Figure 3

Release of TRPV1 agonists. A. Twenty-four hour old rat TG cultures were pretreated with either vehicle or 30uM ketoconazole for 15 minutes following which 1 mM LA with vehicle or the same concentration of ketoconazole was applied to the neurons for 2 minutes and calcium accumulated was tested by measuring ratio of fluorescence produced at 340 and 380 nm wavelengths. The experiment was performed with three independent neuronal cultures and mean responses for each group is plotted. Data were analyzed using 2-tailed student’s t-test. Error Bars: S.E.M. Noise indicates basal fluorescence ratio 340/360 with application of Hanks Buffer. B. Lipophilic extracts were made from inflamed skin from rats, pretreated with vehicle or 18 mM ketoconazole in vivo as well as vehicle or 150uM ketoconazole ex vivo and applied onto 5-day old rat TG cultures for 15 minutes. Neurons were pretreated with either vehicle or 1uM I-RTX for 15 minutes prior to extract application. The superfusates were collected and subjected to radioimmunoassay for estimation of CGRP release. Three independent experiments were performed and data plotted is the mean of % release from baseline for each group. Data were normalized such that baseline release from neurons was made to be 100%. Data were analyzed using one-way ANOVA and Neuman-Keuls post hoc test. Error Bars: S.E.M.