Skip to main content
SpringerLink
Log in

Light and electron microscopical immunocytochemistry of 5′-nucleotidase in rat cerebellum

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

5′-Nucleotidase in nervous tissue has so far not been localised at the ultrastructural level using immunocytochemical techniques. We have now applied monoclonal antibodies and a polyclonal antiserum raised against this ecto-enzyme and describe the distribution of 5′-nucleotidase antigenicity in rat cerebellum both at the light and electron microscopic levels. Within all cerebellar layers, 5′-nucleotidase immunoreactivity was found on plasma membranes of glial elements, i.e. Bergmann glial cell processes crossing the molecular layer, astrocytic end-feet around blood vessels and glial cell extensions surrounding single Purkinje cells. In the granular layer, 5′-nucleotidase immunoreactivity was present on glial membranes interposed between granule cells. Neuronal cells or processes were devoid of immunoreactivity. The immunocytochemical results were compared with conventional 5′-nucleotidase histochemistry. Both techniques showed the same ecto-localisation of the enzyme and favour the view of 5′-nucleotidase being predominantly situated at glial plasma membranes.

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

  • Amano S, Kreutzberg GW, Reddington M (1983) 5′-nucleotidase activity in human astrocytomas. Acta Neuropathol 59:145–149

    Google Scholar 

  • Andersson Forsman C, Elfvin LG (1984) Cytochemical localization of 5′-nucleotidase in guinea-pig sympathetic ganglia. J Neurocytol 13:339–349

    Google Scholar 

  • Cammer W, Sacchi R, Kahn S (1985) Immunocytochemical localization of 5′-nucleotidase in oligodendroglia and myelinated fibers in the central nervous system of adult and young rats. Dev Brain Res 20:89–69

    Google Scholar 

  • Cammer W, Tansey FA, Sacchi R (1986) Antibody against mouse liver 5′-nucleotidase immunostains white matter in the adult mouse central nervous system. J Neurol Sci 73:155–167

    Google Scholar 

  • de Lima AD, Montero VM, Singer W (1985) The cholinergic innervation of the visual thalamus: an EM immunocytochemical study. Exp Brain Res 59:206–212

    Google Scholar 

  • DePierre JW, Karnovsky ML (1974a) Ectoenzymes of the guinea pig polymorphonuclear leukocyte I. Evidence for an ecto-adenosine monophosphate.-adenosine triphosphatase, and-p-nitrophenyl phosphatase. J Biol Chem 249:7111–7120

    Google Scholar 

  • DePierre JW, Karnovsky ML (1974b) Ecto-enzyme of granulocytes: 5′-nucleotidase. Science 183:1096–1098

    Google Scholar 

  • Dieckhoff J, Mannherz HG (1985) The interaction of 5′-nucleotidase purified from chicken gizzard and actin, and the reversible loss of the inhibitory capacity of actin on deoxyribonuclease I. Biochim Biophys Acta 829:209–220

    Google Scholar 

  • Dieckhoff J, Lietzke R, Mannherz HG (1986a) Monoclonal antibodies against 5′-nucleotidase from chicken gizzard. Evidence for species and tissue specific differences of 5′-nucleotidase. J Biol Chem 261:922–927

    Google Scholar 

  • Dieckhoff J, Mollenhauer J, Kühl U, Niggemeyer B, von der Mark K, Mannherz HG (1986b) The extracellular matrix proteins laminin and fibronectin modify the AMPase activity of 5′-nucleotidase from chicken gizzard smooth muscle. FEBS Lett 195:82–86

    Google Scholar 

  • El-Aaser AA, Fitzsimons JTR, Hinton RH, Norris KA, Reid E (1973) Electron microscopic examination of hepatic subcellular fractions for phosphatases. Histochem J 5:199–223

    Google Scholar 

  • Graeber MB, Kreutzberg GW (1986) Astrocytes increase in glial fibrillary acidic protein during retrograde changes of facial motor neurons. J Neurocytol 15:363–373

    Google Scholar 

  • Freund TF, Magloczky ZS, Soltesz I, Somogyi P (1986) Synaptic connections, axonal and dendritic patterns of neurons immunoreactive for cholecystokinin in the visual cortex of the cat. Neuroscience 19:1133–1159

    Google Scholar 

  • Harb J, Meflah K, Bernard S (1985) Structural differences between plasma-membrane 5′-nucleotidase in different cell types as evidenced by antibodies. Biochem J 232:859–862

    Google Scholar 

  • Hendry SHC, Jones EG, Emson PC (1984) Morphology, distribution, and synaptic relations of somatostatin-and neuropeptide Y-immunoreactive neurons in rat and monkey neocortex. J Neurosci 4:2497–2517

    Google Scholar 

  • Holländer H (1981) Target preparation for histochemical studies with the electron microscope. Sci Tools 28:8–10

    Google Scholar 

  • Karlsson U, Schultz RL (1965) Fixation of the central nervous system for electron microscopy by aldehyde perfusion. J Ultrastruct Res 12:160–186

    Google Scholar 

  • Kreutzberg GW, Barron KD (1978) 5′-nucleotidase of microglial cells in the facial nucleus during axonal reaction. J Neurocytol 7:601–610

    Google Scholar 

  • Kreutzberg GW, Hussain ST (1982) Cytochemical heterogeneity of the glial plasma membrane: 5′-nucleotidase in retinal Müller cells. J Neurocytol 11:53–64

    Google Scholar 

  • Kreutzberg GW, Hussain ST (1984) Cytochemical localization of 5′-nucleotidase activity in retinal photoreceptor cells. Neuroscience 11:857–866

    Google Scholar 

  • Kreutzberg GW, Barron KD, Schubert P (1978) Cytochemical localization of 5′-nucleotidase in glial plasma membranes. Brain Res 158:247–257

    Google Scholar 

  • Kreutzberg GW, Heymann D, Reddington M (1986a) 5′-nucleotidase in the nervous system. In: Kreutzberg GW, Reddington M, Zimmermann H (eds) Cellular biology of ectoenzymes. Springer, Berlin Heidelberg New York, pp 147–164

    Google Scholar 

  • Kreutzberg GW, Reddington M, Zimmermann H (eds) (1986b) Cellular biology of ectoenzymes. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Leranth C, Feher E (1983) Synaptology and sources of vasoactive intestinal polypeptide and substance P containing axons of the cat celiac ganglion. An experimental electron microscopic immunohistochemical study. Neuroscience 10:947–958

    Google Scholar 

  • Lee KS, Schubert P, Emmert H, Kreutzberg GW (1981) The effects of adenosine versus adenine nucleotides on evoked potentials in a rat hippocampal slice preparation. Neurosci Lett 23:309–314

    Google Scholar 

  • Lee KS, Schubert P, Reddington M, Kreutzberg GW (1986) The distribution of adenosine Al receptors and 5′-nucleotidase in the hippocampal formation of several mammalian species. J Comp Neurol 246:427–434

    Google Scholar 

  • Luzio JP, Bailyes EM, Baron M, Siddle K, Mullock BM, Geuze HJ, Stanley KK (1986) The properties, structure, function, intracellular localisation and movement of hepatic 5′-nucleotidase. In: Kreutzberg GW, Reddington M, Zimmerman H (eds) Cellular biology of ectoenzymes. Springer, Berlin Heidelberg New York, pp 89–116

    Google Scholar 

  • Marani E (1977) The subcellular distribution of 5′-nucleotidase activity in mouse cerebellum. Exp Neurol 57:1042–1048

    Google Scholar 

  • Mescher MF, Jose MJL, Balk SP (1981) Actin-containing matrix associated with the plasma membrane of murine tumour and lymphoid cells. Nature 289:139–144

    Google Scholar 

  • Montero VM, Singer W (1984) Ultrastructure and synaptic relations of neural elements containing glutamic acid decarboxylase (GAD) in the perigeniculate nucleus of the cat. Exp Brain Res 56:115–125

    Google Scholar 

  • Naidoo D (1962) The activity of 5′-nucleotidase determined histochemically in the developing rat brain. J Histochem Cytochem 10:421–434

    Google Scholar 

  • Oertel WH, Schmechel DE, Mugnaini E, Tappaz ML, Kopin IJ (1981) Immunocytochemical localization of glutamate decarboxylase in rat cerebellum with a new antiserum. Neuroscience 6:2715–2735

    Google Scholar 

  • Pasik P, Pasik T, Saavedra JP (1982) Immunocytochemical localization of serotonin at the ultrastructural level. J Histochem Cytochem 30:760–764

    Google Scholar 

  • Peters A, Palay SL, Webster H deF (1976) The fine structure of the nervous system: The neurons and supporting cells. WB Saunders, Philadelphia London Toronto

    Google Scholar 

  • Schachner M, Hedley-White ET, Hsu DW, Schoonmaker G, Bignami A (1977) Ultrastructural localization of glial fibrillary acidic protein in mouse cerebellum by immunoperoxidase labeling. J Cell Biol 75:67–73

    Google Scholar 

  • Scott TG (1965) The specificity of 5′-nucleotidase in the brain of the mouse. J Histochem Cytochem 13:657–667

    Google Scholar 

  • Scott TG (1967) The distribution of 5′-nucleotidase in the brain of the mouse. J Comp Neurol 129:97–114

    Google Scholar 

  • Siddle K, Bailyes EM, Luzio JP (1981) A monoclonal antibody inhibiting rat liver 5′-nucleotidase. FEBS Lett 128:103–107

    Google Scholar 

  • Somogyi P, Hodgson AJ, Chubb IW, Penke B, Erdei A (1985) Antiserum to gamma-aminobutyric acid: II. Immunocytochemical application to the central nervous system. J Histochem Cytochem 33:240–248

    Google Scholar 

  • Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333

    Google Scholar 

  • Stefanovic V, Mandel P, Rosenberg A (1976) Ecto-5′-nucleotidase of intact cultured C6 rat glioma cells. J Biol Chem 251:3900–3905

    Google Scholar 

  • Uusitalo RJ, Karnovsky MJ (1977) Surface localization of 5′-nucleotidase on the mouse lymphocyte. J Histochem Cytochem 25:87–96

    Google Scholar 

  • Wachstein M, Meisel E (1957) Histochemistry of hepatic phosphatases at a physiologic pH with special reference to the demonstration of bile canaliculi. Am J Clin Pathol 27:13–23

    Google Scholar 

  • Widnell CC (1972) Cytochemical localization of 5′-nucleotidase in subcellular fractions isolated from rat liver I. The origin of 5′-nucleotidase activity in microsomes. J Cell Biol 52:542–558

    Google Scholar 

  • Widnell CC, Schneider YJ, Pierre B, Baudhuin P, Trouet A (1982) Evidence for a continual exchange of 5′-nucleotidase between the cell surface and cytoplasmic membranes in cultured rat fibroblasts. Cell 28:61–70

    Google Scholar 

  • Yamashina S, Kawai K (1979) Cytochemical studies on the localization of 5′-nucleotidase in the acinar cells of the rat salivary glands. Histochemistry 60:255–236

    Google Scholar 

  • Zachowski A, Evans WH, Paraf A (1981) Immunological evidence that plasma membrane 5′-nucleotidase is a transmembrane protein. Biochim Biophys Acta 644:121–126

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neuromorphology, Max-Planck-Institute for Psychiatry, Am Klopferspitz 18a, D-8033, Martinsried, Germany

    S. W. Schoen, M. B. Graeber, M. Reddington & G. W. Kreutzberg

Authors
  1. S. W. Schoen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. B. Graeber
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Reddington
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. W. Kreutzberg
    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

Schoen, S.W., Graeber, M.B., Reddington, M. et al. Light and electron microscopical immunocytochemistry of 5′-nucleotidase in rat cerebellum. Histochemistry 87, 107–113 (1987). https://doi.org/10.1007/BF00533394

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00533394

Keywords

Search

Navigation