Advertisement

Journal of Neurocytology

, Volume 14, Issue 6, pp 909–919 | Cite as

Choline acetyltransferase immunoreactivity is localized to four types of synapses in the rat interpeduncular nucleus

  • Nicholas J. Lenn
  • Csaba Leranth
  • Laszlo Zaborszky
Article

Summary

The cholinergic synapses of the rat interpeduncular nucleus (IPN) were demonstrated by immunostaining that utilized a monoclonal antibody directed against choline acetyltransferase. The rostral, central, intermediate and lateral subnuclei of the IPN each contained a single type of immunoreactive terminal. Immunoreactivity was localized to synaptic vesicle membranes, inner and outer mitochondrial membranes, the cytoplasmic surface of plasma membranes (especially at the contact zones), and longitudinal microtubules in preterminal portions of axons. Terminals were identified by comparison to previous studies of the synaptic organization of the IPN. In the rostral subnucleus, the immunoreactive terminals were characterized by their content of spherical vesicles, 45 nm in diameter, intermixed with moderate numbers of dense-cored vesicles, 75–100 nm in diameter. These terminals formed asymmetrical contacts. They correspond to the more numerous of the two types of axodendritic terminals described in this subnucleus, i.e. those which degenerate after lesions of the habenula. The moderate number of immunoreactive terminals in the lateral subnucleus contained pleomorphic vesicles, 30–45 nm in diameter. Up to three of these formed symmetrical contacts with individual dendrites, which ranged in diameter from 0.35 to 0.55μm. The other types of axodendritic terminal in this subnucleus, which often contacted the same dendrites, were unstained. These latter terminals have been interpreted as being those which contain substance P. The immunoreactive terminals in the central subnucleus consisted of moderate numbers of S terminals. These contained spherical vesicles, 40–60 nm in diameter, and formed markedly asymmetricalen passant contacts with small dendritic processes or spines. The immunoreactive terminals in the intermediate subnucleus had the same presynaptic and contact morphology. Many were clearly crest synapses. The remainder appeared to be such, but seen only partially within the plane of section. In the intermediate subnucleus there were up to several hundred immunostained terminals per grid square in some sections.

These findings are consistent with the existence of a dense cholinergic projection to the IPN. The crest and S synapses, both of which degenerate after lesions of the habenular region, are shown to be cholinergic as previously suggested. Additionally, demonstration of the cholinergic innervation of other subnuclei of the IPN increases understanding of the relation of cholinergic to other transmitters localized to various portions of this nucleus.

Keywords

Moderate Number Choline Acetyltransferase Spherical Vesicle Synaptic Vesicle Membrane Individual Dendrite 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Artymyshyn, R. &Murray, M. (1985) Substance P in the interpeduncular nucleus of the rat: normal distribution and the effects of deafferentation.Journal of Comparative Neurology 231, 78–90.PubMedGoogle Scholar
  2. Eckenstein, F., Barde, Y-A. &Thoenen, H. (1981) Production of specific antibodies to choline acetyltransferase purified from pig brain.Neuroscience 6, 993–1000.PubMedGoogle Scholar
  3. Eckenstein, F., &Thoenen, H. (1982) Production of specific antisera and monoclonal. Antibodies to choline acetyltransferase: characterization and use for identification of cholinergic neurons.EMBO Journal 1, 363–8.PubMedGoogle Scholar
  4. Geary, W. A., II &Wooten, G. F. (1983) Quantitative film autoradiography of opiate agonist and antagonist binding in rat brain.Journal of Pharmacology and Experimental Therapeutics 225, 234–40.PubMedGoogle Scholar
  5. Gottesfeld, Z. &Jacobowitz, D. M. (1978) Cholinergic projection of the diagonal band to the interpeduncular nucleus of the rat brain.Brain Research 156, 329–32.PubMedGoogle Scholar
  6. Hamill, G. S. &Fass, B. (1984) Differential distribution of diagonal band afferents to subnuclei of the interpeduncular nucleus in rats.Neuroscience Letters 48, 43–8.PubMedGoogle Scholar
  7. Hamill, G. S. &Lenn, N. J. (1984) The subnuclear organization of the rat interpeduncular nucleus: a light and electron microscopic study.Journal of Comparative Neurology 222, 396–408.PubMedGoogle Scholar
  8. Hamill, G. S., Olschowka, J. A., Lenn, N. J. &Jacobowitz, D. M. (1984) The subnuclear distribution of substance P, cholecystokinin, vasoactive intestinal peptide, somatostatin, leu-enkephalin, dopamine-β-hydroxylase and serotonin in the rat interpeduncular nucleus.Journal of Comparative Neurology 226, 580–96.PubMedGoogle Scholar
  9. Hattori, T., McGeer, E. G., Singh, V. K. &McGeer, P. L. (1977) Cholinergic synapse of the interpeduncular nucleus.Experimental Neurology 55, 666–79.PubMedGoogle Scholar
  10. Hemmendinger, L. M. &Moore, R. Y. (1984) Interpeduncular nucleus organization in the rat: cytoarchitecture and histochemical analysis.Brain Research Bulletin 13, 163–79.PubMedGoogle Scholar
  11. Herkenham, M. &Nauta, W. J. H. (1979) Efferent connections of the habenular nuclei in the rat.Journal of Comparative Neurology 187, 19–48.PubMedGoogle Scholar
  12. Herkenham, M. &Pert, C. B. (1980) In vitro autoradiography of opiate receptors in rat brain suggests loci of ‘opiatergic’ pathways.Proceedings of the National Academy of Sciences, USA 77, 5532–6.Google Scholar
  13. Houser, C. R., Crawford, G. D., Barber, R. P., Salvaterra, 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.PubMedGoogle Scholar
  14. Hsu, S-M., Raine, L. &Fanger, H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures.Journal of Histochemistry and Cytochemistry 29, 577–80.PubMedGoogle Scholar
  15. Kapadia, S. E. &De Lanerolle, N. C. (1984) The ultrastructure and organization of methionine-enkephalin immunoreactive profiles in the interpeduncular nucleus of the cat.Journal of Comparative Neurology 229, 48–65.PubMedGoogle Scholar
  16. Kataoka, K., Nakamura, Y. &Hassler, R. (1973) Habenulo-interpeduncular tract: a possible cholinergic neuron in rat brain.Brain Research 62, 264–7.PubMedGoogle Scholar
  17. Kuhar, M. J., Dehaven, R. N., Yamamura, H. I., Rommel-Spacher, H., Simon, J. R. (1975) Further evidence for cholinergic habenulo-interpeduncular neurons: pharmacologie and functional characteristics.Brain Research 97, 265–75.PubMedGoogle Scholar
  18. Lake, N. (1973) Studies of the habenulo-interpeduncular pathway in cats.Experimental Neurology 41, 113–32.PubMedGoogle Scholar
  19. Lenn, N. J. (1976) Synapses in the interpeduncular nucleus: electron microscopy of normal and habenula lesioned rats.Journal of Comparative Neurology 166, 73–100.Google Scholar
  20. Lenn, N. J. &Bayer, S. (1985) Birthdays of neurons in subnuclei of the rat interpeduncular nucleus.Society For Neuroscience Abstracts 12, 392.Google Scholar
  21. Lenn, N. J. &Hamill, G. S. (1984) A proposed nomenclature for the subnuclei of the interpeduncular nucleus.Brain Research Bulletin 13, 203–4.PubMedGoogle Scholar
  22. Lenn, N. J. &Whitmore, L. (1984) Horseradish peroxidase filled single axons in the rat interpeduncular nucleus.Society for Neuroscience Abstracts 11, 334.Google Scholar
  23. Lenn, N. J., Wong, V. &Hamill, G. S. (1983) Left-right pairing at the crest synapses of rat interpeduncular nucleus.Neuroscience 9, 383–9.PubMedGoogle Scholar
  24. Leranth, Cs., Brownstein, M., Zaborszky, L., Jaranyi, Zs. &Palkovits, M. (1975) Morphological and biochemical changes in the rat interpeduncular nucleus following the transection of the habenulo-interpeduncular tract.Brain Research 99, 124–8.PubMedGoogle Scholar
  25. Leranth, Cs. &Feher, E. (1983) Synaptology and sources of vasoactive intestinal polypeptide and substance P containing axons of the cat celiac ganglion. An experimental electron microscopic immunohistochemical study.Neuroscience 10, 947–58.PubMedGoogle Scholar
  26. McGeer, E. G., Scherer-Singler, U. &Singh, E. A. (1979) Confirmatory data on habenular projections.Brain Research 168, 375–6.PubMedGoogle Scholar
  27. Murray, M., Zimmer, J. &Raisman, G. (1979) Quantitative electron microscopic evidence for reinnervation in the adult rat interpeduncular nucleus after lesions of the fasciculus retroflexus.Journal of Comparative Neurology 187, 447–68.PubMedGoogle Scholar
  28. 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.PubMedGoogle Scholar
  29. Rossier, J. (1981) Serum monospecificity: a prerequisite for reliable immunohistochemical localization of neuronal markers including choline acetyltransferase.Neuroscience 6, 989–91.PubMedGoogle Scholar
  30. Rotter, A. &Jacobowitz, D. M. (1984) Localization of substance P, acetylcholinesterase, muscarinic receptors and alpha-bungarotoxin binding sites in the rat interpeduncular nucleus.Brain Research Bulletin 12, 83–94.PubMedGoogle Scholar
  31. Sastry, B. R. (1978) Effects of substance P, acetylcholine and stimulation of habenula on rat interpeduncular neural activity.Brain Research 144, 404–10.PubMedGoogle Scholar
  32. Somogyi, P. &Takagi, H. (1982) A note on the use of picric acid-paraformalde-hydeglutaraldehyde fixative for correlated light and electron microscopic immunocytochemistry.Neuroscience 7, 1779–83.PubMedGoogle Scholar
  33. Sorimachi, M. &Kataoka, K. (1974) Choline uptake by nerve terminals: a sensitive and a specific marker of cholinergic innervation.Brain Research 72, 350–3.PubMedGoogle Scholar
  34. Spurr, A. R. (1969) A low-viscosity epoxy resin embedding medium for electron microscopy.Journal of Ultrastructure Research 26, 31–4.PubMedGoogle Scholar
  35. Stofer, W. D. (1980) The cytoarchitecture and efferent projections of the interpeduncular complex in the cat. Phd Thesis, Department of Anatomy, University of Virginia.Google Scholar
  36. Takagi, M. (1984) Actions of cholinergic drugs on cells in the interpeduncular nucleus.Experimental Neurology 84, 358–63.PubMedGoogle Scholar
  37. 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.PubMedGoogle Scholar

Copyright information

© Chapman and Hall Ltd. 1985

Authors and Affiliations

  • Nicholas J. Lenn
    • 1
    • 2
    • 3
  • Csaba Leranth
    • 4
  • Laszlo Zaborszky
    • 1
    • 3
  1. 1.Department of NeurologyUniversity of VirginiaCharlottesvilleUSA
  2. 2.Department of PediatricsUniversity of VirginiaCharlottesvilleUSA
  3. 3.Clinical Neuroscience Research CenterUniversity of VirginiaCharlottesvilleUSA
  4. 4.Section of Neuroanatomy and Department of Obstetrics and GynecologyYale UniversityNew HavenUSA

Personalised recommendations