Fig. 1 | Nature Communications

Fig. 1

From: Preventing acute asthmatic symptoms by targeting a neuronal mechanism involving carotid body lysophosphatidic acid receptors

Fig. 1

LPA stimulates the carotid body. a RT-PCR: LPAr (L) 1, 3, 4, and 6 in carotid body (left), LPAr 3 and 6 in petrosal ganglia (center), and LPAr 1, 3, 4, and 6 in superior cervical ganglia (right) in reference to a housekeeping gene, hypoxyribosyltransferase (H; cDNA ladder, Ld, was used to identify tissues used for each gel, 100 bp marker indicated). b Calcium imaging with Fura-2 loaded carotid body glomus cells (12 cells on 5 cover slips) reveals an increase of intracellular calcium in response to 5 µM LPA (18:1 unless otherwise stated) and 20 mM K+. F2,35 (one-way ANOVA) = 18.220, p < 0.001; Holm–Šidák post hoc: difference from baseline, ***p < 0.01. c En bloc perfused carotid body preparation used to record chemosensory afferents in the carotid sinus nerve (CSN). d LPA 18:1 causes an increase in CSN activity. e The dose-dependent relationship differed slightly between several species of LPA found in blood (16:0, 18:1, and 18:2); F4,53 (two-way ANOVA: species × dose) = 3.380, p = 0.021. Holm–Šidák post hoc: all species 5 and 10 µM different from 2.5 µM, ***p < 0.001. f Effects of TRPV1 and LPAr antagonists on CSN response to LPA. No blockade (black trace), TRPV1 blockade (AMG9810 10 µM, teal trace, or LPAr blockade (BrP-LPA 1.5 µM, mauve trace; Ki16425 5 µM, fuscia trace). g Summary data. Responses 2.5 µM LPA: F3,23 (one-way ANOVA) = 5.031, **p = 0.01. Holm–Šidák post hoc: LPA different from BrP-LPA (p = 0.026) and Ki16425 (p = 0.018). Responses 5 µM: F3,23 (one-way ANOVA) = 24.547, ***p < 0.001. Holm–Šidák post hoc: LPA different from BrP-LPA, Ki16425, and AMG9810 (p < 0.001); AMG9810 different from Brp-LPA (p = 0.018) and Ki16425 (p = 0.044); Responses 10 µM: F3,23 (one-way ANOVA) = 14.231, ***p < 0.001. Holm–Šidák post hoc: LPA different from BrP-LPA (p < 0.001); Ki16425 (p < 0.001); AMG9810 (p = 0.022); AMG9810 different from BrP-LPA (p = 0.022). h Summary data of 5 µM LPA (blue), with TRPV1 blockade (AMG9810, 10 µM; teal), LPAr blockade (BrP-LPA, 1.5 µM; mauve; Ki16425, 5 µM; fuscia), or LPAr + TRPV1 blockade (AMG9810 + BrPLPA red; AMG9810 + Ki16425 orange), F5,35 (one-way ANOVA) = 26.164, p < 0.001. Holm–Šidák post hoc: AMG9810 + LPA significantly different from Ki16425 + AMG9810 + LPA, p = 0.005; LPA significantly different from all other groups, ***p < 0.001. All data are presented as mean ± sem

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