Summary
Sections of fixed, paraffin-embedded brain from mice and rats were stained with agglutinin fromVicia villosa conjugated to horseradish peroxidase (VVA-HRP) to localize glycoconjugate containing terminalN-acetylgalactosamine (GalNAc). VVA-HRP binding sites were localized in periodic foci at the surface of a selective population of non-pyramidal interneurons in layers II through VI of the rodent cerebral cortex. These multipolar interneurons were shown to utilize γ-aminobutyric acid (GABA) as a transmitter and thus to be GABAergic by their immunostaining for glutamic acid decarboxylase (GAD) throughout the cytoplasm in serial sections. Pyramidal and other non-pyramidal cortical neurons received GABAergic input as evidenced by punctate immunostaining for GAD on their soma and proximal dendritic aborizations, but these cells failed to show VVA affinity or cytosolic GAD reactivity. Most neurons in the thalamic reticular nucleus stained for the presence of glycoconjugate with terminal GalNAc on their surface and for GAD in the cytosol. In contrast, cerebellar Purkinje cells showed strong cytosolic reactivity with anti-GAD but lacked surface staining with VVA-HRP. These observations show that some but not other populations of GABAergic neurons possess binding capacity for VVA on their surface. The surface of neurons in the deep cerebellar nucleus stained heavily with VVA but failed to show clear cytosolic reactivity for GAD. Some neurons with surface glycoconjugate containing terminal GalNAc are therefore not GABAergic.
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References
Chan-Palay, V. (1978) Autoradiographic localization of (gamma-aminobutyric acid) receptors in the rat central nervous system by using [3H] muscimol.Proceedings of the National Academy of Sciences USA 75, 1024–8.
Chan-Palay, V., Palay, S. L. &Wu, J.-Y. (1979) Gamma-aminobutyric acid pathways in the cerebellum studied by retrograde and anterograde transport of glutamic acid decarboxylase antibody afterin vivo injections.Anatomy and Embryology 157, 1–14.
Feldman, M. L. &Peters, A. (1978) The forms of non-pyramidal neurons in the visual cortex of the rat.Journal of Comparative Neurology 179, 761–94.
Hendrickson, A. E., Hunt, S. &Wu, J.-Y. (1981) Immunocytochemical localization of glutamic acid decarboxylase in monkey striate cortex.Nature 292, 605–7.
Hendry, S. H. C., Houser, C. R., Jones, E. G. &Vaughn, J. E. (1983) Synaptic organization of immunocytochemically identified GABA neurons in the monkey sensory-motor cortex.Journal of Neurocytology 12, 639–60.
Houser, C. R., Hendry, S. H. C., Jones, E. G. &Vaughn, J. E. (1983) Morphological diversity of immunocytochemically defined GABA neurons in the monkey sensory-motor cortex.Journal of Neurocytology 12, 617–38.
Houser, C. R., Vaughn, J. E., Barber, R. P. &Roberts, E. (1980) GABA neurons are the major cell type of the nucleus reticularis thalami.Brain Research 200, 341–54.
Ledeen, R. W. (1978) Ganglioside structures and distribution: are they localized at the nerve ending?Journal of Supramolecular Structure 8, 1–17.
Margolis, R. U. &Margolis, R. K. (1977) Metabolism and function of glycoproteins and glycosaminoglycans in nervous tissue.International Journal of Biochemistry 8, 85–91.
McLaughlin, B. J., Wood, J. G., Saito, K., Barger, R., Vaughn, J. E., Roberts, E. &Wu, J.-Y. (1974) The fine structural localization of glutamate decarboxylase in synaptic terminals of rodent cerebellum.Brain Research 76, 377–91.
Nakagawa, F., Schulte, B. A. &Spicer, S. S. (1986) Selective cytochemical demonstration of glycoconjugate containing terminal N-acetylgalactosamine in some brain neurons.Journal of Comparative Neurology 243, 280–90.
Oertel, W. H., Mugnaini, E., Schmechel, D. E., Tappaz, M. L. &Kopin, I. J. (1982) The immunocytochemical demonstration of gamma-aminobutyric acidergic neurons — methods and application. InCytochemical Methods in Neuroanatomy (edited byChan-Palay, V. &Palay, S. L.), pp. 297–329. New York: Alan R. Liss.
Ottersen, O. P. &Storm-Mathisen, J. (1984) Glutamate- and GABA-containing neurons in the mouse and rat brain, as demonstrated with a new immunocytochemical technique.Journal of Comparative Neurology 229, 374–92.
Palay, S. L. &Chan-Palay, V. (1975) Synaptic analysis of the neuropil.Cold Spring Harbor Symposium of Quantitative Biology 40, 1–16.
Ribak, C. E. (1978) Aspinous and sparsely-spinous stellate neurons in the visual cortex of rats contain glutamic acid decarboxylase.Journal of Neurocytology 7, 461–78.
Ribak, C. E., Vaughn, J. E. &Saito, K. (1978) Immunocytochemical localization of glutamic acid decarboxylase in neuronal somata following colchicine inhibition of axonal transport.Brain Research 140, 315–32.
Saito, K., Barber, R., Wu, J.-Y., Matsuda, T., Roberts, E. &Vaughn, J. E. (1974) Immunohistochemical localization of glutamic acid decarboxylase in rat cerebellum.Proceedings of the National Academy of Sciences USA 71, 269–73.
Schulte, B. A. &Spicer, S. S. (1983) Light microscopic detection of sugar residues in glycoconjugates of salivary glands and the pancreas with lectin-horseradish peroxidase conjugates. I. Mouse.Histochemical Journal 15, 1217–38.
Somogyi, P., Freund, T., Wu, J.-Y. &Smith, A. D. (1983) The section Golgi impregnation procedure. II. Immunocytochemical demonstration of glutamate decarboxylase in Golgi-impregnated neurons and in their afferent and efferent synaptic boutons in the visual cortex of the cat.Neuroscience 9, 475–90.
Somogyi, P., Hodgson, A. J., Chubb, I. W., Penke, B., &Erdei, A. (1985) Antisera to γ-aminobutyric acid. II. Immunocytochemical application to the central nervous system.Journal of Histochemistry and Cytochemistry 33, 240–8.
Tollefsen, S. E. &Kornfeld, R. (1983a) Isolation and characterization of lectins fromVicia villosa.Journal of Biological Chemistry 258, 5165–71.
Tollefsen, S. E. &Kornfeld, R. (1983b) The B4 lectin fromVicia villosa seeds interacts with N-acetylgalactosamine residues α-linked to serine or threonine residues in cell surface glycoproteins.Journal of Biological Chemistry 258, 5172–6.
Wu, J.-Y. (1983) Preparation of glutamic acid decarboxylase as immunogen for immunocytochemistry. InNeuroimmunocytochemistry (IBRO Handbook Series: Methods in the Neurosciences) (edited byCuello, A. C.), pp. 159–91. Sussex: John Wiley & Sons.
Wu, J.-Y., Lin, C.-T., Brandon, C., Chan, D.-S., Möhler, H. &Richards, J. G., (1982) Regulation and immunocytochemical characterization of GAD. InCytochemical Methods in Neuroanatomy (edited byPalay, S. L. &Chan-Palay, V.), pp. 279–96. New York: Alan R. Liss.
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Nakagawa, F., Schulte, B.A., Wu, J.Y. et al. GABAergic neurons of rodent brain correspond partially with those staining for glycoconjugate with terminalN-acetylgalactosamine. J Neurocytol 15, 389–396 (1986). https://doi.org/10.1007/BF01611440
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DOI: https://doi.org/10.1007/BF01611440