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Published October 15, 1977 | public
Journal Article

The induction of acetylcholine synthesis in primary cultures of dissociated rat sympathetic neurons

Abstract

Dissociated sympathetic neurons from the neonatal rat, grown in cell culture in the virtual absence of other cell types, can develop many of the properties expected of differentiated adrenergic neurons including the ability to synthesize and accumulate catecholamines (CA)^2. However, in the presence of high concentrations of appropriately conditioned medium (CM), the cultures develop the ability to synthesize and accumulate acetylcholine (ACh); correspondingly, their ability to synthesize CA decreases. In this paper several developmental aspects of the CM effect are described. The time course of development of cultures grown with or without CM was followed using synthesis and accumulation of [^3H]CA from [^3H]tyrosine and production of [^3H]ACh from [^3H]choline as assays for adrenergic and cholinergic differentiation. The ability to produce CA or ACh developed along parallel time courses in the two sets of cultures, rising primarily during the second week in vitro and reaching a plateau during the fourth week. When CM was used as a cholinergic developmental signal, the sympathetic neurons showed a decreasing response to addition of CM as they matured adrenergically; addition of CM during the third or fourth 10 days in vitro was not as effective in inducing ACh production as addition during the first or second 10 days. Similarly, removal of CM at various times from cultures previously grown in CM showed that the cholinergic induction caused by CM was not easily reversible in older cultures. Thus, as with the adrenergic decision, the cholinergic decision becomes less reversible as the phenotype becomes fully expressed.

Additional Information

© 1977 by Academic Press, Inc. Received 1 June 1977, Accepted 29 June 1977, Available online 8 December 2004. It is a pleasure to acknowledge the superb assistance of Doreen McDowell and Karen Fischer with the culturing. We also thank Eleanor Livingston and Joe Gagliardi for excellent help with the manuscript and Ed Kravitz for the use of his equipment. This work was supported by a grant in aid (93-877) from the American and Massachusetts Heart Associations and a USPHS grant (1 TOI NS 11027) from the National Institute of Neurological and Communicable Diseases and Stroke. PHP is a Research Career Development Awardee of the NINCDS (l K04 NS 70806), and LL YC was a USPHS Trainee (TOl NS 05731).

Additional details

Created:
August 19, 2023
Modified:
October 17, 2023