Identification of a protein that confers calcitonin gene-related peptide responsiveness to oocytes by using a cystic fibrosis transmembrane conductance regulator assay

Anne E. Luebke, Gerhard Dahl, Bernard A. Roos, Ian M. Dickerson

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

An expression-cloning strategy was used to isolate a cDNA that encodes a protein that confers calcitonin gene-related peptide (CGRP) responsiveness to Xenopus laevis oocytes. A guinea pig organ of Corti (the mammalian hearing organ) cDNA library was screened by using an assay based on the cystic fibrosis transmembrane conductance regulator (CFTR). The CFTR is a chloride channel that is activated upon phosphorylation; this channel activity was used as a sensor far CGRP-induced activation of intracellular kinases. A cDNA library from guinea pig organ of Corti was screened by using this oocyte- CFTR assay. A cDNA was identified that contained an open reading frame coding for a small hydrophilic protein that is presumed to be either a CGRP receptor or a component of a CGRP receptor complex. This CGRP receptor component protein confers CGRP-specific activation to the CFTR assay, as no activation was detected upon application of calcitonin, amylin, neuropeptide Y, vasoactive intestinal peptide, or β-endorphin. In situ hybridization demonstrated that the CGRP receptor component protein is expressed in outer hair cells of the organ of Corti and is colocalized with CGRP-containing efferent nerve terminals.

Original languageEnglish
Pages (from-to)3455-3460
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number8
DOIs
StatePublished - Apr 16 1996

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Calcitonin Gene-Related Peptide Receptors
Cystic Fibrosis Transmembrane Conductance Regulator
Calcitonin Gene-Related Peptide
Organ of Corti
Oocytes
Gene Library
Outer Auditory Hair Cells
Guinea Pigs
Proteins
Complementary DNA
Islet Amyloid Polypeptide
Endorphins
Chloride Channels
Neuropeptide Y
Vasoactive Intestinal Peptide
Xenopus laevis
Calcitonin
Open Reading Frames
Hearing
In Situ Hybridization

Keywords

  • cochlea
  • expression cloning
  • guinea pig

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

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abstract = "An expression-cloning strategy was used to isolate a cDNA that encodes a protein that confers calcitonin gene-related peptide (CGRP) responsiveness to Xenopus laevis oocytes. A guinea pig organ of Corti (the mammalian hearing organ) cDNA library was screened by using an assay based on the cystic fibrosis transmembrane conductance regulator (CFTR). The CFTR is a chloride channel that is activated upon phosphorylation; this channel activity was used as a sensor far CGRP-induced activation of intracellular kinases. A cDNA library from guinea pig organ of Corti was screened by using this oocyte- CFTR assay. A cDNA was identified that contained an open reading frame coding for a small hydrophilic protein that is presumed to be either a CGRP receptor or a component of a CGRP receptor complex. This CGRP receptor component protein confers CGRP-specific activation to the CFTR assay, as no activation was detected upon application of calcitonin, amylin, neuropeptide Y, vasoactive intestinal peptide, or β-endorphin. In situ hybridization demonstrated that the CGRP receptor component protein is expressed in outer hair cells of the organ of Corti and is colocalized with CGRP-containing efferent nerve terminals.",
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AU - Dahl, Gerhard

AU - Roos, Bernard A.

AU - Dickerson, Ian M.

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