Epidermal neurobiology: exploiting the endocannabinoid signaling.

  • Hachem, Jean-Pierre, (Co-Promotor)
  • Roseeuw, Diane (Administrative Promotor)

Project Details

Description

C- Methods and objectives: In Vivo models: knockout animals Skin stress models: Murine epidermis will be subject to 4 types of stress models: Acute Barrier stress: by tape stripping of the SC. Removal of the later will induce a prompt response characterized by the formation of new corneocytes including: cytokine release, instant secretion of lamellar bodies of the underlying epidermis, the activation of terminal differentiation of the suprabasal cell layers and a proliferative reponse of the basal cell layers. Chronic barrier stress: bi-daily tape stripping of the SC corneum can further produce epidermal hyperplasia, mimicking hyperproliferative skin diseases such as psoriasis.Irritant and contact dermatitis: to elicit skin inflammation. Knockout mouse models to address Endocannabinoid: At least 4 types of mouse models to specifically address all the above mentioned pathways. CB1 ko mice have been generated within the laboratory of Catherine Ledent (ULB, Erasme). CB2 and TRPV1 ko animals are available at Jackson laboratories and CB2 ko are already breeding at the VUB facility. Double CB2-CB1 ko are being generated from the respective colonies at the ULB. PPAR-alpha deficient animals are available at the VA medical center, University of California San Francisco, longtime collaborator with our research unit (see table 1 one for animal origin details). . Aim 1: To Assess the Relationship Between EC signaling and SC Barrier Function/Lipid (Ceramide) Content/Epidermal Differentiation. To assess how EC signaling pathway contributes to the permeability barrier function. Both ko an wt animals will be subject to acute barrier stress and chronic barrier stress as described above. Functional assessment will be performed using the following parameters as a comprehensive characterization of epidermal function: Transepidermal water loss (TEWL), a measurement of cutaneous permeability barrier function, is measured with a TEWAmeter (Courage and Khazaka, Cologne, Germany). Barrier recovery is determined following repeated applications of cellophane tape (Scotch type, 3M) until TEWL reaches 6-8 mg per cm2 per h. Barrier recovery is determined by measuring TEWL at 3 and 6 h following barrier disruption. SC integrity is defined as the number of tape strippings required to produce defined elevations in TEWL, and cohesion of the stratum corneum as the amount of protein removed with each stripping. The protein content per stripping is measured with the Bio-Rad Assay Kit (Hercules, CA), using bovine plasma ?-globulin as the standard. Surface pH is measured with a flat glass electrode from Mettler-Toledo (Giessen, Germany), attached to a pH meter (Skin pH Meter pH 900, Courage and Khazaka, Cologne, Germany). To address the morphological basis in the observed variation in permeability function skin biopsies from the both ko and wt animals will be subject to Electron Microscopy analysis. Samples are minced to
AcronymFWOAL495
StatusFinished
Effective start/end date1/01/0931/12/12

Keywords

  • Dermatology

Flemish discipline codes

  • Basic sciences