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

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. There is less evidence that innate CGRP and adrenomedullin receptors are formed by CRL...

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Veröffentlicht in:British journal of pharmacology 2002-07, Vol.136 (5), p.784-792
Hauptverfasser: Choksi, Tejal, Hay, Debbie L, Legon, Stephen, Poyner, David R, Hagner, Stefanie, Bloom, Stephen R, Smith, David M
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container_issue 5
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container_title British journal of pharmacology
container_volume 136
creator Choksi, Tejal
Hay, Debbie L
Legon, Stephen
Poyner, David R
Hagner, Stefanie
Bloom, Stephen R
Smith, David M
description 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. 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. We confirmed CGRP subtype 1 receptor (CGRP1) pharmacology in SK‐N‐MC neuroblastoma cells. L6 myoblast cells expressed both CGRP1 and adrenomedullin receptors whereas Rat‐2 fibroblasts expressed only adrenomedullin receptors. In contrast we could not confirm CGRP2 receptor pharmacology for Col‐29 colonic epithelial cells, which, instead were CGRP1‐like in this study. 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. 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. These data strongly support CRLR, together with appropriate RAMPs as binding sites for CGRP and adrenomedullin in cultured cells. British Journal of Pharmacology (2002) 136, 784–792; doi:10.1038/sj.bjp.0704761
doi_str_mv 10.1038/sj.bjp.0704761
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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. These data strongly support CRLR, together with appropriate RAMPs as binding sites for CGRP and adrenomedullin in cultured cells. 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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. We confirmed CGRP subtype 1 receptor (CGRP1) pharmacology in SK‐N‐MC neuroblastoma cells. L6 myoblast cells expressed both CGRP1 and adrenomedullin receptors whereas Rat‐2 fibroblasts expressed only adrenomedullin receptors. In contrast we could not confirm CGRP2 receptor pharmacology for Col‐29 colonic epithelial cells, which, instead were CGRP1‐like in this study. 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. 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. These data strongly support CRLR, together with appropriate RAMPs as binding sites for CGRP and adrenomedullin in cultured cells. British Journal of Pharmacology (2002) 136, 784–792; doi:10.1038/sj.bjp.0704761</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>12086988</pmid><doi>10.1038/sj.bjp.0704761</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record>
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subjects Adrenomedullin
Animals
Calcitonin Gene-Related Peptide - metabolism
Calcitonin Receptor-Like Protein
Carrier Proteins - biosynthesis
Cell Line
CGRP
Col‐29 cells
CRLR
DNA-Binding Proteins - biosynthesis
Humans
L6 myoblasts
Peptides - metabolism
Protein Binding - physiology
RAMP
Rats
Rat‐2 fibroblasts
Receptors, Adrenomedullin
Receptors, Calcitonin - biosynthesis
Receptors, Peptide - metabolism
SK‐N‐MC neuroblastoma cells
Transcription Factors
title Comparison of the expression of calcitonin receptor‐like receptor (CRLR) and receptor activity modifying proteins (RAMPs) with CGRP and adrenomedullin binding in cell lines
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