Persistence of Both Peripheral and Non-Peripheral Corneodesmosomes in the Upper Stratum Corneum of Winter Xerosis Skin Versus only Peripheral in Normal.

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Persistence of Both Peripheral and Non-Peripheral Corneodesmosomes in the Upper Stratum Corneum of Winter Xerosis Skin Versus only Peripheral in Normal Skin Dominique Bernard, Anne-Marie Minondo, Christiane Camus, Françoise Fiat, Pierre Corcuff, Rainer Schmidt Journal of Investigative Dermatology Volume 116, Issue 1, Pages 23-30 (January 2001) DOI: 10.1046/j.1523-1747.2001.00208.x Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Various amounts of corneodesmosomal proteins are observed in upper SC extracts. Equal amounts of total proteins corresponding to three serial strippings performed on one individual with normal skin (NS, lanes 1–3) and on one individual with xerotic skin (XS, lanes 4–6), and, as a control, proteins of a Tris-urea buffer extract of breast epidermis (lane 7) were analyzed. The proteins were separated by SDS-PAGE, stained with Coomassie blue (a), or transferred to nitrocellulose membranes and immunoblotted with (b) DG3.10, a MoAb directed to Dsg1 and Dsg2, (c) G36 -19, a MoAb directed to Cdsn, and (d) PG5.1, a MoAb directed to plakoglobin. The position of molecular mass standards (kDa) is indicated on the left. The arrows show entire 160 kDa Dsg1 (b), the 52–56 kDa Cdsn (c), and entire 83 kDa plakoglobin (Plak, d). Journal of Investigative Dermatology 2001 116, 23-30DOI: (10.1046/j.1523-1747.2001.00208.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 The amounts of Dsg1, plakoglobin, and Cdsn are increased in the SC extracts of individuals with winter xerosis skin. Equal amounts of total proteins corresponding to the first stripping performed on the 56 individuals were analyzed by western blotting with DG3.10, a MoAb directed to Dsg1, with a mix of PG5.1 and an anti-γ-catenin antibody, two MoAb directed to plakoglobin, and with a mix of G36-19 and F28-27, two MoAb directed to Cdsn. The immunoreactive bands were then quantified by densitometry. The values obtained for the 26 individuals with normal skin (NS) and the 30 individuals with xerotic skin (XS) are represented by squares, on an arbitrary scale. The horizontal lines correspond to the calculated medians. Note the significantly increased amounts of Dsg1, plakoglobin, and Cdsn in xerotic skin compared with normal skin (*p =0.05; **p <0.02). Journal of Investigative Dermatology 2001 116, 23-30DOI: (10.1046/j.1523-1747.2001.00208.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Freeze-fracture replicas show an increased number of corneodesmosomes on the surface of corneocytes from the outer SC of winter xerosis skin compared with those of normal skin. The morphology of intercellular spaces within the inner SC (a, c) and outer SC (b, d) from normal (a, b) and xerotic (c, d) skin strippings was analyzed on freeze-fracture electron micrographs. Corneodesmosomes appear as particle aggregates (asterisks and stars). Arrowheads indicate the direction of shadowing. Scale bar: 250 nm. Journal of Investigative Dermatology 2001 116, 23-30DOI: (10.1046/j.1523-1747.2001.00208.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Freeze-fracture-labeled preparations of inner SC show Cdsn within the corneodesmosomes. Freeze-fractured samples of SC were immunogold labeled in the presence or absence of G36-19 MoAb. On platinum/carbon replicas of the fracture-labeled samples, G36-19 labeling is specifically detected on the particle aggregates morphologically identifiable as corneodesmosomes (a), whereas no labeling is observed in the absence of primary antibody (b). On electron micrographs of thin cross-sections of the same fracture-labeled samples, G36-19 labeling is specifically observed at the external surface of the samples in the core of corneodesmosomes (c), whereas no labeling is observed in the absence of primary antibody (d). The inset in (c) shows a higher magnification of a labeled corneodesmosome. Scale bars are shown on the micrographs. Journal of Investigative Dermatology 2001 116, 23-30DOI: (10.1046/j.1523-1747.2001.00208.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Quantification of corneodesmosome density at the surface of corneocytes confirms the reduced degradation of the structures in the outer SC of winter xerosis skin. The corneodesmosome density (area occupied by corneodesmosomes divided by total area of the micrograph) was measured on electron micrographs of the inner SC (two micrographs per individual) and the outer SC (four micrographs per individual) of normal (NS, n = 2) and winter xerosis (XS, n = 3) skin samples. Histograms representing the mean values show similar densities of corneodesmosomes in the outer SC of xerotic and normal skin. The density of corneodesmosomes in the upper SC of xerotic skin, however, was significantly increased compared with normal skin (p <0.001). Bars, standard deviations. Journal of Investigative Dermatology 2001 116, 23-30DOI: (10.1046/j.1523-1747.2001.00208.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Conventional transmission electron microscopy confirms the persistence of corneodesmosomes in the outer SC of winter xerosis skin. Varnish-strippings of normal (a, c) and winter xerosis (b, d) skin were analyzed by conventional transmission electron microscopy. Note that, when combined, the micrographs of normal SC correspond to the whole sample. Only part of the total height of xerosis SC, which is much thicker, is shown, however. In the outer SC of normal skin (a), corneocytes are loose and corneodesmosomes (arrows) are scarcely observed. In the outer SC of xerotic skin (b), corneocytes are more cohesive and corneodesmosomes are numerous. In the inner SC of both normal (c) and xerotic (d) skin, corneodesmosomes are numerous. Arrowheads indicate the outer surface of the samples. Scale bar: 0.5 μm. Journal of Investigative Dermatology 2001 116, 23-30DOI: (10.1046/j.1523-1747.2001.00208.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions