title: Permeability barrier requirements regulate epidermal beta-glucocerebrosidase.
abstract: The intercellular spaces of the outermost layers of the epidermis (stratum corneum, SC) of terrestrial mammals contain a mixture of lipids, enriched in ceramides that are critical for the epidermal permeability barrier. Whereas glucosylceramides (GlcCer) are synthesized in abundance in the epidermis, they disappear coincident with an increase of ceramides (Cer) in the SC. Hence, hydrolysis of GlcCer to Cer by beta-glucocerebrosidase (GlcCer'ase), may be required for permeability barrier homeostasis. We determined first whether modulations in epidermal GlcCer'ase activity and mRNA levels occur in response to barrier disruption; and second, how GlcCer'ase inhibitors influence barrier function and SC membrane ultrastructure. Barrier disruption significantly increased epidermal GlcCer'ase mRNA levels, with a 2.8-fold increase over untreated control levels at 8 h (P < 0.01). GlcCer'ase activity was increased in whole epidermis (34%; P < 0.02) 24 h after barrier disruption. Localization of GlcCer'ase activity showed an increase (33%; P < 0.05) in the outer epidermis (SC and stratum granulosum), without a change in lower epidermal activity (stratum spinosum and stratum basale). Furthermore, a single topical application of the GlcCer'ase inhibitor, bromoconduritol-B-epoxide (BrCBE), inhibited enzyme activity (98%) and significantly delayed permeability barrier recovery after acetone treatment. In addition, BrCBE treatment disrupted SC intercellular lamellar bilayers, without evidence of cellular toxicity. These results indicate that epidermal processing of GlcCer to Cer by GlcCer'ase is required for barrier homeostasis, and that this important enzymatic step is regulated by barrier requirements.
authors.affiliation: Department of Dermatology, University of California School of Medicine, San Francisco.
authors.forename: W M
authors.initials: WM
authors.lastname: Holleran
authors.forename: Y
authors.initials: Y
authors.lastname: Takagi
authors.forename: G K
authors.initials: GK
authors.lastname: Menon
authors.forename: S M
authors.initials: SM
authors.lastname: Jackson
authors.forename: J M
authors.initials: JM
authors.lastname: Lee
authors.forename: K R
authors.initials: KR
authors.lastname: Feingold
authors.forename: P M
authors.initials: PM
authors.lastname: Elias
mesh_terms: D000096:Acetone; D000818:Animals; D002240:Carbohydrate Sequence; D004817:Epidermis / Q000187:drug effects / Q000201:enzymology*; D004852:Epoxy Compounds / Q000494:pharmacology; D005962:Glucosylceramidase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006706:Homeostasis / Q000187:drug effects; D007294:Inositol / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D051379:Mice; D008812:Mice, Hairless; D008969:Molecular Sequence Data; D010539:Permeability / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D004852:Epoxy Compounds; D012333:RNA, Messenger; D000096:Acetone; C089157:bromoconduritol-B-epoxide; D007294:Inositol; D005962:Glucosylceramidase