The Aryl Hydrocarbon Receptor Mediates UVB Radiation–Induced Skin Tanning Bettina Jux, Stephanie Kadow, Sandra Luecke, Agneta Rannug, Jean Krutmann, Charlotte.

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Volume 10, Issue 3, Pages (March 1999)

Presentation transcript:

The Aryl Hydrocarbon Receptor Mediates UVB Radiation–Induced Skin Tanning Bettina Jux, Stephanie Kadow, Sandra Luecke, Agneta Rannug, Jean Krutmann, Charlotte Esser Journal of Investigative Dermatology Volume 131, Issue 1, Pages 203-210 (January 2011) DOI: 10.1038/jid.2010.269 Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Normal mouse melanocytes express high levels of functional aryl hydrocarbon receptor (AhR). Mouse melanocytes were cultivated from epidermal skin as described. (a) Standard PCR and agarose gel electrophoresis of AhR, ARNT, and two housekeeping genes, HPRT and RPS6. S: molecular weight standard. One representative experiment out of three is shown. (b) Western blot: bars show densitometry of AhR expression standardized to GAPDH. 1: liver, 2: thymus, 3: wild-type (WT) melanocytes, 4: AhR-/- melanocytes. Primary cultivated melanocytes were exposed to 10nM 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (c), 1mM UVB-irradiated tryptophan (d), or 50–1000pM formylindolo(3,2-b)carbazole (FICZ) (e). Shown is n-fold induction of RNA for CYP1A1 after 24 hours, either in WT (black bars) or AhR-/- (white bars). The mean and SD values of three (TCDD, UVB-irr. L-trp) or two (FICZ dose) independent experiments are shown. ND, not detectable; UVB-irr. L-trp, UVB-irradiated L-tryptophan tryptophan. Journal of Investigative Dermatology 2011 131, 203-210DOI: (10.1038/jid.2010.269) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Tanning response of wild-type (WT) versus aryl hydrocarbon receptor (AhR)-deficient mice. (a) Photographs of epidermal sheets from tail skin of WT or AhR-/- mice, taken 2 weeks after a single irradiation of 180mJcm–2. The length of the biopsied tail pieces is approximately 1cm each. (b, c) Melanin content after 1 × 180mJcm–2 or 5 × 100mJcm–2 per week for 4 weeks. (d) Tyrosinase activity in tail epidermis. Three to six mice were used per group. Shown is a representative experiment. OD, optical density. Journal of Investigative Dermatology 2011 131, 203-210DOI: (10.1038/jid.2010.269) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Tanning response in K5Cre+ mice. (a) Aryl hydrocarbon receptor (AhR)flox/flox mice were bred with K5Cre mice. Keratinocyte-specific excision was confirmed with PCR. Epidermal cells (ECs) were stained with CD117, MHCII, and CD3 and separated by magnetic cell sorting. Lanes from left to right: (M) 100bp size standard (1) liver, (2) thymus, (3) enriched KC, (4) CD117/MHCII/CD3-enriched ECs, (5) unseparated epidermis. Wild-type (WT) band: 140bp; excised band: 180bp. (b) Melanin content in the ears of conditionally deficient mice (“K5Cre+”) in keratinocytes and in control mice having no AhR deletion (“K5Cre-”, exposed to 100mJcm–2 UVB five times per week for 4 weeks). (c) Tyrosinase activity in tail epidermis. At least four mice were used per group. Journal of Investigative Dermatology 2011 131, 203-210DOI: (10.1038/jid.2010.269) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 UVB-induced pigmentation. (a) Increase in dihydroxyphenylalanin (DOPA)+ cells in ear epidermis of wild-type (WT) versus aryl hydrocarbon receptor (AhR)-/- mice. WT and AhR-/- mice were irradiated with a single dose of 180mJcm–2, epidermis-prepared, and DOPA-stained. Scale bar=500μm. (a) Photographs of microscope fields in each group. (b) DOPA-positive ear melanocytes were counted using a light microscope, as described in Materials and Methods. Shown is mean+SD of 22 viewing fields. Five to six mice were used per group. Journal of Investigative Dermatology 2011 131, 203-210DOI: (10.1038/jid.2010.269) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Expression of stem cell factor (SCF) and c-kit in wild-type (WT) versus aryl hydrocarbon receptor (AhR)-/- melanocytes. (a) Primary melanocytes were cultured, harvested, and RNA-prepared for PCR of SCF and housekeeping gene Rps6; for SCF primers, see reference (Kawaguchi et al., 2001). The primers yield two products, soluble (sSCF) and membrane-bound SCF (mSCF). S: size marker. (b) Epidermal cells were prepared from WT and AhR-/- mice. Shown is a histogram of CD117 (c-kit) surface expression of cells negative for MHC-II, CD24, CD3, and CD16/32 to exclude Langerhans cells, keratinocytes, and T cells (left graph), and a bar diagram summarizing the results of six independent flow cytometry experiments (right graph). (c) Luciferase assay of c-kit promoter (two putative dioxin-responsive elements (DREs)) cloned into pGL3basic. Shown is the luminescence of HepG2 cells after transfection with the plasmid. As controls, results with transfections of the promoter of cyp1a1, which has five valid DREs, and the empty vector are shown. White bars: untransfected cells, black bars: transfected cells. For details on cloned fragments, see Supplementary Data S2 online. Journal of Investigative Dermatology 2011 131, 203-210DOI: (10.1038/jid.2010.269) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions