, Volume 89, Issue 1, pp 25–33 | Cite as

Coexistence of GABA-and choline acetyltransferase (ChAT)-like immunoreactivity in the hypoglossal nucleus of the rat

  • M. S. Davidoff
  • W. Schulze


Single and sequential double immunocytochemical techniques were applied to localize gamma-aminobutyric acid (GABA)-and choline acetyltransferase (ChAT)-like immunoreactivity (-LI) in the hypoglossal nucleus of the rat. After subsequential double staining a relatively high number of hypoglossal motor neurons showed the coexistence of both ChAT-and GABA-LI. Coexistence of both substances was also revealed in the axons of the hypoglossal nerve situated within the medulla oblongata. Cells showing only ChAT-or GABA-LI were also observed. Differences in immunostaining between the different cell groups of the hypoglossal nucleus were established.

Following axotomy of the right hypoglossal nerve, a decrease or loss of the immunoreactivity for both ChAT and GABA in the motor neurons was established until the 3rd week after the operation. The results obtained do not give evidence on the origin of the GABA-like immunoreactive material and its functional significance in the cholinergie neurons. It can be only speculated that the GABA-like material is either taken up from the intercellular space or is synthesized by the ChAT-LI nerve cells. Functionally, the importance of GABA for the synthesis of gamma-hydroxybutyrate (a novel neurotransmitter candidate) and its postsynaptic transmitter action or presynaptic regulatory action (through autoreceptors in the membrane of the nerve endings) on the release of acetylcholine (ACh) should be taken into consideration.


Choline Acetylcholine Motor Neuron Nerve Cell Nerve Ending 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Agardh E, Yeh HH, Herrmann R, Puro DG (1985) γ-Aminobutyric acid-mediated inhibition at cholinergic synapses formed by cultured retinal neurons. Brain Res 330:323–328Google Scholar
  2. Agardh E, Brunn A, Ehinger B, Storm-Mathisen J (1986) GABA immunoreactivity in the retina. Ophthalmol Vis Sci 27:674–678Google Scholar
  3. Agardh E, Ehinger B, Wu J-Y (1987) GABA and GAD-like immunoreactivity in the primate retina. Histochemistry 86:485–490Google Scholar
  4. Altschuler RA, Parakkal MH, Fex J (1983) Localization of enkephalin-like immunoreactivity in acetylcholinesterase-positive cells in the guinea-pig lateral superior olivary complex that project to the cochlea. Neuroscience 9:621–630Google Scholar
  5. Barber RP, Vaughn JE, Roberts E (1982) The cytoarchitecture of GABAergic neurons in rat spinal cord. Brain Res 238:305–328Google Scholar
  6. Basbaum AI, Glazer EJ, Oertel W (1986) Immunoreactive glutamic acid decarboxylase in the trigeminal nucleus caudalis of the cat: a light and electron-microscopic analysis. Somatosens Res 4:77–94Google Scholar
  7. Belin MF, Nanopoulos D, Didier M, Aguera M, Steinbush H, Verhofstad A, Maitre M, Pujol JF (1983) Immunocytochemical 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 275:329–339Google Scholar
  8. Bonnano G, Raiteri M (1986) GABA enhances acetylcholine release from hippocampal nerve endings through a mechanism blocked by a GABA uptake inhibitor. Neurosci Lett 70:360–363Google Scholar
  9. Bowery NG, Hill DR, Hudson AL, Price GW, Turnbull MJ, Wilkin GP (1984) Heterogeneity of mammalian GABA receptors. In: Bowery NG (ed) Actions and interactions of GABA and benzodiazepines. Raven Press, New York, pp 81–108Google Scholar
  10. Brennan MJW (1982) GABA autoreceptors are not coupled to benzodiazepine receptors in the rat cerebral cortex. J Neurochem 38:264–266Google Scholar
  11. Burnstock G (1983) Autonomic neurotransmitters and trophic factors. J Autonom Nerv Syst 7:213–217Google Scholar
  12. Celio M (1985) Most GABA-ergic neurons in cerebral cortex and hippocampus contain the calcium-binding protein parvalbumin. Acta Anat 121:247Google Scholar
  13. Chan-Palay V, Nilaver G, Palay SL, Beinfeld MC, Zimmerman EA, Wu J-Y, O'Donohue TL (1981) Chemical heterogeneity in cerebellar Purkinje cells: existence and coexistence of glutamic acid decarboxylase-like and motilin-like immunoreactivities. Proc Natl Acad Sci USA 78:7787–7791Google Scholar
  14. Chia-Sheng Ling SLM, Schmechel DE (1986) Glutamic acid decarboxylase and somatostatin immunoreactivities in rat visual cortex. J Comp Neurol 244:369–383Google Scholar
  15. Contamina P, Sáenz de Cabezón A, Parra P, Ramón y Cajal-Agüeras S, Ramo C, Pinilla MJ, Arino MP (1987) Distribution of neural elements containing GABA-like immunoreactivity in visual centres of the rabbit. Acta Anat 130:19Google Scholar
  16. Cruz L, Basbaum AI (1985) Multiple opioid peptides and the modulation of pain: Immunohistochemical analysis of dynorphin and enkephalin in the trigeminal nucleus caudalis and spinal cord of the cat. J Comp Neurol 230:331–348Google Scholar
  17. Curtis EM, Stewart MG (1986) Development of γ-aminobutyric acid immunoreactivity in chick hyperstriatum ventrale and cerebellum: light and electron microscopical observations. Dev Brain Res 30:189–199Google Scholar
  18. Dale HH, Feldberg W, Vogt M (1936) Release of acetylcholine at voluntary motor preve endings. J Physiol (Lond) 86:353–380Google Scholar
  19. Davidoff M (1973) Über die Glia im Hypoglossuskern der Ratte nach Axotomy. Z Zellforsch 141:427–442Google Scholar
  20. Davidoff MS, Irintehev AM (1986) Acetylcholinesterase activity and type C synapses in the hypoglossal, facial and spinal cord motor nuclei of rats. An electron-microscope study. Histochemistry 84:515–524Google Scholar
  21. DeGroat WC (1970) The actions of γ-aminobutyric acid on mammalian autonomic ganglia. J Pharmacol Exp Ther 172:384–386Google Scholar
  22. Desarmenien M, Feltz P, Occhipiniti G, Santangelo E, Schlihter R (1984) Coexistence of GABA a and GABA b receptors on A and C primary afferents. Br J Pharmacol 81:327–333Google Scholar
  23. Eckenstein F, Thoenen H (1982) Production of specific antisera and monoclonal antibodies to choline acetyltransferase: characterization and use for identification of cholinergic neurons. EMBO J 1:363–368Google Scholar
  24. Eckenstein F, Thoenen H (1983) Cholinergic neurons in the rat cerebral cortex demonstrated by immunohistochemical localization of choline acetyltransferase. Neurosci Lett 36:211–215Google Scholar
  25. Eugène D (1987) Fast non-cholinergic depolarizing postsynaptic potentials in neurons of rat superior cervical ganglia. Neurosci Lett 78:51–56Google Scholar
  26. Feldberg W, Gaddum JH (1934) The chemical transmitter at synapses in a sympathetic ganglion. J Physiol (Lond) 81:305–319Google Scholar
  27. Geffard M, Vieillemaringe J, Heinrich-Rock A-M, Duris P (1985) Anti-acetylcholine antibodies and first immunocytochemical application in insect brain. Neurosci Lett 57:1–6Google Scholar
  28. Hallanger AE, Wainer BH, Rye DB (1986) Colocalization of gamma-aminobutyric acid and acetylcholinesterase in rodent cortical neurons. Neuroscience 19:763–770Google Scholar
  29. Hendry SHC, Jones EG (1986) Reduction in number of immunostained GABAergic neurons in deprived-eye dominance columns of monkey area 17. Nature 320:750–753Google Scholar
  30. Hendry SHC, Jones EG, DeFelipe J, Schmechel D, Brandon C, Emson PC (1984) Neuropeptide containing neurons of the cerebral cortex are also GABAergic. Proc Natl Acad Sci USA 81:6526–6530Google Scholar
  31. Houser CR, Crawford GD, Barber RP, Salvaterra PM, Vaughn JE (1983a) Organization and morphological characteristics of cholinergic neurons: an immunocytochemical study with a monoclonal antibody to choline acetyltransferase. Brain Res 266:97–119Google Scholar
  32. Houser CR, Lee M, Vaughn JE (1983b) Immunocytochemical localization of glutamic acid decarboxylase in normal and deafferented superior colliculus. Evidence for reorganization of γ-aminobutyric acid synapses. J Neurosci 3:2030–2042Google Scholar
  33. Kaneko T, Tashiro K, Sugimoto T, Konishi A, Mizuno N (1985) Identification of thalamic neurons with vasoactive intestinal polypeptide-like immunoreactivity in the rat. Brain Res 347:390–393Google Scholar
  34. Kato E, Kuba K, Koketsu K (1978) Presynaptic inhibition by γ-aminobutyric acid in bullfrog sympathetic ganglion cells. Brain Res 153:398–402Google Scholar
  35. Katsumaru H, Murakami F, Wu J-V, Tsukaharn N (1986) Sprouting of GABAergic synapses in the red nucleus after lesions of the nucleus interpositus in the cat. J Neurosci 6:2864–2874Google Scholar
  36. Kimura H, McGeer PL, Peng J-H, McGeer EG (1981a) The central cholinergic system studied by choline acetyltransferase immunohistochemistry in the cat. J Comp Neurol 200:151–201Google Scholar
  37. Kimura H, McGeer PL, Peng JH, McGeer EG (1981b) Mapping of cholinergic system in rostral forebrain of the rodent. In: Pepeu G, Ladinsky H (eds) Cholinergic mechanisms Plenum Press, New York London, pp 695–704Google Scholar
  38. Kimura H, McGeer PL, Peng J-H (1984) Choline acetyltransferase containing neurons in the rat brain. In: Björklung A, Hökfelt T, Kuhar MJ (eds) Handbook of chemical neuroanatomy. Vol 3: Classical transmitters and transmitter receptors in the CNS, part II. Elsevier, Amsterdam, pp 51–67Google Scholar
  39. Krnjevic K (1984) Some functional consequences of GABA uptake by brain cells. Neurosci Lett 47:282–287Google Scholar
  40. Leránth C, Fehér E (1983) Synaptology and sources of vasoactive intestinal polypeptide and substance P containing axons of the rat celiac ganglion. An experimental electron microscopic immunohistochemical study. Neuroscience 10:947–958Google Scholar
  41. Levey AI, Bolam JP, Rye DB, Hallanger AE, Demuth RM, Mesulam MM, Wainer BH (1986) A light and electron microscopic procedure for sequential double antigen localization using diaminobenzidine and benzidine dihydrochloride. J Histochem Cytochem 34:1449–1457Google Scholar
  42. Lundberg JM, Hökfelt T (1986) Multiple co-existence of peptides and classical transmitters in peripheral autonomic and sensory neurons — functional and pharmacological implications. Prog Brain Res 68:241–262Google Scholar
  43. Lundberg JM, Hökfelt T, Schultzberg M, Uvuaes-Wallenstein K, Köhler C, Said ST (1979) Occurrence of vasoactive intestinal polypeptide in certain cholinergic neurons of the cat: evidence from combined immunocytochemistry and acetylcholinesterase staining. Neuroscience 4:1539–1559Google Scholar
  44. Maley B, Newton BW (1985) Immunohistochemistry of γ-aminobutyric acid in the cat nucleus tractus solitarius. Brain Res 330:364–368Google Scholar
  45. Matute C, Streit P (1986) Monoclonal antibodies demonstrating GABA-like immunoreactivity. Histochemistry 86:147–157Google Scholar
  46. Melander T, Staines WA (1986) A galanin-like peptide coexists in putative cholinergic somata of the septum-basal forebrain complex and in acetylcholinesterase-containing fibers and varicosities within the hippocampus in the owl monkey (Aotus triirgatus). Neurosci Lett 68:17–32Google Scholar
  47. 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 10:1185–1201Google Scholar
  48. Mesulam M-M, Mufson EJ, Levey AI, Wainer BH (1984) Atlas of cholinergic neurons in the forebrain and upper brainstem of the Macaque based on monoclonal choline acetyltransferase immunohistochemistry and acetylcholinesterase histochemistry. Neuroscience 12:669–686Google Scholar
  49. Miller JA, Richter JA (1986) Effects of GABAergic drugs in vivo on high-affinity choline uptake in vitro in mouse hippocampal synaptosomes. J Neurochem 47:1916–1918Google Scholar
  50. Montero VM (1986) Localization of γ-aminobutyric acid (GABA) in type 3 cells and demonstration of their source to F2 terminals in the cat lateral geniculate nucleus: a Golgi-electron microscopic GABA-immunocytochemical study. J Comp Neurol 254:228–245Google Scholar
  51. 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) Handbook of chemical neuroanatomy Vol 4: GABA and neuropeptides in the CNS part I. Elsevier, Amsterdam, pp 436–608Google Scholar
  52. Naumann RE, Wong RKS (1984) Voltage-clamp study on GABA response desensitization in single pyramidal cells from the hippocampus of adult guinea pigs. Neurosci Lett 47:289–294Google Scholar
  53. Nishimura Y, Schwartz ML, Rakic P (1986) GABA and GAD immunoreactivity of photoreceptor terminals in primate retina. Nature 320:753–756Google Scholar
  54. Oertel WH, Schmechel DE, Mugnaini E, Tappaz ML, Kopin IJ (1981a) Immunocytochemical localization of glutamate decarboxylase in rat cerebellum with a new antiserum. Neuroscience 6:2715–2735Google Scholar
  55. Oertel WH, Schmechel DE, Tappaz ML, Kopin IJ (1981b) Production of a specific antiserum to rat brain glutamic acid decarboxylase by injection of an antigen-antibody complex. Neuroscience 6:2689–2700Google Scholar
  56. Oertel WH, Mugnaini E, Schmechel DE, Tappaz M, Kopin IJ (1982) The immunocytochemical demonstration of gamma-aminobutyric acid-ergic neurons. Methods and application. In: Palay SL, Chan-Palay V (eds) Cytochemical methods in chemical neuroanatomy Liss, New York, pp 297–329Google Scholar
  57. Oertel WH, Reithmüller G, Mugnaini E, Schmechel DE, Weindl A, Gramsch C, Herz A (1983) Opioid peptide like immunoreactivity localized in GABAergic neurons of rat neostriatum and central amygdaloid nucleus. Life Sci 33 (Suppl I):73–76Google Scholar
  58. Ottersen OP, Storm-Mathisen J (1984) Glutamate-and GABA-containing neurons in the mouse and rat brain as demonstrated with a new immunocytochemical technique. J Comp Neurol 229:374–392Google Scholar
  59. Phelps P, Vaughn JE (1986) Immunocytochemical localization of choline acetyltransferase in rat ventral striatum: a light and electron microscopic study. J Neurocytol 15:595–617Google Scholar
  60. Roberts E (1984) GABA neurons in the mammalian central nervous system: model for a minimal basic unit. Neurosci Lett 47:195–200Google Scholar
  61. Roberts E, Chase TN, Tower DB (eds) (1976) GABA in nervous system function. Raven Press, New YorkGoogle Scholar
  62. Seguela P, Geffard M, Buijis RM, LeMoal M (1984) Antibodies against gamma-aminobutyric acid: specificity studies and immunocytochemical results. Proc Natl Acad Sci USA 81:3888–3892Google Scholar
  63. Senba E, Daddona PE, Watanabe T, Wu J-Y, Nagy JI (1985) Coexistence of adenosine deaminase, histidine decarboxylase and glutamate decarboxylase in hypothalamic neurons of the rat. J Neuroscience 5:3393–3402Google Scholar
  64. Somogyi P, Hodgson AJ (1985) Antisera to γ-aminobutyric acid. III. Demonstration of GABA in Golgi-impregnated neurons and in conventional electron microscopic sections of the cat striate cortex. J Histochem Cytochem 33:249–257Google Scholar
  65. Somogyi P, Takagi H (1982) A note on the use of picric acid-paraformaldehyde-glutaraldehyde fixative for correlated light and electron microscopic immunocytochemistry. Neuroscience 7:1779–1783Google Scholar
  66. Somogyi P, Hodgson AJ, Smith AD, Nunzi MG, Gorio A, Wu HY (1984) Different populations of GABAergic neurons in the visual cortex and hippocampus of cat contain somatostatinor cholecystokinin-immunoreactive material. J Neurosci 4:2590–2603Google Scholar
  67. Somogyi P, Hodgson AJ, Chubb JW, Penke B, Erdei A (1985) Antisera to γ-aminobutyric acid. II. Immunocytochemical application to the central nervous system. J Histochem Cytochem 33:240–248Google Scholar
  68. Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG (1970) The unlabeled antibody method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horse radish peroxidase-antiperoxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333Google Scholar
  69. Storm-Mathisen J, Leknes AK, Bore AT, Vaaland JL, Edminson P, Haug FMS, Ottersen OP (1983) First visualization of glutamate and GABA in neurones by immunocytochemistry. Nature 301:517–520Google Scholar
  70. Takeda N, Inagaki S, Shiosaka T, Taguchi Y, Oertel W, Tohyama M, Watanabe T, Wada H (1984) Immunohistochemical evidence for the coexistence of GABA and histidine decarboxylase-like immunoreactivities in nerve cells of the magnocellular nucleus of the posterior hypothalamus of the rat. Proc Natl Acad Sci USA 81:7647–7650Google Scholar
  71. Tunnicliff G, Ngo TT (1986) Regulation of γ-aminobutyric acid synthesis in the vertebrate nervous system. Neurochem Int 8:287–297Google Scholar
  72. Verburg-Van Kemenade BML, Jenks BG, Driessen AGJ (1986) GABA and dopamine act directly on melanophores of Xenopus to inhibit MSH secretion. Brain Res Bull 17:697–704Google Scholar
  73. Vincent S, Hökfelt T, Christensson I, Terenius L (1982) Immunohistochemical evidence for a dynorphin immunoreactive striatonigral pathway. Eur J Pharmacol 85:251–252Google Scholar
  74. Vincent SR, Satoch K, Fibiger HC, Panula P, Armstrong DM (1984) Peptides in the ascending cholinergic reticular system. Abstracts of the VIIth International Congress of Histochemistry and Cytochemistry, Helsinki, August 5–11, p 466Google Scholar
  75. Vuillez P, Pérez SC, Stoeckel ME (1987) Colocalization of GABA and tyrosine hydroxylase immunoreactivities in the axons innervaling the neurointermediate lobe of the rat pituitary: an ultrastructural immunogold study. Neurosci Lett 79:53–58Google Scholar
  76. Wainer BH, Bolam JP, Freund TF, Henderson Z, Totterdell S, Smith AD (1984a) Cholinergic synapses in the rat brain: a correlated light and electron microscopic immunohistochemical study employing a monoclonal antibody against choline acetyltransferase. Brain Res 308:69–76Google Scholar
  77. Wainer BH, Levey I, Mufson EJ, Mesulam MM (1984b) Cholinergic systems in mammalian brain identified with antibodies against choline acetyltransferase. Neurochem Int 6:163–182Google Scholar
  78. Wassef M, Simons J, Tappaz ML, Sotelo C (1986) Non-Purkinje cell GABAergic innervation of the deep cerebellar nuclei: A quantitative immunocytochemical study in C57BL and in Purkinje cell degeneration mutant mice. Brain Res 399:125–135Google Scholar
  79. Watt CB, Su YT, Lam DM-K (1984) Interactions between enkephalin and GABA in avian retina. Nature 311:761–763Google Scholar
  80. Weissman-Nanopoulos D, Belin MF, Mandel P, Maitre M (1984) Immunocytochemical evidence for the presence of enzymes synthesizing GABA and GHB in the same neuron. Neurochem Int 6:333–338Google Scholar
  81. Wolff JR, Böttcher H, Zetzsche T, Oertel WH, Chronwall BM (1984) Development of GABAergic neurons in rat visual cortex as identified by glutamate decarboxylase-like immunoreactivity. Neurosci Lett 47:207–212Google Scholar
  82. Wolff JR, Joo F, Kasa P, Storm-Mathisen J, Toldi J, Balcar VJ (1986) Presence of neurons with GABA-like immunoreactivity in the superior cervical ganglion of the rat. Neurosci Lett 71:157–162Google Scholar
  83. Wu J-Y (1976) Purification and properties of l-Glutamate decarboxylase (GAD) and GABA-aminotransferase (GABA-T). In: Roberts E, Chase T, Tower D (eds) GABA in nervous system function. Raven Press, New York, pp 7–55Google Scholar
  84. Wu J-Y (1983) Preparation of glutamic acid decarboxylase as immunogen for immunocytochemistry. In: Cuello AC (ed) IBRO handbook series: Methods in the neurosciences. Wiley, London New York, pp 159–191Google Scholar
  85. Wu J-Y, Lin C-T, Brandon C, Chan T-S, Moehler H, Richards JG (1982) Regulation and immunocytochemical characterization of glutamic acid decarboxylase. In: Palay S, Chan-Palay V (eds) Cytochemical methods in neuroanatomy. Alan R Liss, New York, pp 279–296Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • M. S. Davidoff
    • 1
  • W. Schulze
    • 2
  1. 1.Regeneration Research LaboratoryBulgarian Academy of SciencesSofiaBulgaria
  2. 2.Institute of AnatomyUniversity of HamburgHamburg 20Germany

Personalised recommendations