Role of Peroxisome Proliferator-Activated Receptor α in Epidermal Development in Utero Matthias Schmuth, M.D., V.A., Kristina Schoonjans, Qian-Chun Yu,

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Role of Peroxisome Proliferator-Activated Receptor α in Epidermal Development in Utero Matthias Schmuth, M.D., V.A., Kristina Schoonjans, Qian-Chun Yu, Joachim W. Fluhr, Debra Crumrine, Jean-Pierre Hachem, Peggy Lau, Johan Auwerx, Peter M. Elias, Kenneth R. Feingold Journal of Investigative Dermatology Volume 119, Issue 6, Pages 1298-1303 (December 2002) DOI: 10.1046/j.1523-1747.2002.19605.x Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 On light microscopy SC development is delayed in PPAR-α–/– animals at day 18.5 of fetal skin development. Mouse skin sections (6 μm) were fixed in 4% formaldehyde and stained with hematoxylin and eosin. Photographs were taken with an Axiovision 2 Imaging Zeiss light microscope (Jena). Magnification bars: 32 μm. Journal of Investigative Dermatology 2002 119, 1298-1303DOI: (10.1046/j.1523-1747.2002.19605.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Electron microscopy reveals decreased number of SC layers in PPAR-α–/– animals. SC layers were assessed on ultrathin sections following OsO4 postfixation. Photographs were taken from fetal skin at gestational day 18.5. pd, periderm. Magnification bars: 0.5 μm. The number of SC layers was counted on toluidine-blue-stained semithin sections. Data are presented as mean ± SEM; n=3–5 (animals); **p < 0.05, ***p < 0.005. Journal of Investigative Dermatology 2002 119, 1298-1303DOI: (10.1046/j.1523-1747.2002.19605.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Processing of extracellular, lamellar bilayers is delayed, but the lamellar body secretory system is normal in PPAR-α–/– animals. (A), (B) There is a delay in processing at the first extracellular space outside the stratum granulosum–SC interface in PPAR-α–/– mice at day 18.5. Abnormal processing of secreted lamellar body contents into bilayer structures is indicated by arrows. (C) Stretches of intact, normal-appearing bilayers are present (arrowheads) in the SC of PPAR-α+/+ mice. RuO4 postfixation. SG, stratum granulosum. Magnification bars: 0.5 μm. Journal of Investigative Dermatology 2002 119, 1298-1303DOI: (10.1046/j.1523-1747.2002.19605.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Cornified envelope protein expression is modestly reduced in PPAR-α–/– animals. Loricrin expression was assessed by immunohistochemistry on skin of day 18.5 fetuses (left panel) and at birth (right panel) as described in Methods. The micrographs shown were taken from ventral skin of the fetuses comparing PPAR-α–/– mice to wildtype mice (+/+). Magnification bars: 16 μm. Journal of Investigative Dermatology 2002 119, 1298-1303DOI: (10.1046/j.1523-1747.2002.19605.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 β-Glucocerebrosidase activity is diminished in PPAR-α–/– animals. Skin sections from day 18.5 pups were overlayered with β-glucocerebrosidase substrate solution (10 mM Resorufin β-D-glucopyranoside), incubated for 16 h at 4°C, and visualized with confocal microscopy. The left panel shows PPAR-α–/– skin and the right panel skin from a wildtype mouse (+/+). Magnification bars: 90 μm. Journal of Investigative Dermatology 2002 119, 1298-1303DOI: (10.1046/j.1523-1747.2002.19605.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Putative actions of extracutaneous and endogenous PPAR-α activators during epidermal development. Journal of Investigative Dermatology 2002 119, 1298-1303DOI: (10.1046/j.1523-1747.2002.19605.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions