Analysis of purine receptor expression and functionality in alveolar epithelial cells

Despite its fundamental role in providing an extensive surface for gas exchange, the alveolar epithelium (AE) serves as an immunological barrier through, e.g., the release of proinflammatory cytokines and secretion of surfactant to prevent alveolar collapse. Thus, AE is important for sustaining lung...

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Bibliographic Details
Published in:Purinergic signalling 2020-06, Vol.16 (2), p.213-229
Main Authors: Olotu, Cynthia, Kiefmann, Martina, Ronneburg, Cornelia, Lehmensiek, Felix, Cuvenhaus, Annelie, Meidl, Volker, Goetz, Alwin E., Kiefmann, Rainer
Format: Article
Language:English
Subjects:
Alveoli
Biomedical and Life Sciences
Biomedicine
Calcium
Cancer Research
Cytokines
Epithelial cells
Epithelium
Gas exchange
Homeostasis
Human Physiology
Inflammation
mRNA
Neurosciences
Original
Original Article
Pharmacology/Toxicology
Purine receptors
siRNA
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Summary:Despite its fundamental role in providing an extensive surface for gas exchange, the alveolar epithelium (AE) serves as an immunological barrier through, e.g., the release of proinflammatory cytokines and secretion of surfactant to prevent alveolar collapse. Thus, AE is important for sustaining lung homeostasis. Extracellular ATP secreted by alveolar epithelial cells (AECs) is involved in physiological and pathological conditions and acts mainly through the activation of purine receptors (P2Rs). When studying P2R-mediated processes, primary isolated type II AECs (piAECs) still represent the gold standard in in vitro research, although their preparation is time-consuming and requires the sacrifice of many animals. Hence, cultivated immortalized and tumor-derived AEC lines may constitute a valuable alternative. In this work, we examined P2R expression and functionality in piAECs, in immortalized and tumor-derived AEC lines with the purpose of gaining a better understanding of purinergic signaling in different cell systems and assisting researchers in the choice of a suitable cell line with a certain P2R in demand. We combined mRNA and protein analysis to evaluate the expression of P2R. For pharmacological testing, we conducted calcium ([Ca 2+ ]) measurements and siRNA receptor knockdown. Interestingly, the mRNA and protein levels of P2Y 2 , P2Y 6, and P2X 4 were detected on all cell lines. Concerning functionality, P2XR could be narrowed to L2 and piAECs while P2YR were active in all cell lines.
ISSN:1573-9538
1573-9546
DOI:10.1007/s11302-020-09696-0