Comparison of the expression of calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMPs) with CGRP and adrenomedullin binding in cell lines

Tejal Choksi, Debbie L. Hay, Stephen Legon, David R. Poyner, Stefanie Hagner, Stephen R. Bloom, David M. Smith

Research output: Contribution to journalArticle

Abstract

1. The calcitonin receptor-like receptor (CRLR) and specific receptor activity modifying proteins (RAMPs) together form receptors for calcitonin gene-related peptide (CGRP) and/or adrenomedullin in transfected cells. 2. There is less evidence that innate CGRP and adrenomedullin receptors are formed by CRLR/RAMP combinations. We therefore examined whether CGRP and/or adrenomedullin binding correlated with CRLR and RAMP mRNA expression in human and rat cell lines known to express these receptors. Specific human or rat CRLR antibodies were used to examine the presence of CRLR in these cells. 3. We confirmed CGRP subtype 1 receptor (CGRP(1)) pharmacology in SK-N-MC neuroblastoma cells. L6 myoblast cells expressed both CGRP(1) and adrenomedullin receptors whereas Rat-2 fibroblasts expressed only adrenomedullin receptors. In contrast we could not confirm CGRP(2) receptor pharmacology for Col-29 colonic epithelial cells, which, instead were CGRP(1)-like in this study. 4. L6, SK-N-MC and Col-29 cells expressed mRNA for RAMP1 and RAMP2 but Rat-2 fibroblasts had only RAMP2. No cell line had detectable RAMP3 mRNA. 5. SK-N-MC, Col-29 and Rat-2 fibroblast cells expressed CRLR mRNA. By contrast, CRLR mRNA was undetectable by Northern analysis in one source of L6 cells. Conversely, a different source of L6 cells had mRNA for CRLR. All of the cell lines expressed CRLR protein. Thus circumstances where CRLR mRNA is apparently absent by Northern analysis do not exclude the presence of this receptor. 6. These data strongly support CRLR, together with appropriate RAMPs as binding sites for CGRP and adrenomedullin in cultured cells.
Original languageEnglish
Pages (from-to)784-792
Number of pages9
JournalBritish Journal of Pharmacology
Volume136
Issue number5
DOIs
Publication statusPublished - 27 Jul 2002

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Receptor Activity-Modifying Proteins
Calcitonin Receptor-Like Protein
Adrenomedullin
Calcitonin Gene-Related Peptide
Cell Line
Calcitonin Gene-Related Peptide Receptors
Adrenomedullin Receptors
Messenger RNA
Fibroblasts
Pharmacology
Myoblasts
Neuroblastoma
Protein Binding

Keywords

  • animals
  • calcitonin
  • gene-related peptide
  • carrier
  • proteins
  • cell
  • line
  • DNA-binding proteins
  • humans
  • peptide
  • protein binding rats receptors
  • calcitonin receptors
  • peptide transcription factors
  • CGRP
  • adrenomedullin
  • CRLR
  • RAMP
  • SK-N-MC neuroblastoma cells
  • Col-29 cells
  • L6 myoblasts
  • Rat-2 fibroblasts

Cite this

Choksi, Tejal ; Hay, Debbie L. ; Legon, Stephen ; Poyner, David R. ; Hagner, Stefanie ; Bloom, Stephen R. ; Smith, David M. / Comparison of the expression of calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMPs) with CGRP and adrenomedullin binding in cell lines. In: British Journal of Pharmacology. 2002 ; Vol. 136, No. 5. pp. 784-792.
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Comparison of the expression of calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMPs) with CGRP and adrenomedullin binding in cell lines. / Choksi, Tejal; Hay, Debbie L.; Legon, Stephen; Poyner, David R.; Hagner, Stefanie; Bloom, Stephen R.; Smith, David M.

In: British Journal of Pharmacology, Vol. 136, No. 5, 27.07.2002, p. 784-792.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Comparison of the expression of calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMPs) with CGRP and adrenomedullin binding in cell lines

AU - Choksi, Tejal

AU - Hay, Debbie L.

AU - Legon, Stephen

AU - Poyner, David R.

AU - Hagner, Stefanie

AU - Bloom, Stephen R.

AU - Smith, David M.

PY - 2002/7/27

Y1 - 2002/7/27

N2 - 1. The calcitonin receptor-like receptor (CRLR) and specific receptor activity modifying proteins (RAMPs) together form receptors for calcitonin gene-related peptide (CGRP) and/or adrenomedullin in transfected cells. 2. There is less evidence that innate CGRP and adrenomedullin receptors are formed by CRLR/RAMP combinations. We therefore examined whether CGRP and/or adrenomedullin binding correlated with CRLR and RAMP mRNA expression in human and rat cell lines known to express these receptors. Specific human or rat CRLR antibodies were used to examine the presence of CRLR in these cells. 3. We confirmed CGRP subtype 1 receptor (CGRP(1)) pharmacology in SK-N-MC neuroblastoma cells. L6 myoblast cells expressed both CGRP(1) and adrenomedullin receptors whereas Rat-2 fibroblasts expressed only adrenomedullin receptors. In contrast we could not confirm CGRP(2) receptor pharmacology for Col-29 colonic epithelial cells, which, instead were CGRP(1)-like in this study. 4. L6, SK-N-MC and Col-29 cells expressed mRNA for RAMP1 and RAMP2 but Rat-2 fibroblasts had only RAMP2. No cell line had detectable RAMP3 mRNA. 5. SK-N-MC, Col-29 and Rat-2 fibroblast cells expressed CRLR mRNA. By contrast, CRLR mRNA was undetectable by Northern analysis in one source of L6 cells. Conversely, a different source of L6 cells had mRNA for CRLR. All of the cell lines expressed CRLR protein. Thus circumstances where CRLR mRNA is apparently absent by Northern analysis do not exclude the presence of this receptor. 6. These data strongly support CRLR, together with appropriate RAMPs as binding sites for CGRP and adrenomedullin in cultured cells.

AB - 1. The calcitonin receptor-like receptor (CRLR) and specific receptor activity modifying proteins (RAMPs) together form receptors for calcitonin gene-related peptide (CGRP) and/or adrenomedullin in transfected cells. 2. There is less evidence that innate CGRP and adrenomedullin receptors are formed by CRLR/RAMP combinations. We therefore examined whether CGRP and/or adrenomedullin binding correlated with CRLR and RAMP mRNA expression in human and rat cell lines known to express these receptors. Specific human or rat CRLR antibodies were used to examine the presence of CRLR in these cells. 3. We confirmed CGRP subtype 1 receptor (CGRP(1)) pharmacology in SK-N-MC neuroblastoma cells. L6 myoblast cells expressed both CGRP(1) and adrenomedullin receptors whereas Rat-2 fibroblasts expressed only adrenomedullin receptors. In contrast we could not confirm CGRP(2) receptor pharmacology for Col-29 colonic epithelial cells, which, instead were CGRP(1)-like in this study. 4. L6, SK-N-MC and Col-29 cells expressed mRNA for RAMP1 and RAMP2 but Rat-2 fibroblasts had only RAMP2. No cell line had detectable RAMP3 mRNA. 5. SK-N-MC, Col-29 and Rat-2 fibroblast cells expressed CRLR mRNA. By contrast, CRLR mRNA was undetectable by Northern analysis in one source of L6 cells. Conversely, a different source of L6 cells had mRNA for CRLR. All of the cell lines expressed CRLR protein. Thus circumstances where CRLR mRNA is apparently absent by Northern analysis do not exclude the presence of this receptor. 6. These data strongly support CRLR, together with appropriate RAMPs as binding sites for CGRP and adrenomedullin in cultured cells.

KW - animals

KW - calcitonin

KW - gene-related peptide

KW - carrier

KW - proteins

KW - cell

KW - line

KW - DNA-binding proteins

KW - humans

KW - peptide

KW - protein binding rats receptors

KW - calcitonin receptors

KW - peptide transcription factors

KW - CGRP

KW - adrenomedullin

KW - CRLR

KW - RAMP

KW - SK-N-MC neuroblastoma cells

KW - Col-29 cells

KW - L6 myoblasts

KW - Rat-2 fibroblasts

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U2 - 10.1038/sj.bjp.0704761

DO - 10.1038/sj.bjp.0704761

M3 - Article

VL - 136

SP - 784

EP - 792

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 5

ER -