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. 2018 Jan 23;11:7. doi: 10.3389/fnmol.2018.00007

Figure 1.

Figure 1

OxPL-evoked calcium influx is TRPA1- and TRPV1-dependent. (A) Pseudo colored images represent a calcium imaging series of adult murine DRG neurons loaded with Fura-2-AM. Calcium responses evoked by consecutive addition of PGPC (10 μM), AITC (10 μM), capsaicin (Caps, 1 μM), and KCl (50 mM) are displayed in comparison to the start situation (Basal; scale bar = 100 μm). Arrows indicate PGPC-responding neurons. (B) Relative changes in (Ca2+)i evoked by consecutive application of PGPC (10 μM), AITC (10 μM), Caps (1 μM), and KCl (50 mM) in DRG neurons. Gray traces indicate calcium responses of individual cells, ratio F(340/380). Black line: Mean of 50 cells. Colored lines (red, blue, green) represent three kinds of PGPC affected neurons. (C) Relative calcium responses of neurons activated by PGPC in the absence (PGPC) or after preincubation with antagonists specific for TRPA1 (PGPC + HC-030031) or TRPV1 (PGPC + BCTC) expressed as area under the curve, AUC, [n = 4-6 of 3 mice; mean ± SEM; one-way ANOVA Holm-Sidak; F(2, 14) = 5.83; *p < 0.028, **p < 0.008]. (D) Relative changes in (Ca2+)i of HEK293 expressing TRPA1-C3 induced by application of either OxPAPC (10 μM; mean of n = 50 cells), (E) PGPC (10 μM; mean of n = 50 cells), or (F) AITC (10 μM; mean of n = 50 cells) with consecutive addition of carvacrol (10 μM). Gray traces represent calcium responses [F(340/380)] of individual cells, black traces indicate means. (G) The relative calcium responses of HEK293 cells transfected with TRPA1-3C (TRPA1-3C) and untransfected control cells (Ctrl), stimulated with either OxPAPC, PGPC or AITC expressed as area under the curve [Ctrl; n = 5–6 of 3 cultures; data points are presented as means ± SEM; one-way ANOVA Holm-Sidak; F(2, 16) = 42.33; *p ≤ 0.05, ***p < 0.001]. (H) Indirect immunofluorescent staining of 48 h old primary DRG cultures from adult wt mice against TRPA1 (red), TRPV1 (green) and βIII-Tubulin (blue; scale bar = 100 μm).