Epiplakin Is Dispensable for Skin Barrier Function and for Integrity of Keratin Network Cytoarchitecture in Simple and Stratified Epithelia

Abstract

Epiplakin, a giant epithelial protein of >700 kDa, belongs to the plakin family of cytolinker proteins. It represents an atypical family member, however, as it consists entirely of plakin repeat domains but lacks any of the other domains commonly shared by plakins. Hence, its putative function as a cytolinker protein remains to be shown. To investigate epiplakin's biological role, we generated epiplakin-deficient mice by gene targeting in embryonic stem cells. Epiplakin-deficient mice were viable and fertile, without developing any discernible phenotype. Ultrastructurally, their epidermis revealed no differences compared to wild-type littermates, and cornified envelopes isolated from skin showed no alterations in shape or stability. Furthermore, neither embryonal formation nor later function of the epithelial barrier was affected. In primary cultures of epiplakin-deficient keratinocytes, the organization of actin filaments, microtubules, and keratin networks was found to be normal. Similarly, no alterations in keratin network organization were observed in simple epithelia of small intestine and liver or in primary hepatocytes. We conclude that, despite epiplakin's abundant and highly specific expression in stratified and simple epithelia, its absence in mice does not lead to severe skin dysfunctions, nor has it detectable consequences for keratin filament organization and cytoarchitecture of cells.

We thank Manfred Kraus (University of Cologne, Germany) for the neomycin resistance cassette and Erwin Wagner (IMP, Vienna, Austria) for the herpes simplex virus thymidine kinase cassette. We thank Takashi Hashimoto (Kurume University School of Medicine, Japan), Kurt Zatloukal (Medical University, Graz, Austria), Fiona Watt (Imperial Cancer Research Fund, United Kingdom), and Dennis Roop (Baylor College of Medicine, TX) for generous gifts of antibodies. The help of Fritz Zschiegner (Medical University of Innsbruck, Austria) in anesthetizing mice is greatly appreciated. We also thank Zelkja Stojanovic and Ulrike Köck for dedicated technical assistance in generating and analyzing knockout mice.

This work was supported by grant F006-11 from the Austrian Science Research Fund.