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Study of the chemical coding and P2X2 and P2X7 purina receptors of the ileum enteric nervous system of rats submitted to ischemia/reperfusion intestinaL of the ileal artery

Grant number: 08/05314-5
Support type:Regular Research Grants
Duration: April 01, 2009 - March 31, 2011
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

The ATP (adenosine 5-triphosphate) is have known as a neurotransmitter and their receptor are the family of P2X1-7 (ABBRACHIO AND BURNSTOCK, 1998) and have shown the presence of purine P2X2-3 receptor in the enteric nervous system (CASTELUCCI et al., 2002, POOLE et al., 2002; CASTELUCCI et al., 2003). In the gastrointestinal tract, the ischemia /reperfusion intestinal (I/R-i) causes morphological changes in enteric neurons (LINDESTRÖM and EKBLAD, 2004).The I/R-i is observed in acute ischemia mesenteric and Crohn's disease presenting as processes like necrosis of the bowel. This project aims to study the ileum enteric nervous system of animals subjected to I/R-i of the ileal artery: a) the chemical coding of enteric neurons immunoreactivy to nitric oxide synthase (NOS), calbindin, calretinin, choline acetyl transferase (ChAT), VIP (vase intestinal peptide active), NPY (Y neuropeptide) coexpression with neuronal marker with anti-Huc/HuD (pan neuronal) and with P2X2 and P2X7 receptor; b) the expression of the P2X2 and P2X7 receptor in the enteric nervos system c) the profile of neurons and neuronal density ( neuron/mm2) of P2X2-ir(immunoreactivity) and P2X7-ir receptor neurons, calbindin-ir neurons, calretinin-ir neurons, CHAT-ir neurons, the VIP-ir and NPY-ir d) the process of apoptosis and necrosis of enteric neurons and e) enteric glial cell. Rats ileum will be analysed (n=4 in ischemic and controls=4) subjected to ischemia for 30-40 minutes and 2, 12, 24, 72 hours, 7 days and 1 month of reperfusion. Tissues will be prepared by immunohistochemical methods of simple and double labelling of the P2X2 and P2X7 receptor and the neuronal marker (anti-HuC/HuD), calbindin, calretinin, ChAT, VIP, NPY. The qualitative and quantitative analysis of counts of colocalizations, densities of neuronal and neuronal profiles will be obtained from the fluorescence microscopes and Confocal scanning of the laser. There will be examining the detection of protein of the P2X2 and P2X7 receptors. Statistical analyses will be done. (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)
SCHAFER, BARBARA TAVARES; SILVEIRA, MARIANA POVOA; PALOMBIT, KELLY; MENDES, CRISTINA EUSEBIO; WATANABE, II SEI; MIGLINO, MARIA ANGELICA; CASTELUCCI, PATRICIA. Morphological Characterization of the Myenteric Plexus of the Ileum and Distal colon of Dogs Affected by Muscular Dystrophy. Anatomical Record-Advances in Integrative Anatomy and Evolutionary Biology, v. 301, n. 4, p. 673-685, . (08/05314-5, 12/00259-1)
PALOMBIT, KELLY; MENDES, CRISTINA EUSEBIO; TAVARES-DE-LIMA, WOTHAN; SILVEIRA, MARIANA POVOA; CASTELUCCI, PATRICIA. Effects of Ischemia and Reperfusion on Subpopulations of Rat Enteric Neurons Expressing the P2X7 Receptor. Digestive Diseases and Sciences, v. 58, n. 12, p. 3429-3439, . (08/05314-5)
DA SILVA, MARCOS VINICIUS; MAROSTI, ALINE ROSA; MENDES, CRISTINA EUSEBIO; PALOMBIT, KELLY; CASTELUCCI, PATRICIA. Submucosal neurons and enteric glial cells expressing the P2X7 receptor in rat experimental colitis. ACTA HISTOCHEMICA, v. 119, n. 5, p. 481-494, . (08/05314-5, 12/00259-1)
MENDES, CRISTINA EUSEBIO; PALOMBIT, KELLY; VIEIRA, CATIA; SILVA, ISABEL; CORREIA-DE-SA, PAULO; CASTELUCCI, PATRICIA. The Effect of Ischemia and Reperfusion on Enteric Glial Cells and Contractile Activity in the Ileum. Digestive Diseases and Sciences, v. 60, n. 9, p. 2677-2689, . (08/08728-5, 12/06434-0, 08/05314-5, 12/00259-1)

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