Fig. 7
Modulation of TRPA1 channels by Sema4C/Plexin-B2/RhoA-ROCK signaling. a Modulation of calcium transients evoked in DRG sensory neurons by AITC (50 µM) by pretreatment with either vehicle or Sema4C (150 nM) in the presence or absence of ROCK inhibitor (50 μg/ml); n = 3 independent culture experiments. Note that AITC was applied together with Sema4C or vehicle treatment and again 30 min later alone - the responses elicited by a second application of AITC are shown in a. b Sema4C-induced changes in subcellular localization of myc-tagged TRPA1 in cultured DRG neurons. Shown are typical examples, line profiles on confocal images and quantification of fluorescence intensity at the cell membrane relative to cytoplasmic localization (n = 40 cells/group for Sema4C treatment and n = 43 for vehicle treatment). Scale bar, 25 µm. c Nocifensive responses to intraplantar application of Allyl isothiocyanate (AITC, 3.5 mM), a TRPA1 agonist, 3–4 h following intraplantar injection of vehicle or Sema4C (n = 5 mice/group for vehicle treatment, n = 8 for Sema4C treatment). d, e Nocifensive responses to intraplantar application of AITC following intraplantar injection of vehicle or Sema4C in mice lacking Plexin-B2 conditionally in adult DRG (AAV-DRG-PB2−/−) and their corresponding AAV-DRG-PB2fl/fl controls (n = 8 mice/group), d or in mice lacking Sema4C (Sema4C−/−) and their wild-type littermates (n = 6 mice/group), e. ANOVA for random measures followed by Tukey’s test was performed in a, d and e, Student’s t-test was performed in b and c. In all panels, P < 0.05 indicated by * as compared to the corresponding control groups and by † as compared to basal and by # as compared to Sema4C-treated group. Error bars represent s.e.m
