Document Detail


Quantitative analysis of MC1R gene expression in human skin cell cultures.
MedLine Citation:
PMID:  16420249     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To address the issue of melanocortin-1 receptor (MC1R) expression in non-melanocytic cells, we have quantitatively evaluated the relative expression levels of both MC1R mRNA and protein in a subset of different cell types. Using semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) at high cycle numbers, we detected MC1R mRNA in all cell types examined, including human embryonic kidney-293 (HEK 293) cells, a cell type widely used as a negative control in melanocortin expression studies. Quantitative real-time PCR revealed the highest levels of MC1R transcripts were in melanocytic cells, whereas the keratinocyte and fibroblast cell cultures examined had only a low level of expression, similar to that of HEK 293 cells. Antibody mediated detection of MC1R protein in membrane extracts demonstrated exogenous receptor in MC1R transfected cell lines, as well as endogenous MC1R in melanoma cells. However, radioligand binding procedures were required to detect MC1R protein of normal human melanocytes and no surface expression of MC1R was detected in any of the non-melanocytic cells examined. This was consistent with their low level of mRNA, and suggests that, if present, the levels of surface receptor are significantly lower than that in melanocytes. The capacity of such limited levels of MC1R protein to influence non-melanocytic skin cell biology would likely be severely compromised. Indeed, the MC1R agonist [NIe(4), D-Phe(7)] alpha-melanocyte stimulating hormone (NDP-MSH) was unable to elevate intracellular cyclic adenosine monophosphate (cAMP) levels in the keratinocyte and fibroblast cells examined, whereas a robust increase was elicited in melanocytes. Although there are a variety of cell types with detectable MC1R mRNA, the expression of physiologically significant levels of the receptor may be more restricted than the current literature indicates, and within epidermal tissue may be limited to the melanocyte.
Authors:
Donald W Roberts; Richard A Newton; Kimberley A Beaumont; J Helen Leonard; Richard A Sturm
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society     Volume:  19     ISSN:  0893-5785     ISO Abbreviation:  Pigment Cell Res.     Publication Date:  2006 Feb 
Date Detail:
Created Date:  2006-01-19     Completed Date:  2006-04-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8800247     Medline TA:  Pigment Cell Res     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  76-89     Citation Subset:  IM    
Affiliation:
Melanogenix Group, Institute for Molecular Bioscience, University of Queensland, Brisbane, Qld 4072, Australia.
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MeSH Terms
Descriptor/Qualifier:
Cells, Cultured
Cyclic AMP / metabolism
Fibroblasts / cytology,  metabolism
Gene Expression Profiling
Gene Expression Regulation*
Humans
Keratinocytes / cytology,  metabolism
Melanocyte-Stimulating Hormones / metabolism
Melanocytes / cytology,  metabolism
Polymerase Chain Reaction / methods
RNA, Messenger / metabolism
Radioligand Assay
Receptor, Melanocortin, Type 1 / genetics*,  metabolism*
Skin / cytology*
Chemical
Reg. No./Substance:
0/RNA, Messenger; 0/Receptor, Melanocortin, Type 1; 60-92-4/Cyclic AMP; 9002-79-3/Melanocyte-Stimulating Hormones

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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