Differential expression of CRABP-II in fibroblasts derived from dermis and subcutaneous fat
Section snippets
Materials and methods
Tissue handling and cell culture. Dermal and subcutaneous fat tissues were obtained from healthy donors during abdominal dermolipectomy. A 0.3 mm split skin biopsy was taken using a dermatome (Aesculap AG & Co. KG, Tuttlingen, Germany). Pieces of subcutaneous fat were harvested using a scalpel. Large blood vessels, glandular tissue, and fascia were omitted. Part of the tissue was frozen in liquid nitrogen for RNA isolation. Dermis and fat were treated for cell isolation as described earlier [6].
Identification of differentially expressed proteins by 2D PAGE
The 2D-gel electrophoresis approach has been chosen to obtain information on the overall protein spectrum of cultured dermal and fat fibroblasts. The protein samples isolated from these cells were run on parallel 2D-gels and visualised using silver staining. Low inter-individual differences in profiles of proteins extracted from cultured cells derived from five different skin donors have been observed. Different spots were found to vary in intensity between images of dermal and fat fibroblast
Discussion
Wound healing in skin comprises a cascade of events during which the neo-dermis is formed and the wound surface is re-epithelialised. During re-epithelialisation keratinocytes migrate, proliferate, and differentiate on the formed neo-dermis. These processes are thought to be regulated amongst others by retinoids (e.g., retinoic acid and retinol). This specific group of vitamin A metabolites is reported to be vital for vision, (embryonic) growth, and reproduction [10]. They are essential in
References (29)
The cellular derivation and the life span of the myofibroblast
Pathol. Res. Pract.
(1996)Interactions of retinoid binding proteins and enzymes in retinoid metabolism
Biochim. Biophys. Acta
(1999)- et al.
Retinoids and state of differentiation modulate CRABP II gene expression in a skin equivalent
J. Invest. Dermatol.
(1993) Retinoic acid: its biosynthesis and metabolism
Prog. Nucleic Acid Res. Mol. Biol.
(1999)- et al.
Cellular retinol-binding protein-1 expression and modulation during in vivo and in vitro myofibroblastic differentiation of rat hepatic stellate cells and portal fibroblasts
Lab. Invest.
(2002) - et al.
Molecular cloning of two human cellular retinoic acid-binding proteins (CRABP). Retinoic acid-induced expression of CRABP-II but not CRABP-I in adult human skin in vivo and in skin fibroblasts in vitro
J. Biol. Chem.
(1991) - et al.
Differential regulation of retinoic acid receptors and binding proteins in human skin
J. Invest. Dermatol.
(1992) - et al.
Retinoid-binding proteins are phosphorylated in vitro by soluble Ca+2- and phosphatidylserine-dependent protein kinase from mouse brain
Biochem. Biophys. Res. Commun.
(1984) - et al.
Fibroblasts facilitate re-epithelialization in wounded human skin equivalents
Lab. Invest.
(2004) - et al.
Expression and myristoylation of NAP-22 using a baculovirus transfer vector system
Biochim. Biophys. Acta
(1994)
Natural N-terminal fragments of brain abundant myristoylated protein BASP1
Biochim. Biophys. Acta
Wound healing—aiming for perfect skin regeneration
Science
Extracellular matrix and keratinocyte migration
Clin. Exp. Dermatol.
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