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Immunocytochemical localization of choline acetyltransferase in rat ventral striatum: a light and electron microscopic study

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Journal of Neurocytology

Summary

The ventral striatum, previously defined as including the nucleus accumbens, substriatal grey, olfactory tubercle and striatal cell bridges has been examined in an immunocytochemical study with monoclonal antibodies to choline acetyltransferase (ChAT) in order to identify putative cholinergic neurons and synaptic junctions within the region. Light microscopy revealed ChAT-positive neurons with similar morphological characteristics in all divisions of ventral striatum. The somata of immunoreactive neurons were round or elongated in shape, approximately 10 × 21 μm in size and had two to four dendrites that coursed long distances and occasionally branched. Electron microscopy of ChAT-positive neurons in substriatal grey initially studied by light microscopy revealed that unlabelled boutons occasionally formed synapses with immunoreactive somata and proximal dendrites, but were more numerous along distal dendrites. Light microscopy demonstrated that ventral striatal neuropil contained numerous ChAT-positive fibres and punctate structures that varied in concentration from moderate to very dense. The lateral border of the substriatal grey and the area within, and adjacent to, all islands of Calleja exhibited the most dense ChAT-positive punctate staining. Additionally, the medial portion of nucleus accumbens was more densely ChAT-positive than the lateral, and the olfactory tubercle displayed laminar variations of immunoreaction product. Counterstained immunocytochemical specimens demonstrated that some areas of dense ChAT-positive punctate staining were associated with clusters of ChAT-negative, medium-sized neurons. Furthermore, electron microscopic observations of substriatal grey revealed that ChAT-positive dense regions were associated with numerous immunoreactive boutons, some of which established synapses with unlabelled somata, dendritic shafts and spines. These results suggest that the densely ChAT-positive neuropil areas within ventral striatum receive more cholinergic innervation than the more lightly stained neuropil areas. There are numerous similarities in the morphological characteristics of ChAT-positive neurons and synapses observed in ventral striatum when compared with those previously described in dorsal striatum. However, some differences were observed, such as smaller somal sizes in ventral, as contrasted with dorsal striatum, and a substantial variation in ChAT-positive fibre and punctate neuropil staining seen within the ventral but not the dorsal striatum. Such differences suggest that the ventral striatum may exhibit greater heterogeneity of cholinergic function than the dorsal striatum.

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References

  • Armstrong, D. M., Saper, C. B., Levey, A. I., Wainer, B. H. &Terry, R. D. (1983) Distribution of cholinergic neurons in rat brain: demonstrated by the immunocytochemical localization of choline acetyltransferase.Journal of Comparative Neurology 216, 53–68.

    Google Scholar 

  • Björklund, A. &Lindvall, O. (1984) Dopaminecontaining systems in the CNS. InHandbook of Chemical Neuroanatomy, Vol. 2,Classical Transmitters in the CNS, Part I (edited byBjörklund, A. &Hökfelt, T.), pp. 55–122. Amsterdam: Elsevier Biomedical Press.

    Google Scholar 

  • Bolam, J. P., Wainer, B. H. &Smith, A. D. (1984) Characterization of cholinergic neurons in the rat neostriatum. A combination of choline acetyltransferase immunocytochemistry, Golgi-impregnation and electron microscopy.Neuroscience 12, 711–18.

    Google Scholar 

  • Brennan, L. A., Phelps, P. E. &Barber, R. P. (1986) An improved method of toluidine blue counterstaining of neurons in immunocytochemical preparations.Stain Technology 61 (in press).

  • Butcher, L. L. &Woolf, N. J. (1982) Cholinergic and serotonergic systems in the brain and spinal cord: anatomic organization, role in intercellular communication processes, and interactive mechanisms. InChemical Transmission in the Brain, Progress in Brain Research, Vol. 55 (edited byBuijs, R. M., Pevet, P. &Swaab, D. F.), pp. 3–40. Amsterdam: Elsevier Biomedical Press.

    Google Scholar 

  • Chang, H. T. &Kitai, S. T. (1985) Projection neurons of the nucleus accumbens: An intracellular labeling study.Brain Research 347, 112–16.

    Google Scholar 

  • Chang, H. T., Wilson, C. J. &Kitai, S. T. (1982) A Golgi study of rat neostriatal neurons: light microscopic analysis.Journal of Comparative Neurology 208, 107–26.

    Google Scholar 

  • Crawford, G. D., Correa, L. &Salvaterra, P. M. (1982) Interaction of monoclonal antibodies with mammalian choline acetyltransferase.Proceedings of the National Academy of Sciences USA 79, 7031–5.

    Google Scholar 

  • DiFiglla, M., Pasik, T. &Pasik, P. (1980) Ultrastructure of Golgi-impregnated and gold-toned spiny and aspiny neurons in the monkey striatum.Journal of Neurocytology 9, 471–92.

    Google Scholar 

  • Dimova, R., Vuillet, J. &Seïte, R. (1980) Study of the rat neostriatum using a combined Golgi-electron microscope technique and serial sections.Neuroscience 5, 1581–96.

    Google Scholar 

  • Domesick, V. B. (1981) Further observations on the anatomy of nucleus accumbens and caudatoputamen in the rat: similarities and contrasts. InThe Neurobiology of the Nucleus Accumbens (edited byChronister, R. B. &De France, J. F.), pp. 7–39. Brunswick: Haer Institute.

    Google Scholar 

  • Fairén, A., Peters, A. &Saldanha, J. (1977) A new procedure for examining Golgi impregnated neurons by light and electron microscopy.Journal of Neurocytology 6, 311–37.

    Google Scholar 

  • Fallon, J. H., Riley, J. N., Sipe, J. C. &Moore, R. Y. (1978) The islands of Calleja: organization and connections.Journal of Comparative Neurology 181, 375–96.

    Google Scholar 

  • Fonnum, F., Walaas, I. &Iversen, E. (1977) Localization of GABAergic, cholinergic and aminergic structures in the mesolimbic system.Journal of Neurochemistry 29, 221–30.

    Google Scholar 

  • Fox, C. A., Andrade, A. N., Hillman, D. E. &Schwyn, R. C. (1971/1972) The spiny neurons in the primate striatum: a Golgi and electron microscopic study.Journal für Hirnforschung 13, 181–201.

    Google Scholar 

  • Graybiel, A. M. &Ragsdale, C. W. Jr. (1979) Fiber connections of the basal ganglia. InDevelopment and Chemical Specificity of Neurons (edited byCuenod, M., Kreutzberg, G. W. &Bloom, F. E.), pp. 239–83. Amsterdam: Elsevier Biomedical Press.

    Google Scholar 

  • Gurdjian, E. S. (1927) The corpus striatum of the rat.Journal of Comparative Neurology 45, 249–81.

    Google Scholar 

  • Haber, S. N., Groenewegen, H. J., Grove, E. A. &Nauta, W. J. H. (1985) Efferent connections of the ventral pallidum: evidence of a dual striato pallidofugal pathway.Journal of Comparative Neurology 235, 322–35.

    Google Scholar 

  • Hedreen, J. C. (1981) Neurons of the nucleus accumbens and other striatal regions in rats. InThe Neurobiology of the Nucleus Accumbens (edited byChronister, R. B. &De France, J. F.), pp. 82–96. Brunswick: Haer Institute.

    Google Scholar 

  • Heimer, L. (1978) The olfactory cortex and the ventral striatum. InThe Continuing Evolution of the Limbic System Concept (edited byLivingston, K. E. &Hornykiewicz, O.), pp. 95–187. New York: Plenum Press.

    Google Scholar 

  • Heimer, L. &Wilson, R. D. (1975) The subcortical projections of the allocortex: similarities in the neural associations of the hippocampus, the piriform cortex, and the neocortex. InGolgi Centennial Symposium, Proceedings (edited bySantini, M.), pp. 177–93. New York: Raven Press.

    Google Scholar 

  • Herkenham, M., Moon Edley, S. &Stuart, J. (1984) Cell clusters in the nucleus accumbens of the rat, and the mosaic relationship of opiate receptors, acetylcholinesterase and subcortical afferent terminations.Neuroscience 11, 561–93.

    Google Scholar 

  • Hoover, D. B., Muth, E. A. &Jacobowitz, D. M. (1978) A mapping of the distribution of acetylcholine, choline acetyltransferase and acetylcholinesterase in discrete areas of rat brain.Brain Research 153, 295–306.

    Google Scholar 

  • Houser, C. R., Barber, R. P., Crawford, G. D., Matthews, D. A., Phelps, P. E., Salvaterra, P. M. &Vaughn, J. E. (1984) Species-specific second antibodies reduce spurious staining in immunocytochemistry.Journal of Histochemistry and Cytochemistry 32, 395–402.

    Google Scholar 

  • Houser, C. R., Crawford, C. D., Barber, R. P., Salv/Terra, P. M. &Vaughn, J. E. (1983) Organization and morphological characteristics of cholinergic neurons: an immunocytochemical study with a monoclonal antibody to choline acetyltransferase.Brain Research 266, 97–119.

    Google Scholar 

  • Ingham, C. A., Bolam, J. P., Wainer, B. H. &Smith, A. D. (1985) A correlated light and electron microscopic study of identified cholinergic basal forebrain neurons, that project to the cortex in the rat.Journal of Comparative Neurology 239, 176–92.

    Google Scholar 

  • Kemp, J. M. &Powell, T. P. S. (1970) The cortico-striate projection in the monkey.Brain 93, 525–46.

    Google Scholar 

  • Kemp, J. M. &Powell, T. P. S. (1971) The synaptic organization of the caudate nucleus.Philosophical Transactions of the Royal Society of London, Series B 262, 403–12.

    Google Scholar 

  • Kimura, H., McGeer, P. L. &Peng, J.-H. (1984) Choline acetyltransferase-containing neurons in the rat brain. InHandbook of Chemical Neuroanatomy, Vol. 3,Classical Transmitters and Transmitter Receptors in the CNS, Part II (edited byBjörklund, A., Hökfelt, T. &Kuhar, M. J.), pp. 51–67. Amsterdam: Elsevier.

    Google Scholar 

  • Kimura, H., McGeer, P. L., Peng, J.-H. &McGeer, E. G. (1981) The central cholinergic system studied by choline acetyltransferase immunohistochemistry in the cat.Journal of Comparative Neurology 200, 151–201.

    Google Scholar 

  • McGeer, P. L., McGeer, E. G., Fibiger, H. C. &Wickson, V. (1971) Neostriatal choline acetylase and cholinesterase following selective brain lesions.Brain Research 35, 308–14.

    Google Scholar 

  • Mesulam, M.-M., Mufson, E. J., Levey, A. I. &Wainer, B. H. (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, 669–86.

    Google Scholar 

  • Miller, M. M., Goto, R. &Clark, S. D. (1982) Structural characterization of developmentally expressed antigenic markers on chicken erythrocytes using monoclonal antibodies.Developmental Biology 94, 400–14.

    Google Scholar 

  • Millhouse, O. E. &Heimer, L. (1984) Cell configurations in the olfactory tubercle of the rat.Journal of Comparative Neurology 228, 571–97.

    Google Scholar 

  • Mugnaini, E. &Oertel, W. H. (1985) An atlas of the distribution of GABAergic neurons and terminals in the rat CNS as revealed by GAD immunohistochemistry. InHandbook of Chemical Neuroanatomy, Vol. 4,GABA and Neuropeptides in the CNS, Part I (edited byBjörklund, A. &Hökfelt, T.), pp. 436–622. Amsterdam: Elsevier.

    Google Scholar 

  • Nauta, W. J. H., Smith, G. P., Faull, R. L. M. &Domesick, V. B. (1978) Efferent connections and nigral afferents of the nucleus accumbens septi in the rat.Neuroscience 3, 385–401.

    Google Scholar 

  • Newman, R. &Winans, S. S. (1980a) An experimental study of the ventral striatum of the golden hamster. I. Neuronal connections of the nucleus accumbens.Journal of Comparative Neurology 191, 167–92.

    Google Scholar 

  • Newman, R. &Winans, S. S. (1980b) An experimental study of the ventral striatum of the golden hamster. II. Neuronal connections of the olfactory tubercule.Journal of Comparative Neurology 191, 193–212.

    Google Scholar 

  • Oertel, W. H. &Mugnaini, E. (1984) Immunocytochemical studies of GABAergic neurons in rat basal ganglia and their relations to other neuronal systems.Neuroscience Letters 47, 233–8.

    Google Scholar 

  • Ordronneau, P., Lindström, P. B.-M. &Petrusz, P. (1981) Four unlabeled antibody bridge techniques: a comparison.Journal of Histochemistry and Cytochemistry 29, 1397–404.

    Google Scholar 

  • Palkovits, M., Saavedra, J. M., Kobayashi, R. M. &Brownstein, M. (1974) Choline acetyltransferase content of limbic nuclei of the rat.Brain Research 79, 443–50.

    Google Scholar 

  • Paxinos, G. &Watson, C. (1982)The Rat Brain in Stereotaxic Coordinates. New York: Academic Press.

    Google Scholar 

  • Phelps, P. E., Houser, C. R. &Vaughn, J. E. (1985) Immunocytochemical localization of choline acetyltransferase within the rat neostriatum: a correlated light and electron microscopic study of cholinergic neurons and synapses.Journal of Comparative Neurology 238, 286–307.

    Google Scholar 

  • Ribak, C. E. &Fallon, J. H. (1982) The island of Calleja complex of rat basal forbrain. I. Light and electron microscopic observations.Journal of Comparative Neurology 205, 207–18.

    Google Scholar 

  • Sofroniew, M. V., Eckenstein, F., Thoenen, H. &Cuello, A. C. (1982) Topography of choline acetyltransferase-containing neurons in the forebrain of the rat.Neuroscience Letters 33, 7–12.

    Google Scholar 

  • Swanson, L. W. &Cowan, W. M. (1975) A note on the connections and development of the nucleus accumbens.Brain Research 92, 324–30.

    Google Scholar 

  • Switzer, R. C., III, Hill, J. &Heimer, L. (1982) The globus pallidus and its rostroventral extension into the olfactory tubercle of the rat: a cyto- and chemoarchitectural study.Neuroscience 7, 1891–904.

    Google Scholar 

  • Wainer, B. H., Bolam, J. P., Freund, T. F., Henderson, Z., Totterdell, S. &Smith, A. D. (1984) Cholinergic synapses in the rat brain: a correlated light and electron microscopic immunohistochemical study employing a monoclonal antibody against choline acetyltransferase.Brain Research 308, 69–76.

    Google Scholar 

  • Walaas, I. &Fonnum, F. (1979a) The distribution and origin of glutamate decarboxylase and choline acetyltransferase in ventral pallidum and other basal forebrain regions.Brain Research 177, 325–36.

    Google Scholar 

  • Walaas, I. &Fonnum, F. (1979b) The effects of surgical and chemical lesions on neurotransmitter candidates in the nucleus accumbens of the rat.Neuroscience 4, 209–16.

    Google Scholar 

  • Winer, B. J. (1971)Statistical Principles in Experimental Design, pp. 210–18. New York: McGraw-Hill.

    Google Scholar 

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  1. Division of Neurosciences, Beckman Research Institute of the City of Hope, 91010, Duarte, California, USA

    Patricia E. Phelps & James E. Vaughn

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  1. Patricia E. Phelps
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  2. James E. Vaughn
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Phelps, P.E., Vaughn, J.E. Immunocytochemical localization of choline acetyltransferase in rat ventral striatum: a light and electron microscopic study. J Neurocytol 15, 595–617 (1986). https://doi.org/10.1007/BF01611860

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  • DOI: https://doi.org/10.1007/BF01611860

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