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The Induction of Acetylcholine Synthesis in Primary Cultures of Dissociated Rat Sympathetic Neurons I. Effects of Conditioned Medium

Patterson, Paul H. and Chun, Linda L. Y. (1977) The Induction of Acetylcholine Synthesis in Primary Cultures of Dissociated Rat Sympathetic Neurons I. Effects of Conditioned Medium. Developmental Biology, 56 (2). pp. 263-280. ISSN 0012-1606. http://resolver.caltech.edu/CaltechAUTHORS:20160728-115103511

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Abstract

Neurotransmitter metabolism in primary cultures of dissociated sympathetic neurons from the newborn rat has been examined. Previous studies have shown that neurons grown in the virtual absence of nonneuronal cells developed the ability to synthesize and accumulate radioactive norepinephrine (NE) from [^3H]tyrosine, but synthesized little [^3H]acetylcholine (ACh) from labeled choline. However, in the presence of certain types of nonneuronal cells, or in medium conditioned (CM) by them, the neurons produced considerable ACh from choline. Here we show that ACh production from choline depends on the concentration of CM in the growth medium. At 62% CM, the highest level of CM studied, ACh production is >40-fold higher than the control value. Furthermore, homogenates of mixed neuronal/nonneuronal cultures or CM-treated neuronal cultures contain 100- to 1000-fold more choline acetyltransferase activity than neuron-alone sister cultures. On the other hand, the ability to synthesize and accumulate NE is inversely related to the percentage CM in the growth medium; at high concentrations of CM, catecholamine production is depressed about 25-fold from the control. The effects of CM are rather specific, since CM does not affect the number of neurons surviving or their overall growth. These results suggest that CM acts by altering the differentiated fate of individual sympathetic neurons rather than by causing the selective survival or growth of a second population of neurons. CM from primary cultures of several tissues taken from the newborn rat induces neuronal ACh synthesis, as does CM from cultures of several, but not all types of rat cell lines. In contrast, CM from several non-rat cell lines as well as from primary cultures of mouse and chick heart is less effective in causing neuronal ACh production, suggesting some degree of species specificity in this phenomenon.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/0012-1606(77)90269-XDOIArticle
http://www.sciencedirect.com/science/article/pii/001216067790269XPublisherArticle
Additional Information:© 1977 Elsevier Inc. Received 27 September 1976, Accepted 27 November 1976. It is a pleasure to acknowledge the conscientious assistance of Doreen McDowell and Karen Fischer with the culturing. We also thank Eleanor P. Livingston, Shirley E. Wilson, Joe Gagliardi, and Zach Hall for excellent help with the manuscript, Ed Kravitz for the use of his equipment, Darwin Berg for help with the choline acetyltransferase assays, and Dick Mains for assistance with the initial experiments. This research was supported by a grant-in-aid (93-877) from the American and Massachusetts Heart Associations and a USPHS Grant (1 R0l NS 11027) from the National Institute of Neurological and Communicable Diseases and Stroke. P.H.P. is a Research Career Development Awardee of the NINCDS (1 K04 NS 70806) and L.L.Y.C. is a PHS Trainee (T0l NS 05731).
Funders:
Funding AgencyGrant Number
American Heart Association93-877
U.S. Public Health Service (USPHS)1-R01-NS 11027
NIH1 K04 NS 70806
NIH Predoctoral FellowshipT01 NS 05731
Massachusetts Heart AssociationUNSPECIFIED
Record Number:CaltechAUTHORS:20160728-115103511
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20160728-115103511
Official Citation:Paul H. Patterson, Linda L.Y. Chun, The induction of acetylcholine synthesis in primary cultures of dissociated rat sympathetic neurons: I. Effects of conditioned medium, Developmental Biology, Volume 56, Issue 2, April 1977, Pages 263-280, ISSN 0012-1606, http://dx.doi.org/10.1016/0012-1606(77)90269-X. (http://www.sciencedirect.com/science/article/pii/001216067790269X)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:69283
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:28 Jul 2016 19:51
Last Modified:28 Jul 2016 19:51

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