Summary
Small amounts of labeled gamma-aminobutyric acid (3H-GABA), a putative neurotransmitter, were injected stereotaxically into the cerebral and cerebellar cortices of rats pretreated with aminooxyacetic acid (AOAA), a drug which prevents GABA breakdown. After fixation by aldehyde perfusion the tissue was processed for light and electron microscopic autoradiography.
Specific autoradiographic uptake patterns were observed in both cortices. A high activity was found mainly over stellate and basket cells in the cerebellar cortex and over stellate cells in the cerebral cortex. Also probable Golgi cells seem to accumulate 3H-GABA. Pyramidal cells and Purkinje cells, on the other hand, showed a low activity. Thus, uptake and retention of 3H-GABA in the brain areas studied seem to be related to interneurons supposed to have an inhibitory function. This is consistent with the view that GABA serves as the major inhibitory transmitter substance in mammalian cortical areas.
Similar content being viewed by others
References
Aghajanian, G.K., Bloom, F.E.: Electron microscopic autoradiography of rat hypothalamus after intraventricular H3-norepinephrine. Science 153, 308–310 (1966).
Bell, C.C., Dow, C.S.: Cerebellar circuitry. Neurosci. Res. Prog. Bull. 5, 121–222 (1967).
Bloom, F.E., Iversen, L.L.: Localizing 3H-GABA in nerve terminals of rat cerebral cortex by electron microscopic autoradiography. Nature (Lond.) 229, 628–630 (1970).
Caro, L., Van Tubergen, R.P.: High resolution autoradiography. I. Methods. J. Cell Biol. 15, 173–188 (1962).
Colonnier, M.L.: The structural design of the neocortex. In: Brain and conscious experience. Ed. by J.C. Eccles. New Jersey: Springer 1966.
—: Synaptic patterns on different cell types in the different laminae of the cat visual cortex. An electron microscope study. Brain Res. 9, 268–287 (1968).
Curtis, D.R., Johnston, G.A.R.: Amino acid transmitters. In: Handbook of Neurochemistry. Ed. by A. Lajtha. New York-London: Plenum Press 1970.
Descarrier, L., Droz, B.: Intraneural distribution of exogenous norepinephrine in the central nervous system of the rat. J. Cell Biol. 44, 385–399 (1970).
Eccles, J.C., Ito, M., Szentágothai, J.: The cerebellum as a neuronal machine. Berlin-Heidelberg-New York: Springer 1967.
Ehinger, B.: Autoradiographic identification of rabbit retintal neurons that take up GABA. Experientia (Basel) 26, 1063 (1970).
— Falck, B.: Autoradiography of GABA, glycine, and some other suspected neurotransmitter substances. Brain Res. 33, 157–172 (1971).
Faeder, I.R., Salpeter, M.M.: Glutamate uptake by a stimulated insect nerve muscle preparation. J. Cell Biol. 46, 303–307 (1970).
Fonnum, F., Storm-Mathisen, J., Walberg, F.: Glutamate decarboxylase in inhibitory neurons. A study of the enzyme in Purkinje cell axons and boutons in the cat. Brain Res. 20, 259–276 (1970).
Fox, C.A., Hillman, D.E., Siegesmund, K.A., Dutta, C.R.: The primate cerebellar cortex: A Golgi and electron microscopic study. In: The Cerebellum. Ed. by C.A. Fox and R.S. Snider. Progress in Brain Research, Vol. 25, 174–225, Elsevier 1967.
Gerschenfeld, H.M., Wald, F., Zadunaisky, J.A., De Robertis, E.: Function of astroglia in the water ion metabolism of the central nervous system. Neurology 9, 421–425 (1959).
Henn, F.A., Hamberger, A.: Glial cell function. Uptake of transmitter substances. Proc. nat. Acad. Sci. (Wash.) 68, 2686–2690 (1971).
Hebb, C.: CNS at the cellular level: Identity of transmitter agents. Ann. Rev. Physiol. 32, 165–192 (1970).
Hökfelt, T.: In vitro studies on central and peripheral monoamine neurons at the ultrastructural level. Z. Zellforsch. 91, 1–74 (1968).
— Ljungdahl, Å.: Cellular localization of labeled gamma-aminobutyric acid (3H-GABA) in rat cerebellar cortex: an autoradiographic study. Brain Res. 22, 391–396 (1970).
—: Light and electron microscopic autoradiography on spinal cord slices after incubation with labeled glycine. Brain Res. 32, 189–194 (1971a).
Hökfelt, T. Ljungdahl, Å.: Uptake of 3H-noradrenaline and 3H-gamma-aminobutyric acid in isolated tissues of rat: An autoradiographic and fluorescence microscopic study. In: Progress in Brain Research, Histochemistry of Nervous Transmission. Ed. by O. Eränkö. Progress in Brain Research, Vol. 34, 87–102, Elsevier 1971b.
—: Histochemical determination of neurotransmitter distribution. Proceedings of the Association for Research in Nervous and Mental Diseases, Williams and Wilkins, Publishers, Conference on Neurotransmitters, December 4 and 5, 1970, New York (in press) (1971c).
Hökfelt, T. Ljungdahl, Å.: Application of cytochemical technique to the study of suspect transmitter substances in the nervous system. In: Advances in Biochemical Psychopharmacology. Ed. by E. Costa and L.L. Iversen. (in press) (1971d).
Iversen, L.L., Neal, M.J.: The uptake of (3H) GABA by slices of rat cerebral cortex. J. Neurochem. 15, 1141–1149 (1968).
— Snyder, S.H.: Synaptosomes: Different populations storing catecholamines and gamma-aminobutyric acid in homogenates of rat brain. Nature (Lond.) 220, 796–798 (1968).
Krnjević, K., Schwartz, S.: Some properties of unresponsive cells in the cerebral cortex. Exp. Brain Res. 3, 306–319 (1967).
—: The action of γ-aminobutyric acid on cortical neurons. Exp. Brain Res. 3, 320–336 (1967).
Luft, J.H.: Improvements in epoxy resin embedding methods. J. biophys. biocliem. Cytol. 9, 409–414 (1961).
Miller, O.L., jr., Stone, G.E., Prescott, D.M.: Autoradiography of soluble materials. J. Cell Biol. 23, 654–658 (1964).
Nagata, T., Nawa, T.: A modification of dry-mounting technique for radioautography of water-soluble compounds. Histoehemie 7, 370–371 (1966).
Neal, M.J., Iversen, L.L.: Subcellular distribution of endogenous and [3H]γ-aminobutyric acid in rat cerebral cortex. J. Neurochem. 16, 1245–1252 (1969).
Obata, K., Ito, M., Ochi, R., Sato, N.: Pharmacological properties of the postsynaptic inhibition by Purkinje cell axons and the action of γ-aminobutyric acid on Deiters neurones. Exp. Brain Res. 4, 43–57 (1967).
— Otsuka, M., Tanaka, Y.: Determination of gamma-aminobutyric acid in single nerve cells of cat central nervous system. J. Neurochem. 17, 697–698 (1970).
Orkand, P.M., Kravitz, E.A.: Localization of the sites of γ-aminobutyric acid (GABA) uptake in lobster nerve-muscle preparations. J. Cell Biol. 49, 75–89 (1971).
Peters, A.: Stellate cells of the rat parietal cortex. J. comp. Neurol. 141, 345–374 (1971).
Reynolds, E.S.: The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol. 17, 208–212 (1963).
Scholl, D.A.: The organization of the cerebral cortex. London: Methuen 1956.
Schultz, R.L., Case, N.M.: A modified aldehyde perfusion technique for preventing certain artifacts in electron microscopy of the central nervous system. J. Microscopy 92, 69–84 (1970).
Thorack, R.M., Dufty, M.L., Haynes, J.M.: The effect of anisotonic media upon cellular ultrastructure in fresh and fixed rat brain. Z. Zellforsch. 66, 690–700 (1965).
Wallach, D.P.: Studies on the GABA pathway. The inhibition of γ-aminobutyric acid-α-ketoglutaric acid transaminase in vitro and in vivo by U7524 (amino-oxyacetic acid). Biochem. Pharmacol. 5, 323–331 (1961).
Wolfe, D.E., Potter, L.T., Richardson, K.C., Axelrod, J.: Localizing tritiated norepinephrine in sympathetic axons by electron microscope autoradiography. Science 138, 440–442 (1962).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hökfelt, T., Ljungdahl, Å. Autoradiographic identification of cerebral and cerebellar cortical neurons accumulating labeled gamma-aminobutyric acid (3H-GABA). Exp Brain Res 14, 354–362 (1972). https://doi.org/10.1007/BF00235032
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00235032