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Purification and partial characterization of a cholinergic neuronal differentiation factor

Fukada, Keiko (1985) Purification and partial characterization of a cholinergic neuronal differentiation factor. Proceedings of the National Academy of Sciences of the United States of America, 82 (24). pp. 8795-8799. ISSN 0027-8424. PMCID PMC391524. doi:10.1073/pnas.82.24.8795.

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The choice of transmitter made by postmitotic rat sympathetic neurons in cell culture can be controlled by the environment in which they develop. One of the differentiation signals is a protein secreted by heart cells that can induce previously noradrenergic neurons to synthesize acetylcholine and form cholinergic synapses. This change in phenotype occurs without alteration in neuronal survival or growth. The differentiation factor has now been purified at least 100,000-fold, and it is homogeneous by several criteria. (i) The cholinergic activity comigrates with a single 125I-labeled protein band of 45 kDa in one-dimensional NaDodSO4/PAGE. (ii) The biological activity comigrates precisely with a series of five 125I-labeled protein spots of 45 kDa in two-dimensional gel electrophoresis. (iii) Treatment of the 45-kDa band with endo-ß -N-acetylglucosaminidase F reduces the apparent molecular size of both the labeled protein and the biological activity to a band of 22 kDa. The data suggest that the differentiation factor is a slightly basic glycoprotein with at least six glycosylation sites.

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Additional Information:© 1985 by the National Academy of Sciences. Communicated by Masakazu Konishi, August 12, 1985. I wish to thank Paul H. Patterson, in whose laboratory this work was done, for his advice and continual encouragement. I would also like to thank those who provided much of the conditioned medium and nerve growth factor used in this study: Josette Carnahan, Allison Doupe, Joann Imrich, Doreen McDowell, Marie Ryder, Elizabeth Silvestro, Eve Wolinsky, and Vivien Yee. Suggestions that were useful in this work were contributed by Linda Chun, Karina Meiri, Jeremy Brockes, Tadmiri Venkatesh, and David Teplow. The work was supported by fellowships from the Dysautonomia Foundation and the Muscular Dystrophy Association, and by grants to Paul Patterson from the National Institute of Neurological and Communicative Disorders and Stroke, the American Heart Association, and the McKnight and Rita Allen Foundations. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Funding AgencyGrant Number
Dysautonomia FoundationUNSPECIFIED
Muscular Dystrophy AssociationUNSPECIFIED
National Institute of Neurological and Communicative Disorders and Stroke (NINCDS)UNSPECIFIED
American Heart AssociationUNSPECIFIED
McKnight FoundationUNSPECIFIED
Rita Allen FoundationUNSPECIFIED
Subject Keywords:phenotypic choice; sympathetic neuron; development; cardiac cell
Issue or Number:24
PubMed Central ID:PMC391524
Record Number:CaltechAUTHORS:FUKpnas85
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:9186
Deposited By: Tony Diaz
Deposited On:19 Nov 2007
Last Modified:08 Nov 2021 20:57

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