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Article: β-adrenoceptor-mediated down-regulation of M2 muscarinic receptors: Role of cyclic adenosine 5′-monophosphate-dependent protein kinase and protein kinase C

Titleβ-adrenoceptor-mediated down-regulation of M2 muscarinic receptors: Role of cyclic adenosine 5′-monophosphate-dependent protein kinase and protein kinase C
Authors
Issue Date1996
PublisherAmerican Society for Pharmacology and Experimental Therapeutics. The Journal's web site is located at http://www.molpharm.org
Citation
Molecular Pharmacology, 1996, v. 49 n. 4, p. 629-635 How to Cite?
AbstractStimulation of β2-adrenoceptors with the selective β2 agonist procaterol caused a biphasic decrease in cell surface M2 muscarinic receptor number in human embryonic lung 299 cells when measured with the hydrophilic antagonist [3H]N-methylscopolamine. In contrast, total muscarinic receptor number, measured with the lipophilic antagonist [3H]quinuclidinylbenzilate, decreased after only 24-hr treatments with procaterol. The loss in receptor number at 24 hr was mimicked with the use of forskolin and the cAMP analogue 8-bromo-cAMP, indicating a cAMP-mediated mechanism. Northern blot analysis showed a small and transient increase in m2-receptor mRNA levels up to 2 hr but no long term (24 hr) effect. Chronic (24-hr) treatment with 8-bromo-cAMP also had no effect on m2 muscarinic receptor mRNA, whereas forskolin caused a 50% reduction in the steady state levels of m2 mRNA that could be only partially blocked by the cAMP-dependent protein kinase inhibitor H-8 and the protein kinase C inhibitor GF 109203X. Procaterol-induced down-regulation of M2 receptors was fully blocked by N-[2-(methylamino)ethyl]-5′-isoquinoline-sulfonamide and 2-[1-(3-dimethylaminopropyl)-inol-3-yl]-3-(indol-3-yl)maleimide, implicating both of these kinases in the M2 muscarinic receptor down-regulation. Conversely, the forskolin- and 8-bromo-cAMP-induced down-regulation was only partially inhibited and unaffected by these inhibitors, respectively. In control cells and those treated with procaterol for ≤2 hr, cAMP generation was significantly inhibited by carbachol. The inhibitory effect of carbachol was, however, lost after 24-hr exposure to procaterol. This desensitization was partially reversed by preincubations with H-8 and GF 109203X. Collectively, these results suggest that transregulation of M2 muscarinic receptors by β2-adrenoceptor stimulation can be demonstrated at the protein level in human embryonic lung 299 cells. Furthermore, a role is suggested for cAMP-dependent kinase and PKC in M2 muscarinic receptor down-regulation and their functional desensitization.
Persistent Identifierhttp://hdl.handle.net/10722/162115
ISSN
2015 Impact Factor: 3.931
2015 SCImago Journal Rankings: 2.047
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorRousell, Jen_US
dc.contributor.authorHaddad, EBen_US
dc.contributor.authorMak, JCWen_US
dc.contributor.authorWebb, BLJen_US
dc.contributor.authorGiembycz, MAen_US
dc.contributor.authorBarnes, PJen_US
dc.date.accessioned2012-09-05T05:17:25Z-
dc.date.available2012-09-05T05:17:25Z-
dc.date.issued1996en_US
dc.identifier.citationMolecular Pharmacology, 1996, v. 49 n. 4, p. 629-635en_US
dc.identifier.issn0026-895Xen_US
dc.identifier.urihttp://hdl.handle.net/10722/162115-
dc.description.abstractStimulation of β2-adrenoceptors with the selective β2 agonist procaterol caused a biphasic decrease in cell surface M2 muscarinic receptor number in human embryonic lung 299 cells when measured with the hydrophilic antagonist [3H]N-methylscopolamine. In contrast, total muscarinic receptor number, measured with the lipophilic antagonist [3H]quinuclidinylbenzilate, decreased after only 24-hr treatments with procaterol. The loss in receptor number at 24 hr was mimicked with the use of forskolin and the cAMP analogue 8-bromo-cAMP, indicating a cAMP-mediated mechanism. Northern blot analysis showed a small and transient increase in m2-receptor mRNA levels up to 2 hr but no long term (24 hr) effect. Chronic (24-hr) treatment with 8-bromo-cAMP also had no effect on m2 muscarinic receptor mRNA, whereas forskolin caused a 50% reduction in the steady state levels of m2 mRNA that could be only partially blocked by the cAMP-dependent protein kinase inhibitor H-8 and the protein kinase C inhibitor GF 109203X. Procaterol-induced down-regulation of M2 receptors was fully blocked by N-[2-(methylamino)ethyl]-5′-isoquinoline-sulfonamide and 2-[1-(3-dimethylaminopropyl)-inol-3-yl]-3-(indol-3-yl)maleimide, implicating both of these kinases in the M2 muscarinic receptor down-regulation. Conversely, the forskolin- and 8-bromo-cAMP-induced down-regulation was only partially inhibited and unaffected by these inhibitors, respectively. In control cells and those treated with procaterol for ≤2 hr, cAMP generation was significantly inhibited by carbachol. The inhibitory effect of carbachol was, however, lost after 24-hr exposure to procaterol. This desensitization was partially reversed by preincubations with H-8 and GF 109203X. Collectively, these results suggest that transregulation of M2 muscarinic receptors by β2-adrenoceptor stimulation can be demonstrated at the protein level in human embryonic lung 299 cells. Furthermore, a role is suggested for cAMP-dependent kinase and PKC in M2 muscarinic receptor down-regulation and their functional desensitization.en_US
dc.languageengen_US
dc.publisherAmerican Society for Pharmacology and Experimental Therapeutics. The Journal's web site is located at http://www.molpharm.orgen_US
dc.relation.ispartofMolecular Pharmacologyen_US
dc.subject.meshCyclic Amp-Dependent Protein Kinases - Physiologyen_US
dc.subject.meshDown-Regulationen_US
dc.subject.meshHumansen_US
dc.subject.meshProtein Kinase C - Physiologyen_US
dc.subject.meshRna, Messenger - Analysisen_US
dc.subject.meshRadioligand Assayen_US
dc.subject.meshReceptors, Adrenergic, Beta-2 - Physiologyen_US
dc.subject.meshReceptors, Muscarinic - Analysis - Geneticsen_US
dc.titleβ-adrenoceptor-mediated down-regulation of M2 muscarinic receptors: Role of cyclic adenosine 5′-monophosphate-dependent protein kinase and protein kinase Cen_US
dc.typeArticleen_US
dc.identifier.emailMak, JCW:judymak@hku.hken_US
dc.identifier.authorityMak, JCW=rp00352en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid8609890-
dc.identifier.scopuseid_2-s2.0-0029664507en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0029664507&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume49en_US
dc.identifier.issue4en_US
dc.identifier.spage629en_US
dc.identifier.epage635en_US
dc.identifier.isiWOS:A1996UD68900007-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridRousell, J=6602560061en_US
dc.identifier.scopusauthoridHaddad, EB=7102803008en_US
dc.identifier.scopusauthoridMak, JCW=7103323094en_US
dc.identifier.scopusauthoridWebb, BLJ=37017893400en_US
dc.identifier.scopusauthoridGiembycz, MA=7007035171en_US
dc.identifier.scopusauthoridBarnes, PJ=36064679400en_US

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