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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
Author(s) -
Choksi Tejal,
Hay Debbie L,
Legon Stephen,
Poyner David R,
Hagner Stefanie,
Bloom Stephen R,
Smith David M
Publication year - 2002
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1038/sj.bjp.0704761
Subject(s) - adrenomedullin , calcitonin gene related peptide , receptor , medicine , endocrinology , calcitonin , calcitonin receptor , chemistry , cell culture , biology , microbiology and biotechnology , neuropeptide , genetics
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 (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. 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