PMID- 16420249
OWN - NLM
STAT- MEDLINE
DCOM- 20060414
LR  - 20161021
IS  - 0893-5785 (Print)
IS  - 0893-5785 (Linking)
VI  - 19
IP  - 1
DP  - 2006 Feb
TI  - Quantitative analysis of MC1R gene expression in human skin cell cultures.
PG  - 76-89
AB  - 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.
FAU - Roberts, Donald W
AU  - Roberts DW
AD  - Melanogenix Group, Institute for Molecular Bioscience, University of Queensland, 
      Brisbane, Qld 4072, Australia.
FAU - Newton, Richard A
AU  - Newton RA
FAU - Beaumont, Kimberley A
AU  - Beaumont KA
FAU - Helen Leonard, J
AU  - Helen Leonard J
FAU - Sturm, Richard A
AU  - Sturm RA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Denmark
TA  - Pigment Cell Res
JT  - Pigment cell research
JID - 8800247
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptor, Melanocortin, Type 1)
RN  - 9002-79-3 (Melanocyte-Stimulating Hormones)
RN  - E0399OZS9N (Cyclic AMP)
SB  - IM
MH  - Cells, Cultured
MH  - Cyclic AMP/metabolism
MH  - Fibroblasts/cytology/metabolism
MH  - Gene Expression Profiling
MH  - *Gene Expression Regulation
MH  - Humans
MH  - Keratinocytes/cytology/metabolism
MH  - Melanocyte-Stimulating Hormones/metabolism
MH  - Melanocytes/cytology/metabolism
MH  - Polymerase Chain Reaction/methods
MH  - RNA, Messenger/metabolism
MH  - Radioligand Assay
MH  - Receptor, Melanocortin, Type 1/*genetics/*metabolism
MH  - Skin/*cytology
EDAT- 2006/01/20 09:00
MHDA- 2006/04/15 09:00
CRDT- 2006/01/20 09:00
PHST- 2006/01/20 09:00 [pubmed]
PHST- 2006/04/15 09:00 [medline]
PHST- 2006/01/20 09:00 [entrez]
AID - PCR286 [pii]
AID - 10.1111/j.1600-0749.2005.00286.x [doi]
PST - ppublish
SO  - Pigment Cell Res. 2006 Feb;19(1):76-89. doi: 10.1111/j.1600-0749.2005.00286.x.