc.relation.ispartofBritish Journal of Pharmacologyen_US
dc.subject.meshAnimalsen_US
dc.subject.meshArteries - Enzymology - Metabolism - Physiologyen_US
dc.subject.meshBlotting, Westernen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshCoronary Vessels - Enzymology - Metabolism - Physiologyen_US
dc.subject.meshDna Replicationen_US
dc.subject.meshFemaleen_US
dc.subject.meshFluorescent Antibody Technique, Indirecten_US
dc.subject.meshGtp Phosphohydrolases - Metabolismen_US
dc.subject.meshGtp-Binding Proteins - Metabolismen_US
dc.subject.meshMaleen_US
dc.subject.meshOrgan Culture Techniquesen_US
dc.subject.meshRegenerationen_US
dc.subject.meshSwineen_US
dc.titleMorphological heterogeneity with normal expression but altered function of G proteins in porcine cultured regenerated coronary endothelial cellsen_US
dc.typeArticleen_US
dc.identifier.emailVanhoutte, PM:vanhoutt@hku.hken_US
dc.identifier.authorityVanhoutte, PM=rp00238en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1038/sj.bjp.0701475en_US
dc.identifier.pmid9401761-
dc.identifier.scopuseid_2-s2.0-0030786712en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0030786712&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume122en_US
dc.identifier.issue6en_US
dc.identifier.spage999en_US
dc.identifier.epage1008en_US
dc.identifier.isiWOS:A1997YG22000005-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridBorgCapra, C=24473996400en_US
dc.identifier.scopusauthoridFournetBourguignon, MP=6507287770en_US
dc.identifier.scopusauthoridJaniak, P=6603686655en_US
dc.identifier.scopusauthoridVilleneuve, N=7003458215en_US
dc.identifier.scopusauthoridBidouard, JP=6601955808en_US
dc.identifier.scopusauthoridVilaine, JP=7004617134en_US
dc.identifier.scopusauthoridVanhoutte, PM=7202304247en_US

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