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Regional distribution of binding sites for the nitric oxide synthase inhibitorl-[3H]Nitroarginine in rat brain

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Abstract

The regional distribution of NG-nitro-l-[3H]arginine (L-[3H]NOARG) binding to different regions of rat brain was studied by quantitative autoradiography. These studies revealed highest density of binding sites in cerebellum, anterior olfactory nucleus, islands of Calleja and substantia nigra with appreciable binding site densities in inferior colliculus, superior colliculus, olfactory tubercle and dorsal tegmental nucleus. The regional distribution of L-[3H]NOARG binding, is in good agreement with the distribution of nitric oxide synthase studied previously by NADPH-diaphorase staining and immunohistochemistry using antibodies against neuronal nitric oxide synthase. The kinetics of L-[3H]NOARG binding to the cytosolic preparations of cerebral cortex, cerebellum, hippocampus and striatum was studied using an in vitro binding technique. Specific L-[3H]NOARG binding was of nanomolar affinity, saturable, and best fit to a single-site model in all four brain regions. These studies support the potential use of L-[3H]NOARG binding as a tool for further elucidation of the regional distribution and functional properties of NOS in the central nervous system.

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References

  1. Dawson, T. M., and Snyder, S. H. 1994. Gases as biological messengers: Nitric oxide and carbon monoxide in the brain. J. Neurosci. 14:5147–5159.

    PubMed  CAS  Google Scholar 

  2. Garthwaite, J. 1995. Neural nitric oxide signalling. Trends Neurosci. 18:51–52.

    Article  PubMed  CAS  Google Scholar 

  3. Hecker, M., Mülsch, A., and Busse, R. 1994. Subcellular localization and characterization of neuronal nitric oxide synthase. J. Neurochem. 62:1524–1529.

    Article  PubMed  CAS  Google Scholar 

  4. Murphy, S., Simmons, M. L., Agullo, L., Garcia, A., Feinstein, D. L., Galea, E., Reis, D. J., Minc-Golomb, D., and Schwartz, J. P. 1993. Synthesis of nitric oxide in CNS glial cells. Trends Neurosci. 16:323–328.

    Article  PubMed  CAS  Google Scholar 

  5. Thomas, E., and Pearse, A. G. E. 1961. The fine localization of dehydrogenases in the nervous system. Histochemie 2:262–282.

    Article  Google Scholar 

  6. Dawson, T. M., Bredt, D. S., Fotuhi, M., Hwang, P. M., and Snyder, S. H. 1991. Nitric oxide synthase and neuronal NADPH-diaphorase are identical in brain and peripheral tissues. Proc. Natl. Acad. Sci. USA 88:7797–7801.

    Article  PubMed  CAS  Google Scholar 

  7. Vincent, S. R., and Kimura, H. 1992. Histochemical mapping of nitric oxide synthase in the rat brain. Neuroscience 46:755–784.

    Article  PubMed  CAS  Google Scholar 

  8. Tracey, W. R., Nakane, M., Pollock, J. S., and Förstermann, U. 1993. Nitric oxide synthases in neuronal cells, macrophages and endothelium are NAPDH diaphorases, but represents only a fraction of total cellular NADPH diaphorase activity. Biochem. Biophys. Res. Commun. 195:1035–1040.

    Article  PubMed  CAS  Google Scholar 

  9. Matsumoto, T., Nakane, M., Pollock, J. S., Kuk, J. E., and Förstermann, U. 1993. A correlation between soluble brain nitric oxide synthase and NADPH-diaphorase activity is only seen after exposure of the tissue to fixative. Neurosci. Lett. 155:61–64.

    Article  PubMed  CAS  Google Scholar 

  10. Bredt, D. S., Hwang, P. M., and Snyder, S. H. 1990. Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature 347:768–770.

    Article  PubMed  CAS  Google Scholar 

  11. Kishimoto, J., Keverne, E. B., Hardwick, J., and Emson, P. C. 1993. Localization of nitric oxide synthase in the mouse olfactory and vomeronasal system: a histochemical, immunological andin situ hybridization study. Eur. J. Neurosci. 5:1684–1694.

    Article  PubMed  CAS  Google Scholar 

  12. Schmidt, H. H. H. W., Gagne, G. D., Nakane, M., Pollock, J. S., Miller, M. F., and Murad, F. 1992. Mapping of neural nitric oxide synthase in the rat suggests frequent co-localization with NADPH diaphorase but not with soluble guanylyl cyclase, and novel paraneural functions for nitrinergic signal transduction. J. Histochem. Cytochem. 40:1439–1456.

    PubMed  CAS  Google Scholar 

  13. Kidd, E. J., Michel, A. D., and Humphrey, P. P. 1995. Autoradiographic distribution of [3H]L-N G-nitro-arginine binding in rat brain. Neuropharmacol. 34:63–73.

    Article  CAS  Google Scholar 

  14. Rao, V. L. R., and Butterworth, R. F. 1996. Kinetics, pharmacology and autoradiographic distribution of L-[3H]nitroarginine binding sites in rat cerebellum. J. Neurochem. 66:701–709.

    PubMed  CAS  Google Scholar 

  15. Michel, A. D., Phul, R. K., Stewart, T. L., and Humphrey, P. P. 1993. Characterization of the binding of [3H]-L-NG-nitro-arginine in rat brain. Br. J. Pharmacol. 109:287–288.

    PubMed  CAS  Google Scholar 

  16. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. 1951. Protein measurements with Folin-phenol reagent. J. Biol. Chem. 193:265–275.

    PubMed  CAS  Google Scholar 

  17. Paxinos, G., and Watson, C. 1982. The Rat Brain: in Stereotaxic Coordinates. Academic Press, NY, USA.

    Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  19. Rao, V. L. R., Audet, R. M., and Butterworth, R. F. 1995. Increased nitric oxide synthase activities and L-[3H]arginine uptake in brain following portacaval anastomosis. J. Neurochem. 65:677–681.

    PubMed  CAS  Google Scholar 

  20. Brüning, G. 1993. NADPH-diaphorase histochemistry in the postnatal mouse cerebellum suggests specific developmental functions for nitric oxide. J. Neurosci. Res. 36:580–587.

    Article  PubMed  Google Scholar 

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Correspondence to R. F. Butterworth.

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Raghavendra Rao, V.L., Butterworth, R.F. Regional distribution of binding sites for the nitric oxide synthase inhibitorl-[3H]Nitroarginine in rat brain. Neurochem Res 21, 355–359 (1996). https://doi.org/10.1007/BF02531652

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