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Knock out of neuronal nitric oxide synthase exacerbates intestinal ischemia/reperfusion injury in mice

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Abstract

Recent investigation of the intestine following ischemia and reperfusion (I/R) has revealed that nitric oxide synthase (NOS) neurons are more strongly affected than other neuron types. This implies that NO originating from NOS neurons contributes to neuronal damage. However, there is also evidence of the neuroprotective effects of NO. In this study, we compared the effects of I/R on the intestines of neuronal NOS knockout (nNOS−/−) mice and wild-type mice. I/R caused histological damage to the mucosa and muscle and infiltration of neutrophils into the external muscle layers. Damage to the mucosa and muscle was more severe and greater infiltration by neutrophils occurred in the first 24 h in nNOS−/− mice. Immunohistochemistry for the contractile protein, α-smooth muscle actin, was used to evaluate muscle damage. Smooth muscle actin occurred in the majority of smooth muscle cells in the external musculature of normal mice but was absent from most cells and was reduced in the cytoplasm of other cells following I/R. The loss was greater in nNOS−/− mice. Basal contractile activity of the longitudinal muscle and contractile responses to nerve stimulation or a muscarinic agonist were reduced in regions subjected to I/R and the effects were greater in nNOS−/− mice. Reductions in responsiveness also occurred in regions of operated mice not subjected to I/R. This is attributed to post-operative ileus that is not significantly affected by knockout of nNOS. The results indicate that deleterious effects are greater in regions subjected to I/R in mice lacking nNOS compared with normal mice, implying that NO produced by nNOS has protective effects that outweigh any damaging effect of this free radical produced by enteric neurons.

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Acknowledgments

We thank Associate Professor James Brock and Ms. Dorota Ferens for their helpful advice on the isometric tension recording experiments, Dr. Trung Nguyen for statistical advice and Mr. Gene Venables for his assistance in surgery and genotyping.

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Authors and Affiliations

  1. Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria, 3010, Australia

    Leni R. Rivera, Louise Pontell, Hyun-Jung Cho, Michelle Thacker, Daniel P. Poole, Tony Frugier & John B. Furness

  2. Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, 3052, Australia

    Leni R. Rivera

  3. Department of Anatomy, University of São Paulo, São Paulo, Brazil

    Patricia Castelucci

Authors
  1. Leni R. Rivera
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  2. Louise Pontell
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  3. Hyun-Jung Cho
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  4. Patricia Castelucci
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  5. Michelle Thacker
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  6. Daniel P. Poole
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  7. Tony Frugier
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  8. John B. Furness
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Correspondence to Leni R. Rivera.

Additional information

This work was supported by a grant from the National Health and Medical Research Council of Australia (grant 1006035).

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Rivera, L.R., Pontell, L., Cho, HJ. et al. Knock out of neuronal nitric oxide synthase exacerbates intestinal ischemia/reperfusion injury in mice. Cell Tissue Res 349, 565–576 (2012). https://doi.org/10.1007/s00441-012-1451-3

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  • DOI: https://doi.org/10.1007/s00441-012-1451-3

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