Epidermal keratinocytes form a functional skin barrier in the absence of Atg7 dependent autophagy

• Published in: Journal of dermatological science 2013-07, Vol.71 (1), p.67-75
• Main Authors: Rossiter, Heidemarie, König, Ulrich, Barresi, Caterina, Buchberger, Maria, Ghannadan, Minoo, Zhang, Cheng-Feng, Mlitz, Veronika, Gmeiner, Ramona, Sukseree, Supawadee, Födinger, Dagmar, Eckhart, Leopold, Tschachler, Erwin
• Format: Article
• Language: English
• Subjects: Animals; Autophagy; Autophagy-Related Protein 7; Cell Differentiation; Cells, Cultured; Dermatology; Diffusion; Epidermis; Epidermis - metabolism; Epidermis - ultrastructure; GFP–LC3; Green Fluorescent Proteins; Keratin-14 - genetics; Keratinocytes; Keratinocytes - metabolism; Keratinocytes - ultrastructure; Mice; Mice, Inbred CBA; Mice, Knockout; Mice, Transgenic; Microtubule-Associated Proteins - deficiency; Microtubule-Associated Proteins - genetics; Microtubule-Associated Proteins - metabolism; Permeability; Promoter Regions, Genetic; Skin Absorption; Water Loss, Insensible
• ISSN: 0923-1811; 1873-569X
• DOI: 10.1016/j.jdermsci.2013.04.015

Abstract Background Cornification of keratinocytes involves the degradation of intracellular constituents which has led to the hypothesis that autophagy plays a role in this process. Mice, in which essential autophagy-related genes such as Atg7 are deleted systemically, die after birth and have not been characterized for potential epidermal defects. Objective This study tested whether autophagy is essential for epidermal barrier formation and function. Methods Atg7 was inactivated in epidermal keratinocytes by the Cre-loxP system under the control of the keratin K14 promoter (Atg7Δepi mice). Autophagic activity was detected using the GFP-microtubule-associated protein light chain 3 (GFP–LC3) reporter construct and Western blot analysis of LC3. Epidermal morphology was examined by histological and ultrastructural analyses, and barrier functions were assessed by dye diffusion and water loss assays. Results Suprabasal epidermal cells of normal mice contained GFP–LC3-labeled autophagosomes and epidermal lysates of these mice showed an excess of lipidated over non-lipidated LC3. These features of active autophagy were efficiently suppressed in Atg7Δepi epidermis. Atg7Δepi mice survived the perinatal period and were apparently healthy. Histologically, their epidermis was inconspicuous and ultrastructural analysis revealed no significant defect in cornification. There was however, an increase in the thickness of corneocytes in the back skin of mutant mice. Nevertheless, resistance to dye penetration into the skin and transepidermal water loss were normal in Atg7Δepi mice. Conclusion This study demonstrates that autophagy is constitutively active in the epidermis but not essential for the barrier function of the skin.