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Effects of Ischemia and Reperfusion on Subpopulations of Rat Enteric Neurons Expressing the P2X7 Receptor

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Abstract

Background

Intestinal ischemia followed by reperfusion (I/R) may occur following intestinal obstruction. In rats, I/R in the small intestine leads to structural changes accompanied by neuronal death.

Aim

To analyze the impact of I/R injury on different neuronal populations in the myenteric plexus of rat ileum.

Methods

The ileal artery was occluded for 35 min and animals were euthanized 6, 24, and 72 h, and 1 week later. Immunohistochemistry was performed with antibodies against the P2X7 receptor as well as nitric oxide synthase (NOS), calbindin, calretinin, choline acetyltransferase (ChAT), or the pan-neuronal marker anti-HuC/D.

Results

Double immunolabeling demonstrated that 100 % of NOS-, calbindin-, calretinin-, and ChAT-immunoreactive neurons in all groups expressed the P2X7 receptor. Following I/R, neuronal density decreased by 22.6 % in P2X7 receptor-immunoreactive neurons, and decreased by 46.7, 38, 39.8, 21.7, and 20 % in NOS-, calbindin-, calretinin-, ChAT-, and HuC/D-immunoreactive neurons, respectively, at 6, 24, and 72 h and 1 week following injury compared to the control and sham groups. We also observed a 14 % increase in the neuronal cell body profile area of the NOS-immunoreactive neurons at 6 and 24 h post-I/R and a 14 % increase in ChAT-immunoreactive neurons at 1 week following I/R. However, the average size of the calretinin-immunoreactive neurons was reduced by 12 % at 6 h post-I/R and increased by 8 % at 24 h post-I/R.

Conclusions

This work demonstrates that I/R is associated with a significant loss of different subpopulations of neurons in the myenteric plexus accompanied by morphological changes, all of which may underlie conditions related to intestinal motility disorder.

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Acknowledgments

We would like to thank Professors Jackson Cioni Bittencourt and Carol Fuzeti Elias for use of the fluorescence microscope for some analyses and Rosana Prisco for statistical analysis. These studies were supported by FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo Proc 08/05314-5) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior).

Conflict of interest

None.

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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, São Paulo, CEP 05508-900, Brazil

    Kelly Palombit, Cristina Eusébio Mendes, Mariana Póvoa Silveira & Patricia Castelucci

  2. Department of Pharmacology, Biomedical Science Institute, University of São Paulo, São Paulo, Brazil

    Wothan Tavares-de-Lima

  3. Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil

    Mariana Póvoa Silveira

Authors
  1. Kelly Palombit
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  2. Cristina Eusébio Mendes
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  4. Mariana Póvoa Silveira
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  5. Patricia Castelucci
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Correspondence to Patricia Castelucci.

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Palombit, K., Mendes, C.E., Tavares-de-Lima, W. et al. Effects of Ischemia and Reperfusion on Subpopulations of Rat Enteric Neurons Expressing the P2X7 Receptor. Dig Dis Sci 58, 3429–3439 (2013). https://doi.org/10.1007/s10620-013-2847-y

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