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Immunocytochemical localization of M2 muscarinic receptors in rat ventricles with anti-peptide antibodies.

Artikel i vetenskaplig tidskrift
Författare Michael Fu
W Schulze
W P Wolf
Åke Hjalmarson
Johan Hoebeke
Publicerad i The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
Volym 42
Nummer/häfte 3
Sidor 337-43
ISSN 0022-1554
Publiceringsår 1994
Publicerad vid Wallenberglaboratoriet
Sidor 337-43
Språk en
Länkar www.ncbi.nlm.nih.gov/entrez/query.f...
Ämnesord Acetylcholine, chemistry, Amino Acid Sequence, Animals, Antibodies, Fluorescent Antibody Technique, Heart Ventricles, chemistry, ultrastructure, Humans, Immunoblotting, Immunohistochemistry, Microscopy, Immunoelectron, methods, Molecular Sequence Data, Peptides, Rabbits, Rats, Receptors, Muscarinic, analysis, chemistry, immunology
Ämneskategorier Cell- och molekylärbiologi

Sammanfattning

We produced antibodies against a synthetic peptide corresponding to amino acids 168-192 of the second extracellular loop of the M2 human muscarinic receptor in rabbits. In immunoblot, affinity-purified antibodies specifically recognized a major band of rat ventricular muscarinic receptor protein with a molecular weight of about 80 KD. This recognition could be blocked by pre-incubation with peptide. Moreover, with both light (LM) and electron microscopic (EM) immunocytochemistry techniques, muscarinic receptors were detected on sarcolemma and T-tubules of rat cardiomyocytes. In addition, immunoreactions were localized in membranes of capillaries. Likewise, these reactivities were abolished by pre-incubation with peptide. These results suggest that the antibodies against the second extracellular loop of human M2 muscarinic receptor could specifically recognize rat ventricular muscarinic receptor protein and could be a powerful tool to study the fate of this receptor under different pathological or physiological conditions.

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