Abstract
The expression of the messenger RNAs coding for glucagon-like peptide-I (GLP-I) receptor, VIP receptor, and pituitary adenylate cyclase-activating polypeptide (PACAP) receptor as well as the expression of the receptor proteins were demonstrated in the rat medullary carcinoma of thyroid cell line 6/23 by the following experiments: 1) RNA extraction, reverse transcriptase, and polymerase chain reaction with specific primers; 2) binding of the radiolabeled ligands [125I]GLP-I-(7-36)-NH2, [125I]PACAP-(1-27), and [125I]VIP and inhibition by, respectively, unlabeled GLP-I-(7-36)-NH2, PACAP-(1-27), and VIP; and 3) study of adenylate cyclase activation by the peptides and selective inhibition of the VIP/PACAP response by the antagonist [D-Phe2]VIP. Besides the highly selective GLP-I receptor, PACAP receptors of types I and II were present on the cell line and coupled to adenylate cyclase. PACAP stimulated the adenylate cyclase through type I and II receptors, whereas VIP interacted with type II receptors only. Messenger RNA analysis indicated that at least three splice variants of the PACAP type I receptor may be expressed in 6/23 cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenylyl Cyclases / analysis
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Animals
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Base Sequence
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Carcinoma, Medullary / chemistry*
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Carcinoma, Medullary / genetics
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Carcinoma, Medullary / pathology*
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DNA, Neoplasm / analysis
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DNA, Neoplasm / genetics
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Gene Expression Regulation, Neoplastic
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Glucagon-Like Peptide-1 Receptor
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Iodine Radioisotopes
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Molecular Sequence Data
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Polymerase Chain Reaction / methods
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RNA, Messenger / analysis
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RNA, Messenger / genetics
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Rats
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Receptors, Cell Surface / analysis*
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Receptors, Cell Surface / genetics
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Receptors, Cell Surface / physiology
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Receptors, Glucagon*
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Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide
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Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
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Receptors, Pituitary Hormone / analysis*
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Receptors, Pituitary Hormone / genetics
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Receptors, Pituitary Hormone / physiology
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Receptors, Vasoactive Intestinal Peptide / analysis
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Receptors, Vasoactive Intestinal Peptide / genetics
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Receptors, Vasoactive Intestinal Peptide / physiology
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Thyroid Neoplasms / chemistry*
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Thyroid Neoplasms / genetics
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Thyroid Neoplasms / pathology*
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Tumor Cells, Cultured
Substances
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DNA, Neoplasm
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Glp1r protein, rat
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Glucagon-Like Peptide-1 Receptor
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Iodine Radioisotopes
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RNA, Messenger
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Receptors, Cell Surface
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Receptors, Glucagon
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Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide
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Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
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Receptors, Pituitary Hormone
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Receptors, Vasoactive Intestinal Peptide
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Adenylyl Cyclases