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Morphological, molecular and functional aspects of the interaction between the P2X7 receptor and pannexin-1 in the enteric glial cells following intestinal ischemia/reperfusion

Grant number: 14/25927-2
Support type:Regular Research Grants
Duration: October 01, 2015 - March 31, 2018
Field of knowledge:Biological Sciences - Morphology - Anatomy
Principal researcher:Patricia Castelucci
Grantee:Patricia Castelucci
Home Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Enteric glial cells are considered as a structural support for enteric neurons, but recent evidence suggests an interaction glia-neuron communication contributes to neuronal maintenance and that these changes (functional or morphological) cells may play a role in gastrointestinal disorders. Ischemia and reperfusion are severe clinical events with high mortality occurring in transplantation, intestinal blockage and embolism which consequently affect the enteric neurons and glial cells. ATP is involved in the regulation of various pathophysiological processes, can be co-released with other neurotransmitters and their extracellular signaling may occur via P2X and P2Y receptor family. Some characteristics have been described for pannexin-1 and mediate the purinergic signaling in the nervous system by signaling ionotropic and metabotropic receptors. The P2X7 receptor has a role in various intracellular events and acts together with pannexin-1. This design aims to study the effects of ischemia / reperfusion (I / R) enteric glial cells in the ileum of rats that express the P2X7 receptor and pannexin-1. Intestinal ischemia is obtained by obstruction of blood flow in vessels of the ileal period of 45 minutes, followed by reperfusion for 24 hours, 14 days and 28 days. In sham occlusion of the ileal vessels will not occur. The antagonists of P2X7 receptor (Brilliant Blue G/BBG) and pannexin-1 (Probenecid)- will be used following I / R. The tissues will be prepared for immunohistochemistry methods of double / triple markings P2X7 receptor, S100B (glial marker) or GFAP (glial marker) and Hu (neuronal marker ) and pannexin-1. The qualitative and quantitative analysis will be made with a fluorescence microscope and confocal laser scanning. Also, analysis of ultrastructural, protein expression by western blotting (P2X7 receptor, pannexin-1, S100B/GFAP and Hu) and gene expression will be made. Furthermore, the involvement of signaling pathways of the proteins will be studied mitogen-activated kinases (MAPK) ERK-1/2, p38 MAPK, NF-KB and NFAT and cell death as caspase 3 enzyme immunoassays will be performed cytokine IL-1B, IL-6, IL-10 and TNFalfa and analysis of the functional aspects of intestinal motility and in sham I/R. Understanding the role of enteric glia and interactions with P2X7 receptor pannexin-1 may contribute to therapeutic strategies for gastrointestinal disorders caused by ischemia and reperfusion. (AU)

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Scientific publications (4)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
AGUIAR, BRUNA ANDRADE; ORECHIO, DAILIANY; FRATINI, PAULA; OLIVEIRA CARREIRA, ANA CLAUDIA; CASTELUCCI, PATRICIA; MIGLINO, MARIA ANGELICA. Isolation and Characterization of Pancreatic Canine Fetal Cells at the Final Stage of Gestation. Anatomical Record-Advances in Integrative Anatomy and Evolutionary Biology, v. 302, n. 8, SI, p. 1409-1418, AUG 2019. Web of Science Citations: 0.
PALOMBIT, KELLY; MENDES, CRISTINA EUSEBIO; TAVARES-DE-LIMA, WOTHAN; BARRETO-CHAVES, MARIA LUIZA; CASTELUCCI, PATRICIA. Blockage of the P2X7 Receptor Attenuates Harmful Changes Produced by Ischemia and Reperfusion in the Myenteric Plexus. Digestive Diseases and Sciences, v. 64, n. 7, p. 1815-1829, JUL 2019. Web of Science Citations: 0.
MENDES, CRISTINA EUSEBIO; PALOMBIT, KELLY; TAVARES-DE-LIMA, WOTHAN; CASTELUCCI, PATRICIA. Enteric glial cells immunoreactive for P2X7 receptor are affected in the ileum following ischemia and reperfusion. ACTA HISTOCHEMICA, v. 121, n. 6, p. 665-679, 2019. Web of Science Citations: 0.
ORECHIO, DAILIANY; AGUIAR, BRUNA ANDRADE; DINIZ, GIOVANNE BARONI; BITTENCOURT, JACKSON CIONI; HAEMMERLE, CARLOS A. S.; WATANABE, II-SEI; MIGLINO, MARIA ANGELICA; CASTELUCCI, PATRICIA. Morphological and Cellular Characterization of the Fetal Canine (Canis lupus familiaris) Subventricular Zone, Rostral Migratory Stream, and Olfactory Bulb. Anatomical Record-Advances in Integrative Anatomy and Evolutionary Biology, v. 301, n. 9, SI, p. 1570-1584, SEP 2018. Web of Science Citations: 1.

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