Summary
The coexistence of immunoreactivities for choline acetyltransferase (ChAT) and glutamic acid decarboxylase (GAD) and/or gamma-aminobutyric acid (GABA) was revealed in some brain regions of the rat, using the peroxidase-antiperoxidase method. Consecutive 40 μm thick vibratome sections were incubated in different antisera and those cells which were bisected by the plane of sectioning so as to be included at the paired surfaces of two adjacent sections were identified. The coexistence of the immunoreactivities for ChAT and GAD or GABA in the same cell could thus be determined by observing the immunoreactivity of the two halves of the cell incubated in two different antisera. In the retina, cerebral cortex, basal forebrain and spinal cord, colocalization of ChAT-like and GAD-like or GABA-like immunoreactivities was observed in some cell types, whereas no such colocalization was observed in cells in the striatum or brainstem. In the retina, the majority of ChAT-like immunoreactive (ChAT-LI) amacrine cells contained GABA-like or GAD-like immunoreactivity. About half of the ChAT-LI neurons in the cerebral cortex showed GABA-like immunoreactivity. In the basal forebrain only a small proportion of ChAT-LI neurons (0.6%) contained GAD-like immunoreactivity. In the spinal cord, about one-third of ChAT-LI central canal cluster cells and about half of ChAT-LI dorsal horn cells showed GAD-like and/or GABA-like immunoreactivities. These observations indicate the possible coexistence of two classical transmitters, GABA and acetylcholine, in various brain regions and spinal cord of the rat.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ariel M, Daw NW (1982) Pharmacological analysis of directionally sensitive rabbit retinal ganglion cells. J Physiol 324: 161–185
Armstrong DM, Saper CB, Levey AI, Wainer BH, Terry RD (1983) Distribution of cholinergic neurons in rat brain: demonstrated by the immunocytochemical localization of choline acetyltransferase. J Comp Neurol 216: 53–68
Barber RP, Phelps PE, Houser CR, Crawford GD, Salvaterra PM, Vaughn JE (1984) The morphology and distribution of neurons containing choline acetyltransferase in the adult rat spinal cord: an immunocytochemical study. J Comp Neurol 229: 329–346
Barber RP, Vaughn JE, Roberts E (1982) The cytoarchitecture of GABAergic neurons in rat spinal cord. Brain Res 238: 305–328
Belin MF, Nanopoulos D, Didier M, Aguera M, Steinbusch H, Verhofstad A, Maitre M, Pujol JF (1983) Immunohistochemical evidence for the presence of γ-aminobutyric acid and serotonin in one nerve cell. A study on the raphe nuclei of the rat using antibodies to glutamate decarboxylase and serotonin. Brain Res 275: 329–339
Black IB, Adler JE, Dreyfus CF, Jonakait GM, Katz DM, La-Gamma EF, Markey KM (1984) Neurotransmitter plasticity at the molecular level. Science 225: 1226–1270
Brandon C (1985) Retinal GABA neurons: localization in vertebrate species using an antiserum to rabbit brain glutamate decarboxylase. Brain Res 344: 286–295
Brashear HR, Zaborszky L, Heimer L (1986) Distribution of GABAergic and cholinergic neurons in the rat diagonal band. Neuroscience 17: 439–451
Chan-Palay V, Engel AG, Palay SL, Wu J-Y (1982a) Synthesizing enzymes for four neuroactive substances in motor neurons and neuromuscular junctions: light and electron microscopic immunocytochemistry. Proc Natl Acad Sci USA 79: 6717–6721
Chan-Palay V, Engel AG, Wu J-Y, Palay SL (1982b) Coexistence in human and primate neuromuscular junctions of enzymes synthesizing acetylcholine, catecholamine, taurine, and γ-aminobutyric acid. Proc Natl Acad Sci USA 79: 7027–7030
Eckenstein F, Baughman RW (1984) Two types of cholinergic innervation in cortex, one co-localized with vasoactive intestinal polypeptide. Nature 309: 153–155
Eckenstein F, Thoenen H (1983) Cholinergic neurons in the rat cerebral cortex demonstrated by immunohistochemical localization of choline acetyltransferase. Neurosci Lett 36: 211–215
Everitt BJ, Hökfelt T, Wu J-Y, Goldstein M (1984) Coexistence of tyrosine hydroxylase-like and gamma-aminobutyric acid-like immunoreactivities in neurons of the arcuate nucleus. Neuroendocrinology 39: 189–191
Harandi M, Aguera M, Gamrani H, Didier M, Maitre M, Calas A, Belin MF (1987) γ-Aminobutyric acid and 5-hydroxytryptamine interrelationship in the rat nucleus raphe dorsalis: combination of radioautographic and immunocytochemical techniques at light and electron microscopy levels. Neuroscience 21: 237–251
Henderson Z (1987) A small proportion of cholinergic neurones in the nucleus basalis magnocellularis of ferret appear to stain positively for tyrosine hydroxylase. Brain Res 412: 363–369
Houser CR, Crawford GD, Barber RP, Salvaterra PM, Vaughn JE (1983) Organization and morphological characteristics of cholinergic neurons: an immunocytochemical study with a monoclonal antibody to choline acetyltransferase. Brain Res 266: 97–119
Houser CR, Crawford GD, Salvaterra PM, Vaughn JE (1985) Immunocytochemical localization of choline acetyltransferase in rat cerebral cortex: a study of cholinergic neurons and synapses. J Comp Neurol 234: 17–34
Ichikawa T, Hirata Y (1986) Organization of choline acetyltransferase-containing structures in the forebrain of the rat. J Neurosci 6:281–292
Johnson CD, Epstein ML (1986) Monoclonal antibodies and polyvalent antiserum to chicken choline acetyltransferase. J Neurochem 46: 968–976
Kimura K, McGeer PL, Peng J-H (1984) Choline acetyltransferase-containing neurons in the rat brain. In: Björklund A, Hökfelt T, Kuhar MJ (eds) Classical transmitters and transmitter receptors in the CNS, part II. Handbook of chemical neuroanatomy, Vol 3. Elsevier/North-Holland, Amsterdam, pp 51–67
Kosaka T, Hataguchi Y, Kama K, Nagatsu I, Wu J-Y (1985a) Coexistence of immunoreactivities for glutamate decarboxylase and tyrosine hydroxylase in some neurons in the periglomerular region of the rat main olfactory bulb: possible coexistence of gamma-aminobutyric acid (GABA) and dopamine. Brain Res 343: 166–171
Kosaka T, Kosaka K, Tateishi K, Hamaoka Y, Yanaihara N, Wu J-Y, Hama K (1985b) GABAergic neurons containing CCK-8-like and/or VIP-like immunoreactivities in the rat hippocampus and dentate gyrus. J Comp Neurol 239: 420–430
Kosaka T, Kosaka K, Hataguchi Y, Nagatsu I, Wu J-Y, Ottersen OP, Storm-Mathisen J, Hama K (1987a) Catecholaminergic neurons containing GABA-like and/or glutamic acid decarboxylase-like immunoreactivities in various brain regions of the rat. Exp Brain Res 66: 191–210
Kosaka K, Hama K, Nagatsu I, Wu J-Y, Ottersen OP, Storm-Mathisen J, Kosaka T (1987b) Postnatal development of neurons containing both Catecholaminergic and GABAergic traits in the rat main olfactory bulb. Brain Res 403: 355–360
Levey AI, Wainer BH, Rye DB, Mufson EJ, Mesulam M-M (1984) Choline acetyltransferase-immunoreactive neurons intrinsic to rodent cortex and distinction from acetylcholineserase-positive neurons. Neuroscience 13: 341–353
Masland RH, Tauchi M (1986) The cholinergic amacrine cell. Trends Neurosci 9: 218–223
Massey SC, Neal MJ (1979) The light evoked release of acetylcholine from rabbit retina in vivo and its inhibition by γ-aminobutyric acid. J Neurochem 32: 1327–1329
Mesulam M-M, Mufson EJ, Wainer BH, Levey AI (1983) Central cholinergic pathways in the rat: an overview based on an alternative nomenclature (Ch1-Ch6). Neuroscience 10: 1185–1201
Miceli MO, Post CA, van der Kooy D (1987) Catecholamine and serotonin colocalization in projection neurons of the area postrema. Brain Res 412: 381–385
Millhorn DE, Hökfelt T, Seroogy K, Oertel W, Verhofstad AAJ, Wu J-Y (1987) Immunohistochemical evidence for colocalization of γ-aminobutyric acid and serotonin in neurons of the ventral medulla oblongata projecting to the spinal cord. Brain Res 410: 179–185
Mosinger J, Yazulla S (1987) Double-label analysis of GAD-and GABA-like immunoreactivity in the rabbit retina. Vision Res 27: 23–30
Mugnaini E, Oertel WH (1985) An atlas of the distribution of GABAergic neurons and terminals in the rat CNS as revealed by GAD immunohistochemistry. In: Björklund A, Hökfelt T (eds) GABA and neuropeptides in the CNS. Handbook of chemical neuroanatomy, Vol 4. Elsevier/North-Holland, Amsterdam, pp 436–608
Murashima YL, Kato T (1985) Glutamate decarboxylase activities in single vertebrate neurons. J Neurochem 45: 732–738
Oertel WH, Schmechel DE, Tappaz ML, Kopin IJ (1981) Production of a specific antiserum to rat brain glutamic acid decarboxylase by injection of an antigen-antibody complex. Neuroscience 6: 2689–2700
Osborne NN, Beaton DW (1986) Direct histochemical localisation of 5,7-dihydroxytryptamine and the uptake of serotonin by a subpopulation of GABA neurones in the rabbit retina. Brain Res 382: 158–162
Ottersen OP, Storm-Mathisen J (1984a) Glutamate-and GABA-containing neurons in the mouse and rat brain, as demonstrated with a new immunocytochemical technique. J Comp Neurol 229: 374–392
Ottersen OP, Storm-Mathisen J (1984b) Neurons containing or accumulating transmitter amino acids. In: Björklund A, Hökfelt T, Kuhar MJ (eds) Classical transmitters and transmitter receptors in the CNS, part II. Handbook of chemical neuroanatomy, Vol 3. Elsevier/North-Holland, Amsterdam, pp 141–246
Ottersen OP, Davanger S, Storm-Mathisen J (1987) Glycine-like immunoreactivity in the cerebellum of rat and Senegalese baboon, Papio papio: a comparison with the distribution of GABA-like immunoreactivity and with [3H]glycine and [3H]GABA uptake. Exp Brain Res 66: 211–221
Paxinos G, Watson C (1982) The rat brain in stereotaxic coordinates. Academic Press, New York
Phelps PE, Vaughn JE (1986) Immunocytochemical localization of choline acetyltransferase in rat ventral striatum: a light and electron microscopic study. J Neurocytol 15: 595–617
Seguela P, Geffard M, Buijs RM, Le Moal M (1984) Antibodies against γ-aminobutyric acid: specificity studies and immunocytochemical results. Proc Natl Acad Sci USA 81: 3888–3892
Sofroniew MV, Eckenstein F, Thoenen H, Cuello AC (1982) Topography of choline acetyltransferase-containing neurons in the forebrain of the rat. Neurosci Lett 33: 7–12
Somogyi P, Takagi T (1982) A note on the use of picric acid-paraformaldehyde-glutaraldehyde fixative for correlated light and electron microscopic immunocytochemistry. Neuroscience 7: 1779–1783
Takeda N, Inagaki S, Shiosaka S, Taguchi Y, Oertel WH, Tohyama M, Watanabe T, Wada H (1984) Immunohistochemical evidence for the coexistence of histidine decarboxylase-like and glutamate decarboxylase-like immunoreactivities in nerve cells of the magnocellular nucleus of the posterior hypothalamus of rats. Proc Natl Acad Sci USA 81: 7647–7650
Vaughn JE, Famiglietti EV Jr, Barber RP, Saito K, Roberts E, Ribak CE (1981) GABAergic amacrine cells in rat retina: immunocytochemical identification and synaptic connectivity. J Comp Neurol 197: 113–127
Voigt T (1986) Cholinergic amacrine cells in the rat retina. J Comp Neurol 248: 19–35
Woolf NJ, Butcher LL (1985) Cholinergic systems in the rat brain. II. Projections to the interpeduncular nucleus. Brain Res Bull 14: 63–83
Woolf NJ, Butcher LL (1986) Cholinergic systems in the rat brain. III. Projections from the pontomesencephalic tegmentum to the thalamus, tectum, basal ganglia, and basal forebrain. Brain Res Bull 16: 603–637
Woolf NJ, Eckenstein F, Butcher LL (1984) Cholinergic systems in the rat brain. I. Projections to the limbic telencephalon. Brain Res Bull 13: 751–784
Author information
Authors and Affiliations
Additional information
This paper is dedicated to Prof. Hama on the occasion of his 66th birthday
Rights and permissions
About this article
Cite this article
Kosaka, T., Tauchi, M. & Dahl, J.L. Cholinergic neurons containing GABA-like and/or glutamic acid decarboxylase-like immunoreactivities in various brain regions of the rat. Exp Brain Res 70, 605–617 (1988). https://doi.org/10.1007/BF00247609
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00247609