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Antibodies to neurofilament protein and other brain proteins reveal the innervation of peripheral organs

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Summary

Monoclonal and polyclonal antibodies to neurofilament proteins, neuron-specific enolase, glial fibrillary acidic protein and S-100 have been used to demonstrate nerves, ganglion cells and the supportive glial system of the innervation of various organs. The female genitalia, the urinary tract, the respiratory system, the pancreas, the heart and the skin of several mammalian species, including rat, mouse, guinea pig, cat, pig, monkey and man were fixed in parabenzoquinone and portions of each organ were snap frozen. Serial or free-floating thick cryostat sections were stained using indirect immunofluorescence and peroxidase anti-peroxidase immunocytochemistry. In addition, the newly described and highly sensitive immunogold-silver staining technique was used on Bouin's-fixed and wax-embedded tissues.

Antibodies to neurofilament proteins seemed to react with neuronal structures in all the species studied. Alternately stained serial sections showed a similar distribution of neurofilament proteins and neuron-specific enolase-containing nerves. Neuron-specific enolase staining had a diffuse appearance and was found to be highly variable, indicating that the neuron-specific enolase content might be related to the physiological state of the nerves and ganglion cells, whereas antibodies to neurofilament protein gave a consistently intense and very clear picture of the ganglion cells and nerve fibres. Antibodies to S-100 stained supportive elements of the peripheral nervous system in all tissues examined, whereas antibodies to glial fibrillary acidic protein were more selective.

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Abbreviations

GFAP:

glial fibrillary acidic protein

NSE:

neuron-specific enolase

PBS:

phosphate-buffered saline

PAP:

peroxidase anti-peroxidase

FITC:

fluorescein-isothiocyanate

References

  • Bishop AE, Polak JM, Bloom SR, Pearse AGE (1978) A new universal technique for the immunocytochemical localisation of peptidergic innervation. J Endocrinol 77:25–26

    Google Scholar 

  • Bishop AE, Polak JM, Facer P, Ferri GL, Marangos PJ, Pearse AGE (1982) Neuron specific enolase: a common marker for endocrine cells and innervation of the gut and pancreas. Gastroenterology 83:902–912

    Google Scholar 

  • Bishop AE, Carlei F, Lee V, Trojanowski J, Marangos P, Dahl D, Polak JM (1985) Combined immunostaining of neurofilaments, neuron specific enolase, GFAP and S-100. A possible means for assessing the morphological and functional status of the enteric nervous system. Histochemistry 82:93–97

    Google Scholar 

  • Björklund H, Dahl D, Seiger A (1984) Neurofilament and glial fibrillary acidic protein-related immunoreactivity in rodent enteric nervous system. Neuroscience 12:277–287

    Google Scholar 

  • Bock E, Dissing J (1975) Demonstration of enolase activity connected to the brain specific protein 14-3-2. Scand J Immunol (Suppl 2) 4:31–36

    Google Scholar 

  • Brandtzaeg P (1981) Prolonged incubation staining of immunoglobulins and epithelial components in ethanol- and formaldehyde-fixed paraffin-embedded tissues. J Histochem Cytochem 29:1302–1315

    Google Scholar 

  • Coons AH, Leduc SEH, Connolly J (1955) Studies on antibody production. J Exp Med 102:49–60

    Google Scholar 

  • Dahl D, Bignami A (1983) The glial fibrillary acidic (GFA) protein and astrocytic 10-nanometer filaments. In: Lajtha A (ed), Handbook of neurochemistry, vol. 5. Plenum Press, New York, pp 78–93

    Google Scholar 

  • Dahl D, Chi NH, Miles LE, Nguyen BT, Bignami A (1982) Glial fibrillary acidic (GF) protein in Schwann cells: Fact or artefact? J Histochem Cytochem 30:912–918

    Google Scholar 

  • Dahl D, Grossi M, Bignami A (1984) Masking of epitopes in tissue sections. A study of glial fibrillary acidic (GFA) protein with antisera and monoclonal antibodies. Histochemistry 81:525–531

    Google Scholar 

  • De Vries GH, Norton WT, Raine CS (1972) Axons: Isolation from mammalian central nervous system. Science 175:1370–1372

    Google Scholar 

  • Ferri GL, Probert L, Coccia D, Michetti F, Marangos PL, Polak JM (1982) Evidence for the presence of S-100 protein in the glial component of the human enteric nervous system. Nature 297:409–410

    Google Scholar 

  • Goldman JE, Schaumburg HH, Norton WT (1978) Isolation and characterisation of glial filaments from human brain. J Cell Biol 18:426–440

    Google Scholar 

  • Goldstein ME, Sternberger LA, Sternberger NH (1983) Microheterogeneity (“neurotypy”) of neurofilament proteins. Proc Natl Acad Sci USA 80:3003–3005

    Google Scholar 

  • Graham DI, Thomas DTG, Brown I (1983) Nervous system antigens. Histopathology 7:1–21

    Google Scholar 

  • Gu J, Polak JM, Tapia FJ, Marangos PJ, Pearse AGE (1981) Neuron-specific enolase in the Merkel cells of mammalian skin. The use of specific antibody as a simple and reliable histologic marker. Am J Pathol 104:63–68

    Google Scholar 

  • Gu J, Polak JM, Van Noorden S, Pearse AGE, Marangos PJ, Azzopardi J (1983) Immunostaining of neuron specific enolase as a diagnostic tool for Merkel cell tumours. Cancer 52:1039–1043

    Google Scholar 

  • Hacker GW, Springall DR, Van Noorden S, Bishop AE, Grimelius L, Polak JM (1985) The immunogold-silver staining method — a powerful tool in histopathology. Virchows Archiv in press

  • Hickley WF, Lee V, Trojanowski JO, McMillan LJ, McKearn TJ, Gonatas J, Gonatas NK (1983) Immunohistochemical application of monoclonal antibodies against myelin basic protein and neurofilament triple protein subunits: advantages over antisera and technical limitations. J Histochem Cytochem 31:1126–1135

    Google Scholar 

  • Hoffman PN, Lasek RJ (1975) The slow axonal transport. Identification of major structural polypeptides of the axon and their generality among mammalian neurons. J Cell Biol 66:351–366

    Google Scholar 

  • Holgate CS, Jackson P, Cowen PN, Bird CC (1983) Immunogold-silver staining: a new method of immunostaining with enhanced sensitivity. J Histochem Cytochem 31:938–994

    Google Scholar 

  • Huan WM, Gibson SJ, Facer P, Gu J, Polak JM (1983) Improved section adhesion for immunocytochemistry using high molecular weight polymers of L-lysine as a slide coating. Histochemistry 77:275–297

    Google Scholar 

  • Jessen KR, Mirsky R (1980) Glial cells in the enteric nervous system contain glial fibrillary acidic protein. Nature 286:736–737

    Google Scholar 

  • Lazarides E (1980) Intermediate filaments as mechanical integrators of cellular space. Nature 283:249–256

    Google Scholar 

  • Marangos PJ, Schmechel DE (1980) The neurobiology of the brain enolases. In: Youdim MBH, Lowenberg W, Sharman DF, Lagnado JR (eds), Essays in neurochemistry and neuropharmacology, vol. 4. John Wiley & Sons, New York, pp 212–247

    Google Scholar 

  • Marangos PJ, Zomzely-Neurath C, York C (1975) Immunological studies of a nerve specific protein. Arch Biochem Biophys 170:289–293

    Google Scholar 

  • Marangos PJ, Schmechel DE, Oertel WH (1981) Neuron specific enolase: a functional marker for the diffuse neuroendocrine system. In: Bloom SR, Polak JM (eds) Gut hormones, 2nd ed. Churchill Livingstone, Edinburgh, pp 101–106

    Google Scholar 

  • Moore BW (1972) Chemistry and biology of two proteins, S-100 and 14-3-2, specific to the nervous system. International Review of Neurobiology, vol 15. Academic Press, New York, pp 215–225

    Google Scholar 

  • Ramaekers FCS, Puts JJG, Moesker O, Kant A, Huysmans A, Haag D, Jap PHK, Herman CJ, Vooijs GP (1983) Antibodies to intermediate filament proteins in the immunohistochemical identification of human tumours: an overview. Histochem J 15:691–713

    Google Scholar 

  • Rodrigo, J, Polak JM, Terenghi G, Cervantes C, Ghatei MA, Mulderry PK, Bloom SR (1985) Calcitonin gene-related peptide (CGRP)-immunoreactive sensory and motor nerves of the mammalian palate. Histochemistry 82:67–74

    Google Scholar 

  • Schmechel D, Marangos PJ, Brightman M (1978) Neuron specific enolase is a molecular marker for the peripheral and central neuroendocrine cells. Nature 276:834–836

    Google Scholar 

  • Shaw G, Weber K (1981) The distribution of the neurofilament triplet proteins within individual neurones. Exp Cell Res 136:119–125

    Google Scholar 

  • Sheppard MN, Kurian SS, Henzen-Logmans SC, Michetti F, Cocchia D, Cole P, Rush RA, Marangos PJ, Bloom SR, Polak JM (1983) Neuron specific enolase and S-100: new markers for delineating the innervation of the respiratory tract in man and other mammals. Thorax 38:333–340

    Google Scholar 

  • Sheppard MN, Marangos PJ, Polak JM, Bloom SR (1984) Neuron specific enolase: a marker for the early development of nerves and endocrine cells in the human lung. Life Sci 34:265–271

    Google Scholar 

  • Springall DR, Hacker GW, Grimelius L, Polak JM (1984) The potential of the immunogold-silver staining method for paraffin sections. Histochemistry 81:603–608

    Google Scholar 

  • Sternberger LA (1979) The unlabelled antibody peroxidase antiperoxidase (PAP) method. In: Sternberger LA (ed) Immunocytochemistry, 2nd edn. John Wiley & Sons, New York, pp 104–169

    Google Scholar 

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Authors and Affiliations

  1. Department of Histochemistry, Hammersmith Hospital, Royal Postgraduate Medical School, Du Cane Road, W12 OHS, London, UK

    G. W. Hacker, J. M. Polak, D. R. Springall, J. Ballesta, A. Cadieux & J. Gu

  2. Division of Neuropathology, Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, USA

    J. Q. Trojanowski

  3. Department of Neuropathology, Harvard Medical School, and Spinal Cord Injury Research Laboratory, West Roxbury Veterans Administration Medical Centre, Boston, Massachussetts, USA

    D. Dahl

  4. Clinical Psychobiology Branch, National Institute of Mental Health, Bethesda, Maryland, USA

    P. J. Marangos

Authors
  1. G. W. Hacker
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  2. J. M. Polak
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  3. D. R. Springall
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  4. J. Ballesta
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  5. A. Cadieux
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  6. J. Gu
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  7. J. Q. Trojanowski
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  8. D. Dahl
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  9. P. J. Marangos
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Hacker, G.W., Polak, J.M., Springall, D.R. et al. Antibodies to neurofilament protein and other brain proteins reveal the innervation of peripheral organs. Histochemistry 82, 581–593 (1985). https://doi.org/10.1007/BF00489980

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  • DOI: https://doi.org/10.1007/BF00489980

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