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Analysis of gene expression in cultured primary neurons

Fann, Ming-Ji and Patterson, Paul H. (1997) Analysis of gene expression in cultured primary neurons. In: Cellular and Molecular Procedures in Developmental Biology. Current Topics in Developmental Biology. No.36. Academic Press , San Diego, CA, pp. 183-95. ISBN 978-0-12-153136-2. https://resolver.caltech.edu/CaltechAUTHORS:20151229-232049107

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Abstract

This chapter highlights the use of the reverse transcription-polymerase chain reaction (RT-PCR) assay in analyzing the effects of cytokines on neuronal gene expression. Recent progress in molecular biologic techniques has enhanced the utility of primary cultures. Amplification of signals by PCR after RT of RNA to cDNA provides a way to obtain more information than previous methods, and allows the use of small numbers of cultured cells. Furthermore, the RT-PCR may be used to simultaneously determine changes in the expression of multiple neuronal genes. Thus, this approach circumvents some of the limitations of primary culture and greatly enhances the ability to study neuronal gene expression after experimental manipulation. Although the assay introduced in the chapter is only semiquantitative, quantitative results are easily obtained using competitive PCR or Southern analysis after PCR, and making the adjustment of RNA in each sample by calculating ratio of each gene signal to an internal control (such as p-actin). Different ways of preparing neuronal cultures have been introduced and various neuronal stem cells, which are capable of self-replication and generate committed cells, are becoming available.


Item Type:Book Section
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/S0070-2153(08)60502-5DOIArticle
http://www.sciencedirect.com/science/article/pii/S0070215308605025PublisherArticle
Additional Information:© 1998 Academic Press. We thank Doreen McDowell for help with these culture materials and Derek Stemple for advice on designing primers. This project is supported by grants from the NINDS (Javits Neuroscience Investigator Award) to P.H.P., and the National Science Council (Taiwan), R.O.C. to M.-J.F. (NSC84-2331-B010-121).
Funders:
Funding AgencyGrant Number
National Institute of Neurological Disorders and Stroke (NINDS)UNSPECIFIED
National Science Council (Taipei)NSC84-2331-B010-121
Series Name:Current Topics in Developmental Biology
Issue or Number:36
Record Number:CaltechAUTHORS:20151229-232049107
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20151229-232049107
Official Citation:Ming-Ji Fann, Paul H. Patterson, 10 Analysis of Gene Expression in Cultured Primary Neurons, In: Flora de Pablo, Alberto Ferrús and Claudio D. Stern, Editor(s), Current Topics in Developmental Biology, Academic Press, 1997, Volume 36, Pages 183-195, ISSN 0070-2153, ISBN 9780121531362, http://dx.doi.org/10.1016/S0070-2153(08)60502-5. (http://www.sciencedirect.com/science/article/pii/S0070215308605025)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:63262
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:04 Feb 2016 20:44
Last Modified:03 Oct 2019 09:26

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