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Small Intensely Fluorescent Cells in Culture: Role of Glucocorticoids and Growth Factors in Their Development and lnterconversions with other Neural Crest Derivatives

Doupe, Allison J. and Patterson, Paul H. and Landis, Story C. (1985) Small Intensely Fluorescent Cells in Culture: Role of Glucocorticoids and Growth Factors in Their Development and lnterconversions with other Neural Crest Derivatives. Journal of Neuroscience, 5 (8). pp. 2143-2160. ISSN 0270-6474. PMCID PMC6565283. doi:10.1523/jneurosci.05-08-02143.1985.

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The neural crest gives rise to a number of adrenergic derivatives, including sympathetic neurons and adrenal chromaffin cells, which contain catecholamines (CAs) but differ in other morphological and functional characteristics. Small intensely fluorescent (SIF) cells, which exist primarily as a minority cell population in autonomic ganglia, are a third cell type in the sympathoadrenal branch of the neural crest lineage. In some respects these cells appear intermediate in phenotype between sympathetic neurons and adrenal chromaffin cells. We established pure dissociated cell cultures of SIF cells from rat superior cervical ganglia (SCG) and used these to study the role of environmental factors in SIF cell development and the relationship of these cells to the other cell types of the sympathoadrenal lineage. When cells from neonatal rat SCG were grown for 3 weeks in the presence of glucocorticoid and in the absence of nerve growth factor (NGF), pure cultures of SIF cells developed. The properties of the cells included (i) small cell size and the occasional presence of short neurites, (ii) intense CA histofluorescence and immunoreactivity for CA synthetic enzymes, (iii) synthesis and storage of CA from radioactive precursors, and (iv) characteristic ultrastructure. The concentration of the glucocorticoid and the presence or absence of non-neuronal cell factors influenced which types of SIF cells developed. In micromolar glucocorticoid most of the cells resembled adrenal chromaffin or type II SIF cells: they displayed immunohistochemically detectable phenylethanolamine-N-methyltransferase (PNMT), synthesized and stored epinephrine, and contained large granular vesicles (100 to 300 nm). When SCG cells were grown in 10(-8) M hormone, many fewer SIF cells developed and a higher percentage of these lacked PNMT immunoreactivity, had neurites, and contained vesicles of smaller mean diameter (70 to 130 nm), similar to those of type I SIF cells in vivo. In the presence of conditioned medium (medium conditioned by non- neuronal cells) as well as glucocorticoid, virtually all of the cells morphologically resembled type I SIF cells. In the absence of glucocorticoid, no SIF cells were ever observed after 3 weeks in culture. By following the development of CA histofluorescence and SIF cell ultrastructure in the cultures over time, we demonstrated that SIF cells were not present in large numbers in these cultures immediately after plating, but were induced from an apparently undifferentiated precursor by the hormonal environment, whereas most of the principal neurons died.

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Additional Information:© 1985 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 ( Received September 4, 1984: Revised December 7, 1984; Accepted December 7, 1984. A. J. D. was a member of and was supported in part by the Harvard University Society of Fellows. and in part by the Insurance Medical Scientist Scholarship Fund and the Prudential Life lnsurtance Co. 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. We wish to thank Doreen MacDowell and John Fredieu for expert technical assistance, and Wendy Brooks, Joe Gagliardi, and Shirley Wilson for manuscript preparation. We also wish to thank Tom Fox for the generous use of his HPLC equipment, and Jane Dodd, Jim Huettner, Tom Jessell, Bill Matthew, and Eve Wolinsky for helpful discussions of the manuscript.
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Harvard UniversityUNSPECIFIED
Prudential Life lnsurtance Co.UNSPECIFIED
American Heart Association, Massachusetts AffiliateUNSPECIFIED
Rita Allen FoundationUNSPECIFIED
McKnight FoundationUNSPECIFIED
Insurance Medical Scientist Scholarship FundUNSPECIFIED
Issue or Number:8
PubMed Central ID:PMC6565283
Record Number:CaltechAUTHORS:20160517-102510768
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Official Citation:Small intensely fluorescent cells in culture: role of glucocorticoids and growth factors in their development and interconversions with other neural crest derivatives AJ Doupe, PH Patterson, and SC Landis The Journal of Neuroscience, 1 August 1985, 5(8):2143-2160
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:67155
Deposited By: Ruth Sustaita
Deposited On:17 May 2016 17:35
Last Modified:11 Nov 2021 00:28

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