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

Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3455-60. doi: 10.1073/pnas.93.8.3455.

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 for 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 beta-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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Calcitonin Gene-Related Peptide / pharmacology*
  • Chlorides / metabolism
  • Cloning, Molecular
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • DNA, Complementary / genetics
  • Female
  • Gene Expression
  • Guinea Pigs
  • Immunohistochemistry
  • In Vitro Techniques
  • Molecular Sequence Data
  • Oocytes / drug effects*
  • Oocytes / metabolism*
  • Organ of Corti / metabolism
  • RNA, Complementary / genetics
  • Receptors, Calcitonin Gene-Related Peptide / drug effects*
  • Receptors, Calcitonin Gene-Related Peptide / genetics*
  • Xenopus laevis

Substances

  • CRCP protein, human
  • Chlorides
  • DNA, Complementary
  • RNA, Complementary
  • Receptors, Calcitonin Gene-Related Peptide
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Calcitonin Gene-Related Peptide

Associated data

  • GENBANK/U50188