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Immunohistochemical Localization and Multi‐gene Expression Profiling of O2 Sensor Components in Airway Chemoreceptors
Author(s) -
Pan Jie,
Yeger Herman,
Cutz Ernest
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1122.6
Subject(s) - nadph oxidase , biology , p22phox , microbiology and biotechnology , reactive oxygen species
Pulmonary neuroepithelial bodies (NEB) are airway chemoreceptors that express a membrane bound O2 sensor molecular complex (NADPH oxidase coupled to an O2 sensitive K+ channel). We used confocal immunofluorescence microscopy with specific antibodies against NADPH components and O2 sensitive K+ channels (K+O2) to label NEB in rat /rabbit lung and NEB related H146 tumor cell line. For gene expression profiling of NEB cells microdissected from human lung, and H146 cells, we used a custom MultiGene‐12TM RT‐PCR array that included NADPH oxidase complex and homologues /accessory proteins (NOX1‐4, phox‐p22, p40, p47, p67, Rac1, NOXO1 and NOXA1) and K+O2 channels (TASK1‐3, Kv ‐1.2,1.5, 2.1, 3.1, 3.3, 3.4, 4.2, 4.3). In rat lung, NOX2, NOX4, p22phox, TASK1 and Kv3.3 (and Kv3.4 in rabbit) localized mainly to the apical plasma membrane of NEB cells, and membrane or sub‐membrane regions in H146 cells. NOX2 co‐localized with p22phox, p40phox, p47phox and Kv3.3, while NOX4 co‐localized with TASK1. Gene expression profiling of NEB/H146 cells confirmed the presence of all NOX proteins except NOX3 and NOXO1, as well as all common K+O2 channels, except Kv1.5 and Kv4.3. Adjacent airway epithelial cells expressed only TASK2, Kv2.1, Kv3.4 and Kv4.2. Present findings support the plasma membrane model of O2 sensing by airway chemoreceptors that appears to be cell and organ specific. (Supported by grants from CIHR)