P2Y2 receptor promotes intestinal microtubule stabilization and mucosal re-epithelization in experimental colitis

• Published in: Journal of cellular physiology Vol. 228; no. 1; pp. 99 - 109
• Main Authors: Degagné, Émilie, Degrandmaison, Jade, Grbic, Djordje M., Vinette, Valérie, Arguin, Guillaume, Gendron, Fernand-Pierre
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.01.2013; Wiley Subscription Services, Inc
• Subjects: Adenosine Triphosphate - pharmacology; Animals; Calcium - metabolism; Cell Line; Cell Movement - drug effects; Cell Movement - physiology; Colitis - chemically induced; Colitis - metabolism; Epithelial Cells - metabolism; Glycogen Synthase Kinase 3 - genetics; Glycogen Synthase Kinase 3 - metabolism; Glycogen Synthase Kinase 3 beta; Integrin alphaV - genetics; Integrin alphaV - metabolism; Intestinal Mucosa - cytology; Intestinal Mucosa - drug effects; Intestinal Mucosa - metabolism; Mice; Microtubules - metabolism; Phosphatidylinositol 3-Kinases - genetics; Phosphatidylinositol 3-Kinases - metabolism; Proto-Oncogene Proteins c-akt - genetics; Proto-Oncogene Proteins c-akt - metabolism; Purinergic P2Y Receptor Agonists - pharmacology; Rats; Receptors, Purinergic P2Y2 - genetics; Receptors, Purinergic P2Y2 - metabolism; Sodium sulfate; Tubulin - genetics; Tubulin - immunology; Tubulin - metabolism; Uridine Triphosphate - pharmacology; Wound Healing - drug effects
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.24109

P2Y2 receptor expression is increased in intestinal epithelial cells (IECs) during inflammatory bowel diseases (IBDs). In this context, P2Y2 stimulates PGE2 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 P2Y2 agonist 2‐thioUTP on remission. We showed that P2Y2 increased cell migration and wound closure by recruiting Go protein with the cooperation of integrin αv. Following P2Y2 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, P2Y2 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. J. Cell. Physiol. 228: 99–109, 2013. © 2012 Wiley Periodicals, Inc.