title: Functional expression of the damage-associated molecular pattern receptor P2X7 on canine kidney epithelial cells.
abstract: Epithelial cells are important in inflammation and immunity. In this study, we examined if Madin-Darby canine kidney (MDCK) epithelial cells express functional P2X7 receptors, which bind the damage-associated molecular pattern extracellular adenosine 5'-triphosphate (ATP). Reverse transcription (RT)-PCR and immunoblotting revealed the expression of P2X7 in MDCK cells. A flow cytometric assay demonstrated that ATP or 2'(3')-O-(4-benzoylbenzoyl)ATP induced ethidium(+) uptake into MDCK cells, and that this process was impaired by the P2X7 antagonists KN-62 and A438079. RT-PCR also demonstrated the presence of Toll-like receptor 4, NALP3, caspase-1, interleukin-1β and interleukin-18 in MDCK cells, as well as in positive control LPS-primed canine monocytes. In conclusion, the MDCK epithelial cell line expresses functional P2X7, as well as Toll-like receptor 4 and molecules associated with the NALP3 inflammasome. This cell line may help elucidate the role of these molecules in kidney epithelial cells and renal disorders in dogs and humans.
authors.affiliation: School of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, Australia.
authors.forename: Iman
authors.initials: I
authors.lastname: Jalilian
authors.forename: Mari
authors.initials: M
authors.lastname: Spildrejorde
authors.forename: Aine
authors.initials: A
authors.lastname: Seavers
authors.forename: Belinda L
authors.initials: BL
authors.lastname: Curtis
authors.forename: Jason D
authors.initials: JD
authors.lastname: McArthur
authors.forename: Ronald
authors.initials: R
authors.lastname: Sluyter
mesh_terms: D000818:Animals; D015415:Biomarkers; D002460:Cell Line; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000276:immunology*; D058847:Inflammasomes / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D058486:Receptors, Purinergic P2X7 / Q000235:genetics / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism
chemical_list: D015415:Biomarkers; D058847:Inflammasomes; D012333:RNA, Messenger; D058486:Receptors, Purinergic P2X7; D051197:Toll-Like Receptor 4