title: Activation of P2Y receptors causes strong and persistent shrinkage of C11-MDCK renal epithelial cells.
abstract: Purinergic receptors activate diverse signaling cascades and regulate the activity of cell volume-sensitive ion transporters. However, the effects of ATP and other agonists of P2 receptors on cell volume dynamics are only scarcely studied. In the present work, we used the recently developed dual-image surface reconstruction technique to explore the influence of purinergic agonists on cell volume in the C11-Madin-Darby canine kidney cell line resembling intercalated cells from kidney collecting ducts. Unexpectedly, we found that ATP and UTP triggered very robust (55-60%) cell shrinkage that lasted up to 2 h after agonist washout. Purinergic regulation of cell volume required increases in intracellular Ca(2+) and could be partially mimicked by the Ca(2+)-ionophore ionomycin or activation of protein kinase C by 4β-phorbol 12-myristate 13-acetate. Cell shrinkage was accompanied by strong reductions in intracellular K(+) and Cl(-) content measured using steady-state (86)Rb(+) and (36)Cl(-) distribution. Both shrinkage and ion efflux in ATP-treated cells were prevented by the anion channel blocker 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) and by the BK(Ca) channel inhibitors charybdotoxin, iberiotoxin, and paxilline. To evaluate the significance of cell-volume changes in purinergic signaling, we measured the impact of ATP on the expression of the immediate-early gene c-Fos. Thirty-minute treatment with ATP increased c-Fos immunoreactivity by approximately fivefold, an effect that was strongly inhibited by charybdotoxin and completely prevented by NPPB. Overall, our findings suggest that ATP-induced cell-volume changes are partially responsible for the physiological actions of purinergic agonists.
authors.affiliation: Research Centre, Centre Hospitalier de l’Université de Montréal (CRCHUM), Montreal, Quebec, Canada.
authors.forename: Svetlana V
authors.initials: SV
authors.lastname: Koltsova
authors.forename: Alexandra
authors.initials: A
authors.lastname: Platonova
authors.forename: Georgy V
authors.initials: GV
authors.lastname: Maksimov
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Mongin
authors.forename: Ryszard
authors.initials: R
authors.lastname: Grygorczyk
authors.forename: Sergei N
authors.initials: SN
authors.lastname: Orlov
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D020013:Calcium Signaling; D002460:Cell Line; D048429:Cell Size; D002712:Chlorides / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004285:Dogs; D004789:Enzyme Activation; D020536:Enzyme Activators / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D007476:Ionophores / Q000494:pharmacology; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000037:antagonists & inhibitors / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D026902:Potassium Channel Blockers / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D058912:Purinergic P2Y Receptor Agonists / Q000378:metabolism*; D058921:Purinergic P2Y Receptor Antagonists / Q000494:pharmacology; D058470:Receptors, Purinergic P2Y / Q000378:metabolism*; D013997:Time Factors; D014544:Uridine Triphosphate / Q000378:metabolism*
chemical_list: D002712:Chlorides; D020536:Enzyme Activators; D007476:Ionophores; D051036:Large-Conductance Calcium-Activated Potassium Channels; D026902:Potassium Channel Blockers; D016760:Proto-Oncogene Proteins c-fos; D058912:Purinergic P2Y Receptor Agonists; D058921:Purinergic P2Y Receptor Antagonists; D058470:Receptors, Purinergic P2Y; D000255:Adenosine Triphosphate; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D011188:Potassium; D014544:Uridine Triphosphate