CGRP release assay from skin explants and cultured neurons

Skin punch biopsies (10 mm) were collected from the nape of the murine psoriasis model and rapidly transferred into 24-well plates containing 1 mL of Dulbecco's modified Eagle medium. Explants were incubated at 38°C for 30 min. After incubation, the supernatant from the organ cultures was collected and assayed to measure CGRP concentrations with the CGRP EIA kit (cat. no. 589001, Cayman Chemical) according to the manufacturer`s instructions ⁴³. The same kit was used to determine the amount of CGRP released into the media of cultured murine and human sensory neurons stimulated by capsaicin (1 µM, 30 min) with or without RvD3 (100 nM or 1 µg). The same procedure described in section 2.8. was used to obtain murine cultured sensory neurons, whereas human cultured sensory neurons were prepared as previously described ⁴⁴.