Differential expression of CRABP-II in fibroblasts derived from dermis and subcutaneous fat.
We have shown previously that fibroblasts derived from fat or dermal tissue differ in their functional properties, such as proliferation rate and contractile properties. To study these differences further, two-dimensional electrophoresis (2D PAGE) was performed on proteins isolated from cultured subcutaneous fat and dermal fibroblasts. The 2D gels were screened for proteins that were differentially expressed in all donors (n = 5). Five protein spots were subjected to further analysis by mass spectrometry. Two proteins could be identified: brain acid soluble protein 1 (BASP1) and cellular retinoic acid binding protein-II (CRABP-II). CRABP-II is of interest in terms of re-epithelialisation and was clearly expressed in dermal fibroblasts but not in fat fibroblasts. Real time PCR was performed to confirm the 2D data on CRABP-II. The CRABP-II mRNA level was significantly increased in dermal tissue and cultured dermal fibroblasts compared to fat tissue and cultured fat-derived fibroblasts, respectively. The mode of action of CRABP-II in skin is to mediate retinoic acid activity. Retinoic acid is known to inhibit migration and to stimulate differentiation of keratinocytes. The expression of CRABP-II by dermal fibroblasts implicates a role for these fibroblasts in wound re-epithelialisation, in contrast to subcutaneous fat-derived fibroblasts.
Van den Bogaerdt
Antoon J
AJ
Research Department of the Dutch Burns Foundation, 1940 EA Beverwijk, The Netherlands. AvdBogaerdt@dbf.nl
El Ghalbzouri
Abdoelwaheb
A
Hensbergen
Paul J
PJ
Reijnen
Linda
L
Verkerk
Michelle
M
Kroon-Smits
Miriam
M
Middelkoop
Esther
E
Ulrich
Magda M W
MM
D000273:Adipose Tissue / Q000378:metabolism*; D001483:Base Sequence; D002454:Cell Differentiation; D002465:Cell Movement; D017931:DNA Primers / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D005347:Fibroblasts / Q000378:metabolism*; D005786:Gene Expression Regulation; D006863:Hydrogen-Ion Concentration; D015603:Keratinocytes / Q000378:metabolism; D013058:Mass Spectrometry; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D018168:Receptors, Retinoic Acid / Q000096:biosynthesis* / Q000737:chemistry*; D012097:Repressor Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000378:metabolism*; D014018:Tissue Distribution; D014212:Tretinoin / Q000378:metabolism
C104089:BASP1 protein, human; D017931:DNA Primers; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D010455:Peptides; D012333:RNA, Messenger; D018168:Receptors, Retinoic Acid; D012097:Repressor Proteins; C082928:retinoic acid binding protein II, cellular; D014212:Tretinoin; D012313:RNA