Skip to main content
SpringerLink
Log in

Heme Oxygenase-2 and Nitric Oxide Synthase Immunoreactivity of Bovine Olfactory Receptor Neurons and a Comparison with the Distribution of NADPH-diaphorase Staining

  • Published:
The Histochemical Journal Aims and scope Submit manuscript

    We’re sorry, something doesn't seem to be working properly.

    Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Abstract

It has recently been suggested that, in addition to nitric oxide (NO), carbon monoxide (CO) is an important gaseous messenger which might be involved in vertebrate olfactory transduction because its effects include activation of guanylyl cyclase and the formation of cGMP. As there is no information regarding the presence of heme oxygenase-2 – the constitutive isoform of the heme oxygenase system – in olfactory neurons of non-rodent species, we have investigated the distribution pattern of heme oxygenase-2 in the olfactory epithelium of the bovine, a representative of macrosmatics. Localization of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity of the olfactory epithelium was compared with heme oxygenase-2 and NO synthase (NOS) immunoreactivities in order to obtain possible hints at functional significance. NADPH-d activity was particularly intense in apical dendrites of receptor neurons. It was also found in Bowman glands and intraepithelial duct cells. Less intense, discrete NADPH-d activity was present also at intermediate and basal levels of the olfactory epithelium, corresponding to the layer of receptor neuron somata and basal cells. While heme oxygenase-2 activity mainly occured in neuronal perikarya, a very intense NOS immunoreactivity, exclusively for the inducible isoform, was detected in the apical dendrites. Ultrastructurally, NADPH-d histochemistry showed distinct labelling of membranes, in particular of endoplasmic reticulum, mitochondria and nucleus. The coincident localization of the moderate NADPH-d activity and heme oxygenase-2 immunoreactivity in receptor cell perikarya suggest a functional association between NADPH-cytochrome P450 reductase and heme oxygenase-2. In contrast, dendritic localization of NADPH-d activity is topically and possibly functionally related to the presence of the inducible isoform of NOS. The results suggest that both CO and NO may be generated in bovine receptor neurons and thus involved in odorant stimulation. Based on immunocytochemical localization of synthesizing enzymes, NO might be regarded as a direct regulator of transduction related processes while CO might act as a modulator of the initial signal.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References cited

  • Arnhold S, Andressen C, Bloch W, Mai JK, Addicks K (1997) NO synthase II is transiently expressed in embyonic mouse olfactory neurons. Neurosci Lett 229: 165-169.

    Google Scholar 

  • Bredt DS, Snyder SH (1992) Nitric oxide, a novel neuronal messenger. Neuron 8: 3-11.

    Google Scholar 

  • Bredt DS, Hwang PM, Glatt CE, Lowenstein C, Reed RR, Snyder SH (1991) Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase. Nature 351: 714-718.

    Google Scholar 

  • Breer H, Shepherd GM (1993) Implications of the NO/c-GMP system for olfaction. Trends Neurosci 16: 5-9.

    Google Scholar 

  • Breer H, Klemm T, Boeckhoff I (1992) Nitric oxide mediated formation of cyclic GMP. Neuroreport 3: 1030-1032.

    Google Scholar 

  • Broillet MC, Firestein S (1996) Direct activation of the olfactory cyclic nucleotide-gated channel through modification of sulfhydryl groups by NO compounds. Neuron 16: 377-385.

    Google Scholar 

  • Broillet MC, Firestein S (1997) β-Subunits of the olfactory cyclic nucleotide-gated channel forms nitric oxide activated Ca2+ channel. Neuron 18: 951-958.

    Google Scholar 

  • Calka J, Wolf G, Schmidt W (1996) Induction of cytosolic NADPHdiaphorase/ nitric oxide synthase in reactive microglia/macrophages after quinolinic acid lesions in rat striatum: an electron and light microscopical study. Histochem Cell Biol 105: 81-89.

    Google Scholar 

  • Cattel V, Jansen A (1995) Inducible nitric oxide synthase in inflammation. Histochem J 27: 777-784.

    Google Scholar 

  • Dellacorte C, Kalinoski DL, Huque T, Wysocki L, Restrepo D (1995) NADPH diaphorase staining suggests localization of nitric oxide synthase within mature vertebrate olfactory neurons. Neuroscience 66: 215-225.

    Google Scholar 

  • Elmer HL, Brady KG, Drumm ML, Kelley TJ (1999) Nitric oxide-mediated regulation of transepithelial sodium and chloride transport in murine nasal epithelium. Am J Physiol 276: 466-473.

    Google Scholar 

  • Ewing JF, Weber CM, Maines MD (1993) Biliverdin reductase is heat resistant and coexpressed with constitutive and heat shock forms of heme oxygenase in brain. J Neurochem 61: 1015-1023.

    Google Scholar 

  • Furukawa K, Harrison DG, Saleh D, Shennib H, Chagnon FP, Giaid A (1996) Expression of nitric oxide synthase in the human nasal mucosa. Am J Respir Crit Care Med 153: 847-850.

    Google Scholar 

  • Galea E, Reis DJ, Xu H, Feinstein DL (1995) Transient expression of calcium-independent nitric oxide synthase in blood vessels during brain development. FASEB J 9: 1632-1637.

    Google Scholar 

  • Gonzales-Hernandez T, Perez De La Cruz MA, Mantolan-Sarmiento B (1996) Histochemical and immunohistochemical detection of neurons that produce nitric oxide: effect of different fixative parameters and immunoreactivity against non-neuronal NOS antisera. J Histochem Cytochem 44: 1399-1413.

    Google Scholar 

  • Guo FH, Deraeve HR, Rice TW, Stuehr DJ, Thunnisen FBJM, Erzurum SC (1995) Continuous nitric oxide synthesis by inducible nitric oxide synthase in normal human airway epithelium in vivo. Proc Natl Acad Sci USA 92: 7809-7813.

    Google Scholar 

  • Hope BT, Michael GJ, Knigge KM, Vincent RS (1991) Neuronal NADPH diaphorase is a nitric oxide synthase. Proc Natl Acad Sci USA 88: 2811-2814.

    Google Scholar 

  • Ingi T, Ronnett GV (1995) Direct demonstration of a physiological role of carbon monoxide in olfactory receptor neurons. J Neurosci 15: 8214-8222.

    Google Scholar 

  • Ingi T, Chiang G, Ronnett GV (1996) The regulation of heme turnover and carbon monoxide biosynthesis in cultured primary rat olfactory receptor neurons. J Neurosci 16: 5621-5628.

    Google Scholar 

  • Jacob JM, Dorheim MA, Grammas P (1999) The effect of age and injury on the expression of inducible nitric oxide synthase in facial motor neurons in F344 rats. Mech Ageing Dev 107: 205-218.

    Google Scholar 

  • Kelley TJ, DrummML (1998) Inducible nitric oxide synthase expression is reduced in cystic fibrosis murine and human airway epithelial cells. J Clin Invest 102: 1200-1207.

    Google Scholar 

  • Kishimoto J, Keverne EB, Hardwick J, Emson PC (1993) Localization of nitric oxide synthase in the mouse olfactory and vomeronasal system: a histochemical, immunological and in situ hybridization study. Eur J Neurosci 5: 1684-1694.

    Google Scholar 

  • Kulkarni AP, Getchell TV, Getchell ML (1994) Neuronal nitric oxide synthase is localized in extrinsic nerves regulating perireceptor processes in the chemosensory nasal mucosa of rats and humans. J Comp Neurol 345: 125-138.

    Google Scholar 

  • Leinders-Zufall T, Shepherd GM, Zufall F (1995) Regulation of cyclic nucleotide-gated channels and membrane excitability in olfactory receptor cells by carbon monoxide. J Neurophysiol 74: 1498-1508.

    Google Scholar 

  • Leinders-Zufall T, Shepherd GM, Zufall F (1996) Modulation of cyclic GMP of the odour sensitivity of vertebrate olfactory receptor cells. Proc R Soc Lond B Biol Sci 263: 803-811.

    Google Scholar 

  • Maines MD (1997) The heme oxygenase system: a regulator of second messenger gases. Annu Rev Pharmacol Toxicol 37: 517-554.

    Google Scholar 

  • Marletta MA (1994) Nitric oxide synthase: aspects concerning structure and catalysis. Cell 78: 927-930.

    Google Scholar 

  • Menco BPM (1997) Ultrastructural aspects of olfactory signaling. Chem Senses 22: 295-311.

    Google Scholar 

  • Morales B, Bacigalupo J (1996) Chemical reception in vertebrate olfaction: evidence for multiple pathways. Biol Res 29: 333-341.

    Google Scholar 

  • Nakamura T, Gold GH (1987) A cyclic-nucleotide gated conductance in olfactory receptor cilia. Nature 325: 442-444.

    Google Scholar 

  • Nathan C, Xie QW (1994) Nitric oxide synthases: roles, tolls, and controls. Cell 78: 915-918.

    Google Scholar 

  • Norris PJ, Hardwick JP, Emson PC (1994) Localization of NADPH cytochrome P450 oxidoreductase in rat brain by immunohistochemistry and in situ hybridization and a comparison with the distribution of neuronal NADPH-diaphorase staining. Neuroscience 61: 331-350.

    Google Scholar 

  • Norris PJ, Charles IG, Scorer CA, Emson PC (1995) Studies on the localization and expression of nitric oxide synthase using histochemical techniques. Histochem J 27: 745-756.

    Google Scholar 

  • Ramos KS, Lin H, Mcgrath JJ (1989) Modulation of cyclic guanosine monophosphate levels in cultured aortic smooth muscle cells by carbon monoxide. Biochem Pharmacol 38: 1368-1370.

    Google Scholar 

  • Rosbe KW, Mims JW, Prazma J, Petrusz P, Rose A, Drake AF (1996) Immunohistochemical localization of nitric oxide synthase activity in upper respiratory epithelium. Laryngoscope 106: 1075-1079.

    Google Scholar 

  • Roskams AJ, Bredt DS, Dawson TM, Ronnett GV (1994) Nitric oxide mediates the formation of synaptic connections in developing and regenerating olfactory receptor neurons. Neuron 13: 289-299.

    Google Scholar 

  • Rotenberg MO, Maines MD (1991) Characterization of a cDNAencoding rabbit brain heme oxygenase-2 and identification of a conserved domain among mammalian heme oxygenase isoezymes: possible heme-binding site? Arch Biochem Biophys 290: 336-344.

    Google Scholar 

  • Rothe F, Canzler U, Wolf G (1998) Subcellular localization of the neuronal isoform of nitric oxide synthase in the brain: a critical evaluation. Neuroscience 83: 259-269.

    Google Scholar 

  • Schmachtenberg O, Bacigalupo J (1999) Nitric oxide activates a potassium current in olfactory receptor neurons from Caudiverbera and Xenopus laevis. Brain Res 837: 301-305.

    Google Scholar 

  • Shnayder-Shapiro L, Halpern M (1998) Development of NADPHdiaphorase expression in chemosensory systems of the opossum, Monodelphis domestica. Dev Brain Res 111: 51-63.

    Google Scholar 

  • Tang FR, Tan CK, Ling EA (1995) Light and electron microscopic studies of the distribution of NADPH-diaphorase in the rat upper thoracic spinal cord with special reference to the spinal autonomic region. Arch Histol Cytol 58: 493-505.

    Google Scholar 

  • Verma A, Hirsch DJ, Glatt CE, Ronnett GV, Snyder SH (1993) Carbon monoxide: a putative neural messenger. Science 259: 381-384.

    Google Scholar 

  • Vollerthun R, Hohler B, Kummer W (1995) Guinea-pig sympathetic postganglionic neurones contain heme oxygenase-2. Neuroreport 7: 173-176.

    Google Scholar 

  • Vollerthun R, Hohler B, Kummer W (1996) Heme oxygenase-2 in primary afferent neurons of the guinea-pig. Histochem Cell Biol 105: 453-458.

    Google Scholar 

  • Young HM, O'Brien AJ, Furness JB, Ciampoli D, Hardwick JP, Mccabe TJ, Narayanasami R, Masters BSS, Tracey WR (1997) Relationships between NADPH diaphorase staining and neuronal, endothelial, and inducible nitric oxide synthase and cytochrome P450 reductase immunoreactivities in guinea-pig tissues. Histochem Cell Biol 107: 19-29.

    Google Scholar 

  • Zhao H, Firestein S, Greer C (1994) NADPH-diaphorase localization in the olfactory system. Neuroreport 6: 149-152.

    Google Scholar 

  • Zielinski BS, Osahan JK, Hara TJ, Hosseini M, Wong E (1996) Nitric oxide synthase in the olfactory mucosa of the larval sea lamprey (Petromyzon marinus). J Comp Neurol 365: 18-26.

    Google Scholar 

  • Zufall F, Leinders-Zufall T (1997) Identification of a long-lasting form of odor adaption that depends on the carbon monoxide/cGMP second messenger system. J Neurosci 17: 2703-2712.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, Giessen, Frankfurter Str. 98, 35392, Giessen, Germany

    S. Wenisch & R. Leiser

  2. Department of Anatomy, University Köln, Josef-Stelzmann-Strasse 9, D-50931, Köln, Germany

    C. Andressen, S. Arnhold & K. Addicks

  3. Institute of Anatomy, University Dresden, Fetscherstrasse 74, D-01307, Dresden, Germany

    A. Derouiche

Authors
  1. S. Wenisch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. Andressen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Derouiche
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S. Arnhold
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K. Addicks
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. R. Leiser
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wenisch, S., Andressen, C., Derouiche, A. et al. Heme Oxygenase-2 and Nitric Oxide Synthase Immunoreactivity of Bovine Olfactory Receptor Neurons and a Comparison with the Distribution of NADPH-diaphorase Staining. Histochem J 32, 381–388 (2000). https://doi.org/10.1023/A:1004021832449

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1004021832449

Keywords

Search

Navigation