Cell Biology of the Forebrain Cholinergic Neurons: Effects of NGF, Triiodothyronine and Gangliosides

  • F. Hefti


Alzheimer’s disease is associated with a selective loss of cholinergic neurons located in the basal forebrain. Even though other neuronal systems are also partly affected, the loss of cholinergic neurons is regarded by most investigators as being the principal factor responsible for the memory loss that is characteristic of Alzheimer’s disease (Bartus et al., 1982; Coyle et al., 1983; Davies, 1985). In attempting to find the cause of and treatment for the disease, I decided to study the cell biology of cholinergic neurons of the forebrain and to characterise their requirements for survival and maintenance of function. A culture system was developed in which cholinergic neurons from rat brains are grown and studied under controlled conditions and are easily accessible for observation. Using these cultures, the effects of other cell types, growth factors, hormones and drugs on survival, growth and differentiation of cholinergic cells are investigated. These studies will lead to a characterisation of the conditions required by cholinergic neurons to survive and maintain their function in vitro. Conditions and compounds found to affect cholinergic neurons in vitro will later be assessed in living animals with specific lesions of the cholinergic systems. The in vitro studies first focussed on the effects of NGF and thyroxine, which were both reported to influence cholinergic neurons, and on gangliosides, which were claimed to promote neuronal survival and regeneration.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Agnati, L. F., Fuxe, K., Calza, L., Benefanti, F., Cavicchioli, L., Toffano, G., and Goldstein, M. (1983). Gangliosides increase the survival of lesioned nigral dopamine neurons and favour the recovery of dopaminergic synaptic function in striatum of rats by collateral sprouting. Acta Physiol Scand., 119, 347–63.CrossRefPubMedGoogle Scholar
  2. Ando, S. (1983). Gangliosides in the nervous system. Neurochem. Int., 5, 507–37.CrossRefPubMedGoogle Scholar
  3. Bartus, R. T., Dean, R. L., Beer, B., and Lippa, A. S. (1982). The cholinergic hypothesis of geriatric memory dysfunction. Science, 217, 408–17.CrossRefPubMedGoogle Scholar
  4. Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248–54.CrossRefPubMedGoogle Scholar
  5. Byrne, M. C., Ledeen, F. J., Roisen, F. J., York, G., and Scalafani, J. R. (1983). Ganglioside-induced neuritogenesis: verification that gangliosides are the active agents, and comparison of molecular species. J. Neurochem., 41, 1214–22.CrossRefPubMedGoogle Scholar
  6. Ceccarelli, B., Aporti, F., and Finesso, M. (1976). Effects of brain gangliosides on functional recovery in experimental regeneration and reinnervation. Adv. Exp. Med. Biol., 71, 275–93.CrossRefPubMedGoogle Scholar
  7. Cheung, E. N. (1985). Thyroid hormone action: determination of hormone-receptor interaction using structural analogs and molecular modeling. Trends Pharmacol. Sci., Jan. 1985, 31–4.Google Scholar
  8. Coyle, J. T., Price, D. L., and DeLong, M. R. (1983). Alzheimer’s disease: a disease of cortical cholinergic innervation. Science, 219, 1184–9.CrossRefPubMedGoogle Scholar
  9. Crutcher, K. A. (1982). Development of the rat septohippocampal projection: a retrograde fluorescent tracer study. Dev. Brain Res., 3, 145–50.CrossRefGoogle Scholar
  10. Crutcher, K. A., and Collins, F. (1982). In vitro evidence for two distinct hippocampal growth factors: basis of neuronal plasticity? Science, 217, 67–70.CrossRefPubMedGoogle Scholar
  11. Crutcher, K. A., and Davis, J. N. (1981). Sympathetic noradrenergic sprouting in response to central cholinergic denervations. Trends Neurosci., 4, 70–2.CrossRefGoogle Scholar
  12. Crutcher, K. A., Brothers, L., and Davis, J. N. (1979). Sprouting of sympathetic nerves in the absence of afferent input. Exp. Neurol, 66, 778–83.CrossRefPubMedGoogle Scholar
  13. Davies, P. (1985). Is it possible to design rational treatment for the symptoms of Alzheimer’s disease? Drug Develop Res., 5, 69–75.Google Scholar
  14. Fonnum, F. (1975). A rapid radiochemical method for the determination of choline acetyltransferase. J. Neurochem., 24, 407–9.CrossRefPubMedGoogle Scholar
  15. Gnahn, H., Hefti, F., Heumann, R., Schwab, M., and Thoenen, H. (1983). NGF-mediated increase of choline acetyltransferase (ChAT) in the neonatal forebrain; evidence for a physiological role of NGF in the brain? Dev. Brain Res., 9, 45–52.Google Scholar
  16. Gorio, A., Marini, P., and Zanoni, R. (1983). Muscle reinnervation. III. Motoneuron sprouting capacity, enhancement by exogenous gangliosides. Neuroscience, 8, 417–29.CrossRefPubMedGoogle Scholar
  17. Grave, G. D. (1977). Thyroid Hormones and Brain Development, Raven Press, New York.Google Scholar
  18. Greene, L. A., and Shooter, E. M. (1980). The nerve growth factor: biochemistry, synthesis and mechanism of action. Ann. Rev. Neurosci., 3, 353–402.CrossRefPubMedGoogle Scholar
  19. Hefti, F. (1983). Alzheimer’s disease caused by a lack of nerve growth factor? Ann. Neurol., 13, 109–10.Google Scholar
  20. Hefti, F. Dravid, A., and Hartikka, J. (1984). Chronic intraventricular injections of nerve growth factor elevate hippocampal choline acetyltransferase activity in adult rats with partial septo-hippocampal lesions. Brain Res., 293, 305–9.CrossRefPubMedGoogle Scholar
  21. Hefti, F., Hartikka, J., and Frick, W. (1985b). Gangliosides alter morphology and growth of astrocytes and increase the activity of choline acetyltransferase in cultures of dissociated septal neurons. J. Neurosci., 5, 2086–94.PubMedGoogle Scholar
  22. Hefti, F., Hartikka, J., Eckenstein, F., Gnahn, H., Heumann, R., and Schwab, M. (1985a). Nerve growth factor (NGF) increases choline acetyltransferase but not survival or fiber growth of cultured septal cholinergic neurons. Neuroscience, 14, 55–68.CrossRefPubMedGoogle Scholar
  23. Honegger, P. and Lenoir, D. (1980a). Nerve growth factor (NGF) stimulation of cholinergic telencephalic neurons in aggregating cell cultures. Dev. Brain. Res., 3, 229–38.CrossRefGoogle Scholar
  24. Honegger, P., and Lenoir, D. (1980b). Triiodothyronine enhancement of neuronal differentiation in aggregating fetal rat brain cells cultured in a chemically defined medium. Brain Res., 199, 425–34.CrossRefPubMedGoogle Scholar
  25. Jorgensen, E. C. (1978). In Li, C. H. (ed.), Hormonal Proteins and Peptides, Vol. 6. Academic Press, New York, pp. 108–204.Google Scholar
  26. Kalaria, R. N., and Prince, A. K. (1985). The effects of neonatal thyroid deficiency on acetylcholine synthesis and glucose oxidation in rat corpus striatum. Dev. Brain Res., 20, 271–9.CrossRefGoogle Scholar
  27. Karpiak, S. E. (1983). Ganglioside treatment improves recovery of alteration behavior after unilateral entorhinal cortex lesion. Exp. Neurol, 81, 330–9.CrossRefPubMedGoogle Scholar
  28. Kiernan, J. A. (1979). Hypotheses concerned with axonal regeneration in the mammalian nervous system. Biol. Rev., 54, 155–97.CrossRefPubMedGoogle Scholar
  29. Kojima, M., Kim, J. S., Uchimurea, H., Hirano, M., Nakahaia, T., and Matsumoto, T. (1981). Effect of thyroidectomy on choline acetyltransferase in rat hypothalamic nuclei. Brain Res., 209, 227–30.CrossRefPubMedGoogle Scholar
  30. Loy, R., and Moore, R. Y. (1977). Anomalous innervation of the hippocampal formation by peripheral sympathetic axons following mechanical injury. Exp. Neurol., 57, 645–50.CrossRefPubMedGoogle Scholar
  31. Mesulam, M. M., Mufson, E. J., Wainer, B. H., and Levey, A. I. (1983). Central cholinergic pathways in the rat: an overview based on an alternative nomenclature (Ch. 1-Ch. 6). Neuroscience, 10, 1185–201.CrossRefPubMedGoogle Scholar
  32. Milner, T. A., Loy, R., and Amaral, D. G. (1983). An anatomical study of the development of the septo-hippocampal projection in the rat. Dev. Brain Res., 8, 343–71.CrossRefGoogle Scholar
  33. Morgan, J. I., and Seifert, W. (1979). Growth factors and gangliosides: a possible new perspective in neuronal growth control. J. Supramolec. Struc., 10, 111–24.CrossRefGoogle Scholar
  34. Nadler, J. V., Mattews, D. A., Cotman, C. W., and Lynch, G. S. (1974). Development of cholinergic innervation in the hippocampal formation of the rat. Dev. Biol., 36, 142–54.CrossRefPubMedGoogle Scholar
  35. Rybak, S., Ginzburg, I., and Yavin, E. (1983). Gangliosides stimulate neurite outgrowth and induce tubulin mRNA accumulation in neural cells. Biochem. Biophys. Res. Commun., 116, 974–80.CrossRefPubMedGoogle Scholar
  36. Schwab, M., Otten, U., Agid, Y., and Thoenen, H. (1979). Nerve growth factor (NGF) in the rat CNS: absence of specific retrograde axonal transport and tyrosine hydroxylase induction in locus coeruleus and substantia nigra. Brain Res., 168, 473–83.CrossRefPubMedGoogle Scholar
  37. Seiler, M., and Schwab, M. E. (1984). Specific retrograde transport of nerve growth factor (NGF) from neocortex to nucleus basalis in the rat. Brain Res., 300, 33–6.CrossRefPubMedGoogle Scholar
  38. Shelton, D. L., and Reichardt, L. F. (1984). Expression of the nerve growth factor gene correlates with the density of sympathetic innervation in effector organs. Proc. Natl Acad. Sci. USA, 81, 7951–5.CrossRefPubMedPubMedCentralGoogle Scholar
  39. Stenevi, U., and Bjorklund, A. (1978). Growth of vascular sympathetic axons into the hippocampus after lesions of the septo-hippocampal pathway; a pitfall in brain lesion studies. Neurosci. Lett., 7, 219–24.CrossRefPubMedGoogle Scholar
  40. Suda, K., Barde, Y. A., and Thoenen, H. (1978). Nerve growth factor in mouse and rat serum: correlation between bioassay and radioimmunoassay determinations. Proc. Natl Acad. Sci. USA, 75, 4042–6.CrossRefPubMedPubMedCentralGoogle Scholar
  41. Thoenen, H., and Barde, Y. A. (1980). Physiology of nerve growth factor. Physiol. Rev., 60, 1284–335.PubMedGoogle Scholar
  42. Toffano, G., Savoini, G., Moroni, G., Lombardi, G., Calza, L., and Agnati, L. F. (1983). GM1 ganglioside treatment reduces dopamine cell body degeneration in the substantia nigra after unilateral hemitranssection in rat. Brain Res., 296, 233–9.CrossRefGoogle Scholar
  43. Toniolo, G., Dunnett, S. B., Hefti, F., and Will, B. (1985). Acetylcholine-rich transplants in the hippocampus: influence of intrinsic growth factors and application of NGF on choline acetyltransferase activity. Brain Res., 345, 141–6.CrossRefPubMedGoogle Scholar
  44. Valcana, T. (1971). Effect of neonatal hypothyroidism on the development of acetylcholinesterase and choline acetyl transferase activity in rat brain. In Ford, D. H. (ed.), Influence of Hormones on the Nervous System. S. Karger, Basel, pp. 174–84.Google Scholar
  45. Wainer, B. H., Levey, A. I., Mufson, E. F., and Mesulam, M. M. (1984). Cholinergic systems in mammalian brain identified with antibodies against choline acetyltransferase. Neurosci. Int., 6, 163–82.Google Scholar

Copyright information

© The Editors and the Contributors 1986

Authors and Affiliations

  • F. Hefti

There are no affiliations available

Personalised recommendations