Nitric Oxide Regulates the Levels of cGMP Accumulation in the Cricket Brain


Cricket brains were incubated in a saline containing nitric oxide (NO)-donor and phosphodiesterase inhibitor IBMX, which could activate soluble guanylate cyclase (sGC) to increase cGMP levels in the targets of NO. The increase of cGMP was detected by immunohistochemistry and enzyme linked immunosorbent assay. NO-induced cGMP immunohistochemistry revealed that many cell bodies of cricket brain showed cGMP immunoreactivity when preparations were treated with a saline containing 10 mM NO-donor SNP and phosphodiesterase inhibitor IBMX, but only a few cell bodies showed immunoreactivity when preparations were incubated without NO-donor. The concentration of cGMP in cricket brains were then measured by using cGMP-specific enzyme linked immunosorbent assay. Cricket brains were treated with a saline containing 1 μM of NO-donor NOR3 and 1 mM IBMX. The cGMP levels in the brain were increased about 75% compared to control preparations that was treated with a cricket saline containing IBMX. The level of cGMP decreased about 40% when preparations were incubated NOR3 saline containing sGC inhibitor ODQ. These results indicate that NO activates sGC and increases the levels of cGMP in particular neurons of the cricket brain and that the level of cGMP would be kept a particular level, which might regulate synaptic efficacy in the neurotransmission.


  1. 1.

    Schuman, E. M., Madison, D. V. (1994) Nitric oxide and synaptic function. Annu. Rev. Neurosci. 17, 153–183.

    CAS  Article  Google Scholar 

  2. 2.

    Fujie, S., Aonuma, H., Ito, I., Gelperin, A., Ito, E. (2002) The nitric oxide/cyclic GMP pathway in the olfactory processing system of the terrestrial slug Limax marginatus. Zool. Sci. 19, 15–26.

    CAS  Article  Google Scholar 

  3. 3.

    Gelperin, A. (1994) Nitric oxide mediates network oscillations of olfactory interneurons in a terrestrial mollusc. Nature 369, 61–63.

    CAS  Article  Google Scholar 

  4. 4.

    Aonuma, H., Newland, P. L. (2001) Opposing actions of nitric oxide on synaptic inputs of identified interneurones in the central nervous system of the crayfish. J. Exp. Biol. 204, 1319–1332.

    CAS  PubMed  Google Scholar 

  5. 5.

    Aonuma, H., Newland, P. L. (2002) Synaptic inputs onto spiking local interneurons in crayfish are depressed by nitric oxide. J. Neurobiol. 52, 144–155.

    CAS  Article  Google Scholar 

  6. 6.

    Aonuma, H., Nagayama, T., Takahata, M. (2000) Modulatory effects of nitric oxide on synaptic depression in the crayfish neuromuscular system. J. Exp. Biol. 203, 3595–3602.

    CAS  PubMed  Google Scholar 

  7. 7.

    Moncada, S., Palmer, R. M. J., Higgs, E. A. (1991) Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol. Rev. 43, 109–142.

    CAS  PubMed  Google Scholar 

  8. 8.

    Bredt, D. S., Synder, S. H. (1989) Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum. Proc. Natl. Acad. Sci. USA 86, 9030–9033.

    CAS  Article  Google Scholar 

  9. 9.

    de Vente, J., Steinbusch, H. W., Schipper, J. (1987) A new approach to immunocytochemistry of 3’,5’-cyclic guanosine monophosphate: preparation, specificity, and initial application of a new antiserum against formaldehyde-fixed 3’,5’-cyclic guanosine monophosphate. Neuroscience 22, 361–373.

    Article  Google Scholar 

  10. 10.

    Ott, S. R., Jones, I. W., Burrows, M., Elphick, M. R. (2000) Sensory afferents and motor neurons as targets for nitric oxide in the locust. J. Comp. Neurol. 422, 521–532.

    CAS  Article  Google Scholar 

  11. 11.

    Aonuma, H. (2002) Distribution of NO-induced cGMP-like immunoreactive neurones in the abdominal nervous system of the crayfish, Procambarus clarkii. Zool. Sci. 19, 969–979.

    Article  Google Scholar 

  12. 12.

    Müller, U. (1997) The nitric oxide system in insects. Prog. Neurobiol. 51, 363–381.

    Article  Google Scholar 

  13. 13.

    Philippides, A., Husbands, P., O’Shea, M. (2000) Four-dimensional neuronal signaling by nitric oxide: a computational analysis. J. Neurosci. 20, 1199–1207.

    CAS  Article  Google Scholar 

  14. 14.

    East, S. J., Garthwaite, J. (1991) NMDA receptor activation in rat hippocampus induces cyclic GMP formation through the L-arginine-nitric oxide pathway. Neurosci. Lett. 123, 17–19.

    CAS  Article  Google Scholar 

  15. 15.

    Bredt, D. S., Snyder, S. H. (1992) Nitric oxide, a novel neuronal messenger. Neuron 8, 3–11.

    CAS  Article  Google Scholar 

  16. 16.

    Müller, U. (1996) Inhibition of nitric oxide synthase impairs a distinct form of long-term memory in the honeybee, Apis mellifera. Neuron 16, 541–549.

  17. 17.

    Aonuma, H., Nagao, T., Nagayama, T., Takahata, M. (1999) Modulatory effects of amino acids upon the neuromuscular transmission in the crayfish fast flexor muscle. J. Exp. Zool. 283, 531–540.

    CAS  Article  Google Scholar 

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Correspondence to H. Aonuma.

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Presented at the 10th ISIN Symposium on Invertebrate Neurobiology, July 5–9, 2003, Tihany, Hungary.

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Aonuma, H., Niwa, K. Nitric Oxide Regulates the Levels of cGMP Accumulation in the Cricket Brain. BIOLOGIA FUTURA 55, 65–70 (2004).

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  • Insect brain
  • cricket
  • soluble guanylate cyclase
  • nitric oxide
  • cGMP