Document Detail


Real-time analysis of preprotachykinin promoter activity in single cortical neurons.
MedLine Citation:
PMID:  10899967     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Technological limitations have hindered the study of gene elements regulating transcription within CNS neurons. In the present stuides, rat cortical brain slices endogenously expressing the preprotachykinin (PPT) gene were transfected with gene constructs encompassing green fluorescent protein (GFP) under the control of the PPT promoter. These slices were maintained in organotypic culture so that the fluorescence intensity within individual living cells could be quantified using laser scanning confocal microscopy before and after application of stimulatory agents. Combined treatment with forskolin and elevated potassium significantly increased expression of both endogenous PPT mRNA and the PPT promoter-GFP construct. The ability to follow fluorescence changes within single neurons in real time offers a powerful "within-subject" experimental approach for analysis of neural gene promoters.
Authors:
P D Walker; R Andrade; J P Quinn; M J Bannon
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of neurochemistry     Volume:  75     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  2000 Aug 
Date Detail:
Created Date:  2000-08-17     Completed Date:  2000-08-17     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  882-5     Citation Subset:  IM    
Affiliation:
Department of Anatomy and Cell Biology, Wayne State University School of Medicine, Detroit, Michigan, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Animals, Newborn
Cerebral Cortex / metabolism*
Forskolin / pharmacology
Gene Expression Regulation / drug effects
Green Fluorescent Proteins
Kinetics
Luminescent Proteins / biosynthesis,  genetics
Male
Neurons / drug effects,  metabolism*
Organ Culture Techniques
Potassium / pharmacology
Promoter Regions, Genetic*
Protein Precursors / genetics*
Rats
Recombinant Fusion Proteins / biosynthesis
Tachykinins / genetics*
Time Factors
Transcription, Genetic
Grant Support
ID/Acronym/Agency:
DA06470/DA/NIDA NIH HHS; MH60854/MH/NIMH NIH HHS; NS34935/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Luminescent Proteins; 0/Protein Precursors; 0/Recombinant Fusion Proteins; 0/Tachykinins; 0/preprotachykinin; 147336-22-9/Green Fluorescent Proteins; 66428-89-5/Forskolin; 7440-09-7/Potassium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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