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Effects of Ischemia and Reperfusion on P2X2 Receptor Expressing Neurons of the Rat Ileum Enteric Nervous System

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Abstract

Purpose

We investigated the effects of ischemia/reperfusion in the intestine (I/R-i) on purine receptor P2X2-immunoreactive (IR) neurons of the rat ileum.

Methods

The superior mesenteric artery was occluded for 45 min with an atraumatic vascular clamp and animals were sacrificed 4 h later. Neurons of the myenteric and submucosal plexuses were evaluated for immunoreactivity against the P2X2 receptor, nitric oxide synthase (NOS), choline acetyl transferase (ChAT), calbindin, and calretinin.

Results

Following I/R-i, we observed a decrease in P2X2 receptor immunoreactivity in the cytoplasm and surface membranes of neurons of the myenteric and submucosal plexuses. These studies also revealed an absence of calbindin-positive neurons in the I/R-i group. In addition, the colocalization of the P2X2 receptor with NOS, ChAT, and calretinin immunoreactivity in the myenteric plexus was decreased following I/R-i. Likewise, the colocalization between P2X2 and calretinin in neurons of the submucosal plexus was also reduced. In the I/R-i group, there was a 55.8% decrease in the density of neurons immunoreactive (IR) for the P2X2 receptor, a 26.4% reduction in NOS-IR neuron, a 25% reduction in ChAT-IR neuron, and a 47% reduction in calretinin-IR neuron. The density of P2X2 receptor and calretinin-IR neurons also decreased in the submucosal plexus of the I/R-i group. In the myenteric plexus, P2X2-IR, NOS-IR, ChAT-IR and calretinin-IR neurons were reduced in size by 50%, 49.7%, 42%, and 33%, respectively, in the I/R-i group; in the submucosal plexus, P2X2-IR and calretinin-IR neurons were reduced in size by 56% and 72.6%, respectively.

Conclusions

These data demonstrate that ischemia/reperfusion of the intestine affects the expression of the P2X2 receptor in neurons of the myenteric and submucosal plexus, as well as density and size of neurons in this population. Our findings indicate that I/R-i induces changes in P2X2-IR enteric neurons that could result in alterations in intestinal motility.

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Acknowledgments

We would like to thank Professor Edson Aparecido Liberti for facilities, Professor Jackson Cioni Bittencourt and Assistant Professor Carol Fuzeti Elias for use of the Leica Microscope in these analyses. This work was supported by the FAPESP (Fundação de Amparo Pesquisa do Estado de São Paulo 05/04752-0) and CNPq.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Anatomy, Institute of Biomedical Sciences, University of São Paulo, Av. Prof. Dr. Lineu Prestes, 2415, CEP 05508-900, São Paulo, Brazil

    Ariane Silva Paulino, Kelly Palombit, Márcia Sanae Mizuno, Aline Rosa Marosti, Marcos Vinícius da Silva, Priscila Azevedo Girotti, Edson Aparecido Liberti & Patricia Castelucci

  2. Department of Pharmacology, Institute of Biomedical Sciences, University of São Paulo, Av. Prof. Dr. Lineu Prestes, 2415, CEP 05508-900, São Paulo, Brazil

    Gabriela Cavriani & Wothan Tavares-de-Lima

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  1. Ariane Silva Paulino
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  2. Kelly Palombit
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  3. Gabriela Cavriani
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Correspondence to Patricia Castelucci.

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Paulino, A.S., Palombit, K., Cavriani, G. et al. Effects of Ischemia and Reperfusion on P2X2 Receptor Expressing Neurons of the Rat Ileum Enteric Nervous System. Dig Dis Sci 56, 2262–2275 (2011). https://doi.org/10.1007/s10620-011-1588-z

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