title: P2Y2 receptor promotes intestinal microtubule stabilization and mucosal re-epithelization in experimental colitis.
abstract: P2Y(2) receptor expression is increased in intestinal epithelial cells (IECs) during inflammatory bowel diseases (IBDs). In this context, P2Y(2) stimulates PGE(2) release by IECs, suggesting a role in wound healing. For this study, we have used the non-cancerous IEC-6 cell line. IEC-6 cell migration was determined using Boyden chambers and the single-edged razor blade model of wounding. The receptor was activated using ATP, UTP, or 2-thioUTP. Pharmacological inhibitors, a blocking peptide, a neutralizing antibody and interfering RNAs were used to characterize the signaling events. Focal adhesions and microtubule (MT) dynamics were determined by immunofluorescence using anti-vinculin and anti-acetylated-α-tubulin antibodies, respectively. In vivo, the dextran sodium sulfate mouse model of colitis was used to characterize the effects of P2Y(2) agonist 2-thioUTP on remission. We showed that P2Y(2) increased cell migration and wound closure by recruiting Go protein with the cooperation of integrin α(v) . Following P2Y(2) activation, we demonstrated that GSK3β activity was inhibited in response to Akt activation. This leads to MT stabilization and increased number of focal adhesions. In vivo, P2Y(2) activation stimulates remission, as illustrated by a reduction in the disease activity index values and histological scores as compared to control mice. These findings highlight a novel function for this receptor in IECs. They also illustrate that P2Y receptors could be targeted for the development of innovative therapies for the treatment of IBDs.
authors.affiliation: Department of Anatomy and Cell Biology, Université de Sherbrooke, Sherbrooke, Quebec, Canada J1H 5N4.
authors.forename: Emilie
authors.initials: E
authors.lastname: Degagné
authors.forename: Jade
authors.initials: J
authors.lastname: Degrandmaison
authors.forename: Djordje M
authors.initials: DM
authors.lastname: Grbic
authors.forename: Valérie
authors.initials: V
authors.lastname: Vinette
authors.forename: Guillaume
authors.initials: G
authors.lastname: Arguin
authors.forename: Fernand-Pierre
authors.initials: FP
authors.lastname: Gendron
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D003092:Colitis / Q000139:chemically induced / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism*; D038362:Glycogen Synthase Kinase 3 / Q000235:genetics / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D039564:Integrin alphaV / Q000235:genetics / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008870:Microtubules / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D058912:Purinergic P2Y Receptor Agonists / Q000494:pharmacology; D051381:Rats; D058473:Receptors, Purinergic P2Y2 / Q000235:genetics / Q000378:metabolism*; D014404:Tubulin / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014544:Uridine Triphosphate / Q000494:pharmacology; D014945:Wound Healing / Q000187:drug effects
chemical_list: D039564:Integrin alphaV; D058912:Purinergic P2Y Receptor Agonists; D058473:Receptors, Purinergic P2Y2; D014404:Tubulin; D000255:Adenosine Triphosphate; D019869:Phosphatidylinositol 3-Kinases; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; C000605508:Gsk3b protein, rat; D051057:Proto-Oncogene Proteins c-akt; D038362:Glycogen Synthase Kinase 3; D002118:Calcium; D014544:Uridine Triphosphate