The Histochemical Journal

, Volume 33, Issue 9–10, pp 537–543

GABA Immunoreactivity in the Human Cerebellar Cortex: A Light and Electron Microscopical Study

  • Vincenzo Benagiano
  • Luisa Roncali
  • Daniela Virgintino
  • Paolo Flace
  • Mariella Errede
  • Anna Rizzi
  • Francesco Girolamo
  • David Robertson
  • Joachim Bormann
  • Glauco Ambrosi
Article

Abstract

The distribution of γ-aminobutyric acid (GABA) in surgical samples of human cerebellar cortex was studied by light and electron microscope immunocytochemistry using a polyclonal antibody generated in rabbit against GABA coupled to bovine serum albumin with glutaraldehyde. Observations by light microscopy revealed immunostained neuronal bodies and processes as well as axon terminals in all layers of the cerebellar cortex. Perikarya of stellate, basket and Golgi neurons showed evident GABA immunoreactivity. In contrast, perikarya of Purkinje neurons appeared to be negative or weakly positive. Immunoreactive tracts of longitudinally- or obliquely-sectioned neuronal processes and punctate elements, corresponding to axon terminals or cross-sectioned neuronal processes, showed a layer-specific pattern of distribution and were seen on the surface of neuronal bodies, in the neuropil and at microvessel walls. Electron microscope observations mainly focussed on the analysis of GABA-labelled axon terminals and of their relationships with neurons and microvessels. GABA-labelled terminals contained gold particles associated with pleomorphic vesicles and mitochondria and established symmetric synapses with neuronal bodies and dendrites in all cortex layers. GABA-labelled terminals associated with capillaries were seen to contact the perivascular glial processes, basal lamina and endothelial cells and to establish synapses with subendothelial unlabelled axons.

To our Master, Professor Rodolfo Amprino, with our great admiration, gratefulness and affection, on the occasion of his ninetieth birthday.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Benagiano V, Virgintino D, Rizzi A, Flace P, Troccoli V, J, Monaghan P, Robertson D, Roncali L, Ambrosi G (2000a) Glutamic acid decarboxylase-positive neuronal cell bodies and terminals in the human cerebellar cortex. Histochem J 32: 557–564.Google Scholar
  2. Benagiano V, Flace P, Virgintino D, Rizzi A, Roncali L, Ambrosi G (2000b) Immunolocalization of glutamic acid decarboxylase in postmortem human cerebellar cortex.Alight microscopy study. Histochem Cell Biol 114: 191–195.Google Scholar
  3. Bishop GA, Chen YF, Burry RW, King JS (1993) An analysis of GABAergic afferents to basket cell bodies in the cat's cerebellum. Brain Res 623: 293–298.Google Scholar
  4. Carlemalm E, Garavito RM, Villiger W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc 126: 123–143.Google Scholar
  5. Erdo SL, Joo F, Wolff JR (1989) Immunohistochemical localization of glutamate decarboxylase in the rat oviduct and ovary: Further evidence for non-neural GABA systems. Cell Tissue Res 255: 431–434.Google Scholar
  6. Gabbott PL, Somogyi J, Stewart MG, Hamori J (1986) GABAimmunoreactive neurons in the rat cerebellum: A light and electron microscope study. J Comp Neurol 251: 474–490.Google Scholar
  7. Gilon P, Campistron G, Geffard M, Remacle C (1988) Immunocytochemical localisation of GABA in endocrine cells of the rat enteropancreatic system. Biol Cell 62: 265–273.Google Scholar
  8. Gragera RR, Muniz E, Martinez-Rodriguez R (1993) Electron microscopic immunolocalization ofGABAand glutamic acid decarboxylase in cerebellar capillaries and their microenvironment. Cell Mol Biol 39: 809–817.Google Scholar
  9. Hamori J, Takacs J, Petrusz P (1990) Immunogold electron microscopic demonstration of glutamate and GABA in normal and deafferented cerebellar cortex: Correlation between transmitter content and synaptic vesicle size. J Histochem Cytochem 38: 1767–1777.Google Scholar
  10. Hodgson A, Penke B, Erdei A, Chubb IW, Somogyi P (1985) Antiserum to γ-aminobutyric acid: I. Preparation and characterization using a new model system. J Histochem Cytochem 33: 229–239.Google Scholar
  11. Imai H, Okuno T, Wu JY, Lee TJ (1991) GABAergic innervation in cerebral blood vessels: An immunohistochemical demonstration of L-glutamic acid decarboxylase and GABA transaminase. J Cereb Blood Flow Metab 11: 129–134.Google Scholar
  12. Martin DL, Tobin AJ (2000) Mechanisms controlling GABA synthesis and degradation in the brain. In: Martin DL, Olsen RW, eds. GABA in the Nervous System: The View at Fifty Years. Philadelphia: Lippincott Williams & Wilkins, pp. 25–41.Google Scholar
  13. Martinez-Rodriguez R, Tonda A, Gragera RR, Paz-Doel R, Garcia-Cordovilla R, Fernandez-Fernandez E, Fernandez AM, Gonzalez-Romero F, Lopez Bravo A(1993) Synaptic and non-synaptic immunolocalization of GABA and glutamate acid decarboxylase (GAD) in cerebellar cortex of rat. Cell Mol Biol 39: 115–123.Google Scholar
  14. McLaughlin BJ, Wood JG, Saito K, Barber R, Vaughn JE, Roberts E, Wu J-Y (1974) The fine structural localization of glutamate decarboxylase in synaptic terminals of rodent cerebellum. Brain Res 76: 377–391.Google Scholar
  15. Miranda FJ, Torregrosa G, Salom JB, Campos V, Alabadi JA, Alborch E (1989) Inhibitory effect ofGABAon cerebrovascular sympathetic neurotransmission. Brain Res 492: 45–52.Google Scholar
  16. Monaghan P, Robertson D, Beesley JE (1993) Immunolabelling techniques for electron microscopy. In Beesley JE, ed. Immunocytochemistry: A Practical Approach, Oxford: University Press, pp. 43–76.Google Scholar
  17. Mugnaini E (2000) GABAergic inhibition in the cerebellar system. In: Martin DL, Olsen RW, eds. GABA in the Nervous System: The View at Fifty Years. Philadelphia: Lippincott Williams & Wilkins, pp. 383–407.Google Scholar
  18. Mugnaini E, Oertel WH (1985) An atlas of the distribution ofGABAergic neurons and terminals in the rat CNS as revealed by GAD immunohistochemistry. In: Björklund A, Hökfelt T, eds. Handbook of Chemical Neuroanatomy, Vol. 4. GABA and Neuropeptides in the CNS-Part I. British-Vancouver: Elsevier, pp. 436–608.Google Scholar
  19. Oertel WH, Schmechel DE, Mugnaini E, Tappaz ML, Kopin IJ (1981) Immunocytochemical localisation of glutamate decarboxylase in rat cerebellum with a new antiserum. Neuroscience 6: 2715–2735.Google Scholar
  20. Ottersen OP, Madsen S, Storm-Mathisen J, Somogyi P, Scopsi L, Larsson LI (1988) Immunocytochemical evidence suggests that taurine is colocalized with GABA in the Purkinje cell terminals, but that the stellate cell terminals predominantly contain GABA: A light and electron microscopic study of the rat cerebellum. Exp Brain Res 72: 407–416.Google Scholar
  21. Peters A, Palay SL, Webster HD(1991) Synapses. In: Peters A, Palay SL, Webster HD, eds. The Fine Structure of the Nervous System.NewYork, Oxford: University Press, pp. 138–211.Google Scholar
  22. Ribak CE, Vaughn JE, Saito K (1978) Immunocytochemical localization of glutamic acid decarboxylase in neuronal somata following colchicine inhibition of axonal transport. Brain Res 140: 315–332.Google Scholar
  23. Robertson D, Monaghan P, Clarke C, Atherton A (1992) An appraisal of low-temperature embedding by progressive lowering of temperature into Lowicryl HM20 for immunocytochemical studies. J Microsc 168: 85–100.Google Scholar
  24. Saito K, Barber R, Wu J-Y, Matsuda T, Roberts E, Vaughn JE (1974) Immunohistochemical localization of glutamate decarboxylase in rat cerebellum. Proc Natl Acad Sci USA 71: 269–273.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Vincenzo Benagiano
    • 1
    • 2
  • Luisa Roncali
    • 2
  • Daniela Virgintino
    • 2
  • Paolo Flace
    • 2
  • Mariella Errede
    • 2
  • Anna Rizzi
    • 2
  • Francesco Girolamo
    • 2
  • David Robertson
    • 3
  • Joachim Bormann
    • 4
  • Glauco Ambrosi
    • 5
  1. 1.Facoltà di Medicina e ChirurgiaUniversità di FoggiaItaly
  2. 2.Dipartimento di Anatomia Umana e IstologiaUniversità di BariItaly
  3. 3.Haddow LaboratoriesInstitute of Cancer ResearchSuttonUK
  4. 4.Lehrstul für ZellphysiologieRuhr-UniversitätBochumGermany
  5. 5.Dipartimento di Anatomia Umana e IstologiaUniversità di BariItaly

Personalised recommendations