Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1977 Dec 1;75(3):712–718. doi: 10.1083/jcb.75.3.712

Role of nerve growth factor in the development of rat sympathetic neurons in vitro. III. Effect on acetylcholine production

PMCID: PMC2111589  PMID: 925077

Abstract

The effect of nerve growth factor (NGF) on the development of cholinergic sympathetic neurons was studied in cultures grown either on monolayers of dissociated rat heart cells or in medium conditioned by them. In the presence of rat heart cells the absolute requirement of neurons for exogenous NGF was partially spared. The ability of heart cells to support neuronal survival was due at least in part to production of a diffusable NGF-like substance into the medium. Although some neurons survived on the heart cell monolayer without added NGF, increased levels of exogenous NGF increased neuronal survival until saturation was achieved at 0.5 microgram/ml 7S NGF. The ability of neurons to produce acetylcholine (ACh) from choline was also dependent on the level of exogenous NGF. In mixed neuron-heart cell cultures, NGF increased both ACh and catecholamine (CA) production per neuron to the same extent; saturation occurred at 1 microgram/ml 7S NGF. As cholinergic neurons developed in culture, they became less dependent on NGF for survival and ACh production, but even in older cultures approximately 40% of the neurons died when NGF was withdrawn. Thus, NGF is as necessary for survival, growth, and differentiation of sympathetic neurons when the neurons express cholinergic functions as when the neurons express adrenergic functions (4, 5).

Full Text

The Full Text of this article is available as a PDF (466.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Angeletti P. U., Levi-Montalcini R., Caramia F. Analysis of the effects of the antiserum to the nerve growth factor in adult mice. Brain Res. 1971 Apr 2;27(2):343–355. doi: 10.1016/0006-8993(71)90259-9. [DOI] [PubMed] [Google Scholar]
  2. Chun L. L., Patterson P. H. Role of nerve growth factor in the development of rat sympathetic neurons in vitro. I. Survival, growth, and differentiation of catecholamine production. J Cell Biol. 1977 Dec;75(3):694–704. doi: 10.1083/jcb.75.3.694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Furshpan E. J., MacLeish P. R., O'Lague P. H., Potter D. D. Chemical transmission between rat sympathetic neurons and cardiac myocytes developing in microcultures: evidence for cholinergic, adrenergic, and dual-function neurons. Proc Natl Acad Sci U S A. 1976 Nov;73(11):4225–4229. doi: 10.1073/pnas.73.11.4225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Johnson M., Ross D., Meyers M., Rees R., Bunge R., Wakshull E., Burton H. Synaptic vesicle cytochemistry changes when cultured sympathetic neurones develop cholinergic interactions. Nature. 1976 Jul 22;262(5566):308–310. doi: 10.1038/262308a0. [DOI] [PubMed] [Google Scholar]
  5. Klingman G. I. The distribution of acetylcholinesterase in sympathetic ganglia of immunosympathectomized rats. J Pharmacol Exp Ther. 1970 May;173(1):205–211. [PubMed] [Google Scholar]
  6. Ko C. P., Burton H., Johnson M. I., Bunge R. P. Synaptic transmission between rat superior cervical ganglion neurons in dissociated cell cultures. Brain Res. 1976 Dec 3;117(3):461–485. doi: 10.1016/0006-8993(76)90753-8. [DOI] [PubMed] [Google Scholar]
  7. Lazarus K. J., Bradshaw R. A., West N. R., Bunge P. Adaptive survival or rat sympathetic neurons cultured without supporting cells or exogenous nerve growth factor. Brain Res. 1976 Aug 20;113(1):159–164. doi: 10.1016/0006-8993(76)90013-5. [DOI] [PubMed] [Google Scholar]
  8. Mains R. E., Patterson P. H. Primary cultures of dissociated sympathetic neurons. I. Establishment of long-term growth in culture and studies of differentiated properties. J Cell Biol. 1973 Nov;59(2 Pt 1):329–345. doi: 10.1083/jcb.59.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nurse C. A., O'Lague P. H. Formation of cholinergic synapses between dissociated sympathetic neurons and skeletal myotubes of the rat in cell culture. Proc Natl Acad Sci U S A. 1975 May;72(5):1955–1959. doi: 10.1073/pnas.72.5.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. O'Lague P. H., Obata K., Claude P., Furshpan E. J., Potter D. D. Evidence for cholinergic synapses between dissociated rat sympathetic neurons in cell culture. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3602–3606. doi: 10.1073/pnas.71.9.3602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Oger J., Arnason B. G., Pantazis N., Lehrich J., Young M. Synthesis of nerve growth factor by L and 3T3 cells in culture. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1554–1558. doi: 10.1073/pnas.71.4.1554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Patterson P. H., Chun L. L. The induction of acetylcholine synthesis in primary cultures of dissociated rat sympathetic neurons. I. Effects of conditioned medium. Dev Biol. 1977 Apr;56(2):263–280. doi: 10.1016/0012-1606(77)90269-x. [DOI] [PubMed] [Google Scholar]
  13. Patterson P. H., Chun L. L. The influence of non-neuronal cells on catecholamine and acetylcholine synthesis and accumulation in cultures of dissociated sympathetic neurons. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3607–3610. doi: 10.1073/pnas.71.9.3607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Patterson P. H., Reichardt L. F., Chun L. L. Biochemical studies on the development of primary sympathetic neurons in cell culture. Cold Spring Harb Symp Quant Biol. 1976;40:389–397. doi: 10.1101/sqb.1976.040.01.037. [DOI] [PubMed] [Google Scholar]
  15. SJOQVIST F. The correlation between the occurrence and localization of acetylcholinesterase-rich cell bodies in the stellate ganglion and the outflow of cholinergic sweat secretory fibres to the fore paw of the cat. Acta Physiol Scand. 1963 Apr;57:339–351. doi: 10.1111/j.1748-1716.1963.tb02597.x. [DOI] [PubMed] [Google Scholar]
  16. Varon S., Raiborn C., Norr S. Association of antibody to nerve growth factor with ganglionic non-neurons (glia) and consequent interference with their neuron-supportive action. Exp Cell Res. 1974 Oct;88(2):247–256. doi: 10.1016/0014-4827(74)90238-9. [DOI] [PubMed] [Google Scholar]
  17. Yamauchi A., Lever J. D., Kemp K. W. Catecholamine loading and depletion in the rat superior cervical ganglion. A formol fluorescence and enzyme histochemical study with numerical assessments. J Anat. 1973 Feb;114(Pt 2):271–282. [PMC free article] [PubMed] [Google Scholar]
  18. Young M., Oger J., Blanchard M. H., Asdourian H., Amos H., Arnason B. G. Secretion of a nerve growth factor by primary chick fibroblast cultures. Science. 1975 Jan 31;187(4174):361–362. doi: 10.1126/science.1167427. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES