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Stimulatory activity of anti-peptide antibodies against the second extracellular loop of human M2 muscarinic receptors.

Artikel i vetenskaplig tidskrift
Författare W Wang
G Guo
J Tang
J Li
R Zhao
Åke Hjalmarson
Michael Fu
Publicerad i Chinese medical journal
Volym 113
Nummer/häfte 10
Sidor 867-71
ISSN 0366-6999
Publiceringsår 2000
Publicerad vid Wallenberglaboratoriet
Sidor 867-71
Språk en
Länkar www.ncbi.nlm.nih.gov/entrez/query.f...
Ämnesord Amino Acid Sequence, Animals, Antibodies, immunology, Atropine, pharmacology, Calcium, metabolism, Carbachol, pharmacology, Cyclic AMP, biosynthesis, Female, Guinea Pigs, Humans, Male, Molecular Sequence Data, Peptide Fragments, immunology, Receptor, Muscarinic M2, Receptors, Muscarinic, immunology
Ämneskategorier Fysiologi, Cell- och molekylärbiologi, Mikrobiologi inom det medicinska området

Sammanfattning

OBJECTIVE: To study the activity of anti-peptide antibodies against the second extracellular loop of human M2 muscarinic receptors on cAMP production and inward calcium currents (Ica) in guinea pig ventricular myocytes. A comparison was also made with those of a muscarinic receptor agonist. METHODS: cAMP content was determined by radioimmunoassay and the Ica in guinea pig single ventricular cells were recorded by the whole-cell patch clamp technique. RESULTS: Both the muscarinic receptor agonist, carbachol (Carb 10 mumol/L), and anti-peptide antibodies (Abs 100 nmol/L) could decrease basal cAMP levels (by 46.9% +/- 4.2% and 60.2% +/- 4.6%, respectively) and basal Ica. Both Carb (10 mumol/L) and Abs (100 nmol/L) could also inhibit the isoprenaline-induced (Iso 0.8 mumol/L) increases in cAMP production (from 108.2 +/- 7.0 to 88.4 +/- 7.2 pmol/mg.protein/min for Carb and 88.6 +/- 5.1 pmol/mg.protein/min for Abs, respectively) and the increases in Ica. The muscarinic receptor antagonist atropine (Atr) was able to prevent these effects of Carb and Abs. CONCLUSIONS: Anti-peptide antibodies against an epitope located in the second extracellular loop of human M2 muscarinic receptors, similar to muscarinic receptor agonist, could decrease the basal Ica and beta-receptor agonist stimulated increase of Ica by decreasing the basal and beta-receptor agonist stimulated increase of cAMP production, and therefore could have an effect on their target receptor. These results further suggest that autoimmunity may participate in the pathogenesis of human cardiomyopathy and the second extracellular loop of human M2 muscarinic receptor could be the main immunodominant region.

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