Deletion of μ- and κ-Opioid Receptors in Mice Changes Epidermal Hypertrophy, Density of Peripheral Nerve Endings, and Itch Behavior  Mei Bigliardi-Qi, Claire Gaveriaux-Ruff, Katrin Pfaltz, Pierre Bady, Tommy Baumann, Theo Rufli, Brigitte L. Kieffer, Paul L. Bigliardi  Journal of Investigative Dermatology  Volume 127, Issue 6, Pages 1479-1488 (June 2007) DOI: 10.1038/sj.jid.5700661 Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 PCR gels with the expected PCR product of 352bp for mouse MOR and 756bp for mouse KOR. Lanes a/e=negative control, lane b=mouse brain (MOR), lane c=skin of wild-type mouse (MOR), lane d=skin of MOR knockout mouse, lane f=mouse brain (KOR), lane g=skin of wild-type mouse (KOR), lane h=skin of KOR knockout mouse. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Reduced epidermal hypertrophy in MOR knockout mice compared to wild-type mice. Microscopic images of hematoxylin–eosin staining from the paraffin-embedded sections of mouse rostral back skin after 5 days treatment with AEW (acetone/diethylether/water). (a) Wild-type male, (b) MOR knockout male, (c) wild-type female, (d) MOR knockout female after induction of dry skin dermatitis, (e) untreated wild-type female, and (f) untreated MOR knockout female; bar=0.1mm. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Boxplot representation of epidermal hypertrophy. Pictures were taken from hematoxylin–eosin-stained paraffin sections from each animal and the epidermal thickness of each biopsy was measured at three different locations. The data of MOR and KOR knockout mice were compared to the wild-type mice after logarithmic transformation. Number of animals: wild-type untreated male (n=9)/female (n=10), wild-type treated male (n=14)/female (n=9), MOR knockout untreated male (n=7)/female (n=7), MOR knockout treated male (n=8)/female (n=8), KOR knockout untreated male (n=7)/female (n=7), KOR knockout treated male (n=6)/female (n=6). Panels compare the gender (a/b=male against c/d=female) with (b/d) treated and (a/c) untreated condition. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Boxplot representation of epidermal PGP 9.5 staining. Numbers of animals as in Figure 3. Panels compare the gender (a/b=male against c/d=female) with (b/d) treated and (a/c) untreated condition. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Representation of the relationship between epidermal thickness and epidermal PGP 9.5 staining. (a) Values grouped by type; (b) values grouped by treatment. The points correspond to the means in each individual and each point shows its standard deviation (both with logarithmic transformation). Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Boxplot representation of epidermal substance P staining. Wild-type (untreated: male (n=5)), female (n=6); treated: male (n=7), female (n=5) and MOR knockout (untreated: male (n=7), female (n=6); treated: male (n=8), female (n=8)) mice. Panels compare the gender (a/b=male against c/d=female) with (b/d) treated and (a/c) untreated condition. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 Representation of the relationship between epidermal staining of substance P and PGP 9.5. (a) Values grouped by type; (b) values grouped by treatment. The points correspond to the means by individual and each point is associated with standard deviations; substance P=square transformation, PGP 9.5=logarithmic transformation. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 8 Boxplot representation of CD4-positive T-lymphocytes in dermis. Formalin-fixed tissue from rostral back before and after induction of dry skin dermatitis in wild-type and MOR knockout mice (seven mice in each case). Panels compare the gender (a/b=male against c/d=female) with (b/d) treated and (a/c) untreated condition. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 9 Boxplot representation of counts of Giemsa-stain-positive mast cells in dermis. Formalin-fixed tissue from rostral back before and after induction of dry skin dermatitis in wild-type (untreated: male (n=9), female (n=11); treated: male (n=14), female (n=9) and MOR knockout (untreated: male (n=8), female (n=7); treated: male (n=8), female (n=8)) mice. Panels compare the gender (a/b=male against c/d=female) with (b/d) treated and (a/c) untreated condition. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 10 Boxplot representation of scratching behavior (log-transformation). The scratching behavior of the treated animals was observed by video taping over 1hour and the bouts of scratching were counted. The number of scratching bouts was divided by the number of minutes the mouse was awake during the time of the observation. Number of animals: wild-type mice (male (n=14), female (n=9)), MOR (male (n=8), female (n=8)) and KOR knockout mice (male (n=7), female (n=7)); (a) male, (b) female mice. Journal of Investigative Dermatology 2007 127, 1479-1488DOI: (10.1038/sj.jid.5700661) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions