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Purinergic signaling in the pulmonary neuroepithelial body microenvironment unraveled by live cell imaging

De Proost, I., Pintelon, I., Wilkinson, William James, Goethals, S., Brouns, I., Van Nassauw, L., Riccardi, Daniela, Timmermans, J. P., Kemp, Paul J and Adriaensen, D. 2009. Purinergic signaling in the pulmonary neuroepithelial body microenvironment unraveled by live cell imaging. The FASEB Journal 23 (4) , pp. 1153-1160. 10.1096/fj.08-109579

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Abstract

Pulmonary neuroepithelial bodies (NEBs) are densely innervated groups of complex sensory airway receptors involved in the regulation of breathing. Together with their surrounding Clara-like cells, they exhibit stem cell potential through their capacity to regenerate depopulated areas of the epithelium following lung injury. We have employed confocal live cell imaging microscopy and novel electrophysiological techniques in a new ex vivo lung slice model to unravel potential purinergic signaling pathways within the NEB microenvironment. Quinacrine histochemistry indicated high amounts of vesicular ATP in NEB cells. Using a “reporter-patching” method adapted to create a uniquely sensitive and selective biosensor for the direct detection of ATP release from NEBs ex vivo, we demonstrated quantal ATP release from NEBs following their depolarization. Enhancing enzymatic extracellular ATP hydrolysis or inhibiting P2 receptors confirmed the central role of ATP in paracrine interactions between NEB cells and Clara-like cells. Combined calcium imaging, pharmacology, and immunohistochemistry showed that ligand-binding to functional P2Y2 receptors underpins the activation of Clara-like cells. Hence, NEB cells communicate with their cellular neighbors in the NEB microenvironment by releasing ATP, which rapidly evokes purinergic activation of surrounding Clara-like cells. Besides ATP acting on the P2X3 receptor expressing vagal sensory nerve terminals between NEB cells, local paracrine purinergic signaling within this potential stem cell niche may be important to both normal airway function, airway epithelial regeneration after injury, and/or the pathogenesis of small cell lung carcinomas.—De Proost, I., Pintelon, I., Wilkinson, W. J., Goethals, S., Brouns, I., Van Nassauw, L., Riccardi, D., Timmermans, J.-P., Kemp, P. J., Adriaensen, D. Purinergic signaling in the pulmonary neuroepithelial body microenvironment unraveled by live cell imaging.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Publisher: Federation of American Society of Experimental Biology
ISSN: 0892-6638
Last Modified: 04 Jun 2017 02:06
URI: http://orca-mwe.cf.ac.uk/id/eprint/9112

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