Transcriptional Profiling Shows Altered Expression of Wnt Pathway– and Lipid Metabolism–Related Genes as Well as Melanogenesis-Related Genes in Melasma 

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Transcriptional Profiling Shows Altered Expression of Wnt Pathway– and Lipid Metabolism–Related Genes as Well as Melanogenesis-Related Genes in Melasma  Hee Young Kang, Itaru Suzuki, Dong Jun Lee, Jaehyun Ha, Pascale Reiniche, Jérôme Aubert, Sophie Deret, Didier Zugaj, Johannes J. Voegel, Jean-Paul Ortonne  Journal of Investigative Dermatology  Volume 131, Issue 8, Pages 1692-1700 (August 2011) DOI: 10.1038/jid.2011.109 Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Histochemical/immunohistochemical evaluations and messenger RNA levels of tyrosinase-related protein 1 (TYRP1). (a) Hematoxylin–eosin (HE) staining showed no major difference in the general morphology or cytology of skin samples. Fontana–Masson (FM) staining showed an obvious increase in epidermal melanin in lesional skin. In these subjects, we did not observe significant amounts of dermal melanin (bar=100μm). (b) Melanogenesis-associated factors (tyrosinase (TYR), TYRP1, dopachrome tautomerase (DCT), and silver (SILV)) and melanocyte-specific transcription factors (microphthalmia-associated transcription factor (MITF) and SOX10) were evaluated by immunohistochemistry. TYR is stained red and the other proteins are stained blue (see color figure online). The melanogenesis-associated factors were found to be upregulated in lesional skin (bar=100μm). (c) Numbers represent fold increases in lesional skin as compared with perilesional skin of same patients as determined by image analysis. Image analysis results corresponded well with visual assessments. Interestingly, numbers of melanocytes determined by MITF and SOX10 immunostaining were not significantly different. (d) Signals were observed in cells in the basal layer epidermis corresponding to the distribution of melanocytes. The mRNA levels of TYRP1 were elevated in lesional skin (bar=100μm). Journal of Investigative Dermatology 2011 131, 1692-1700DOI: (10.1038/jid.2011.109) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Hierarchical clustering. Hierarchical clustering analysis was performed on 131 probe sets found to be differentially expressed in lesional and perilesional skin. The results obtained revealed the existence of two subgroups, in which one subgroup of six patients (box) was more affected in terms of gene expressional changes than the other subgroup (four patients). Red and blue (see color figure online) represent up- and downregulated genes, respectively. Journal of Investigative Dermatology 2011 131, 1692-1700DOI: (10.1038/jid.2011.109) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Comparison of the barrier functions of lesional and perilesional skins. (a) Basal transepidermal water loss (TEWL) was not significantly different in lesional and perilesional skins. (b) TEWL values after barrier perturbation were significantly higher for lesional skin. (c) A significantly delayed barrier recovery rate was demonstrated by lesional skin. *P<0.05. Journal of Investigative Dermatology 2011 131, 1692-1700DOI: (10.1038/jid.2011.109) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Immunohistochemical staining of Wnt signaling-associated factors. WNT inhibitory factor 1 (WIF1), secreted frizzled-related protein 2 (SFRP2), and Wnt5a were immunohistochemically detected using specific antibodies. Protein levels of WIF1 were greater in melanocytes of lesional skin (a) than in those of perilesional skin (b). SFRP2 immunoreactivity around fibroblasts in the dermis seemed to be greater in lesional skin than in perilesional skin (c, d). Wnt5a showed greater expression in the basal layer and around fibroblasts in lesional skin (e) than in perilesional skin (f) (bar=200μm). Journal of Investigative Dermatology 2011 131, 1692-1700DOI: (10.1038/jid.2011.109) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions