Expression of Noradrenergic and Cholinergic Traits by Sympathetic Neurons Cultured without Serum
Abstract
The ability to vary systematically the neuronal environment is one advantage afforded by the use of cell culture. Replacement of serum, a variable and undefined medium supplement, with known ingredients allows even greater control of culture conditions. We have studied biochemical and morphological properties related to neurotransmitter metabolism of rat sympathetic neurons cultured in a modified defined medium. Neuronal survival, ultrastructure, and expression of noradrenergic properties appear similar in serum-free and serum-supplemented cultures: small granular vesicles characteristic of norepinephrine storage were observed in both types of culture, and tyrosine hydroxylase activity, conversion of dopamine to norepinephrine, catecholamine production, and storage capacity are equivalent in serum-free and serum-containing cultures. Several of these properties were not exhibited at high levels in previous formulations of this defined medium. Acetylcholine production, however, was about 10-fold lower in serum-free compared to serum-supplemented cultures, consistent with the findings of lacovitti et al. (lacovitti, L., M. I. Johnson, T. H. Joh, and R. P. Bunge (1982) Neuroscience 7:2225–2239). Acetylcholine production can be induced under serum-free conditions by a previously characterized cholinergic inducing factor from heart cell conditioned medium. This responsiveness to serum-free heart cell conditioned medium indicates that serum-free cultures retain plasticity with respect to transmitter status, despite expression of noradrenergic characteristics, unlike cultured neurons of which the noradrenergic transmitter status is maintained by chronic depolarization. Thus, sympathetic neurons survive, express numerous differentiated properties, and display a novel transmitter status under serum-free conditions.
Additional Information
© 1985 by Society for Neuroscience. For the first six months after publication SfN's license will be exclusive. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). Received July 2, 1984; Revised October 5, 1984; Accepted October 8, 1984. We wish to thank Doreen McDowell for assistance with the cell culturing and John Fredieu for assistance with the electron microscopy. We also thank Dr. T. M. Jessel! for helpful discussion of the manuscript. E. J. W. was a predoctoral trainee of the National Institute of General Medical Sciences. S. C. L. is an Established Investigator of the American Heart Association, supported, in part, by the Massachusetts Affiliate. P. H. P. was a Rita Allen Foundation Scholar and a McKnight Foundation Neuroscience Development Awardee. Support was also provided by grants from the National Institute of Neurological and Communicative Disorders and Stroke to P. H. P. and S. C. L.Attached Files
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Additional details
- PMCID
- PMC6565262
- Eprint ID
- 67148
- Resolver ID
- CaltechAUTHORS:20160517-081539389
- NIH Predoctoral Fellowship
- American Heart Association, Massachusetts Affiliate
- Rita Allen Foundation Scholar
- McKnight Foundation
- National Institute of Neurological and Communicative Disorders and Stroke (NINCDS)
- Created
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2016-05-17Created from EPrint's datestamp field
- Updated
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2021-11-11Created from EPrint's last_modified field