Figure 1.

Olfactory receptors (ORs) expressed in cultured preparations of human airway epithelial cells. (A) OR gene expression by microarray analysis of mRNAs isolated from primary airway cell preparations. Shown is the mean (± SEM) from 10 independent donors. (B) Representative photomicrograph of immunostaining for the OR, OR2H3 (green), in primary-culture human airway cells demonstrating an interdigitating morphology. Cilia are labeled in red (acetylated α-tubulin, open arrowhead). Images are confocal z-stacks. (C) Colocalization of odor receptors, OR2W1 (green) and OR2F1 (red), in human airway preparations. Images are confocal z-stacks. (D) Protein blot analyses. OR2F1 and OR2W1 are expressed in human primary-culture airway cells. Each lane represents a sample from a single donor. The expected size of the corresponding ORs is indicated (∼ 45 kD; closed arrows). (E) In situ hybridization of human airway tissue sections using an OR2W1-specific riboprobe. (E¹) antisense probe; (E²) enlargement of box in (E¹); (E³) sense control riboprobe. Dotted lines represent the basal aspect of the stratified epithelium. Both sense and antisense probes were hybridized to sequential sections of the same tissue. (F) Cultured cells immunostained for 5-HT show extensive interdigitating morphology. Upper panel is a confocal x,y section. Lower panel is a z,y section (“stack”) of the upper panel. Note that some of the cellular extensions show projections to the apical side (white arrowheads). Nuclei are labeled with 4′,6-diamidino-2-phenylindole (blue). Scale bars, 50 μm.