Fig. 3. P2Y2 receptor activation elicits CysLTs from airway BrCs.
(A) Transcripts of differentially expressed GPCRs in olfactory nasal BrCs compared to nasal EpCs using criteria of log2 fold change of 2 or greater expression in one relative to the other with a p adjusted value of <0.01. (B) Transcripts for genes encoding purinergic receptors expressed at levels of 5 or above among nasal BrCs. (C, E) BrCs (EpCAM+CD45low/−eGFP+) were isolated from the nasal mucosa (nose) of ChAT-eGFP mice and were stimulated ex vivo with the indicated doses of ATPγS (C) or UTP (E) for 30 min. Where indicated, BrCs were pre-treated for 15 min with the P2RY2-specific inhibitor AR-C118925, the FLAP inhibitor MK-886 or HBSS and subsequently stimulated with ATPγS. (D) Mouse BMMCs were stimulated with ATPγS for 30 min. The concentrations of CysLTs in the supernatants were measured by enzyme immunoassay at 30 min. (F) ChAT-eGFP+ BrCs were pre-treated for 15 min with the P2X4-specific inhibitor 5-BDBD and subsequently stimulated with ATPγS. CysLTs were measured in the supernatant at 30 min. (G) CD45lowEpCAMhi BrCs from WT, Ltc4s−/− and P2ry2−/− mice were stimulated ex vivo with 0.1 mM of ATPγS. The concentration of CysLTs was measured by enzyme immunoassay. (H, I) ATP at the indicated dose was administered intranasally to naïve WT, Pou2f3−/− (H) and P2ry2−/− (I) mice and nasal lavage (NL) was obtained at 30 min. The concentration of CysLTs were measured by ELISA after acetone precipitation. Data are means ± SEM, each dot is a separate biological replicate, data are pooled from at least 2 independent experiments * p<0.05, ** p<0.01, *** p<0.001.