Document Detail


Small-molecule fluorophores to detect cell-state switching in the context of high-throughput screening.
MedLine Citation:
PMID:  18327938     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A small molecule capable of distinguishing the distinct states resulting from cellular differentiation would be of enormous value, for example, in efforts aimed at regenerative medicine. We screened a collection of fluorescent small molecules for the ability to distinguish the differentiated state of a mouse skeletal muscle cell line. High-throughput fluorescence-based screening of C2C12 myoblasts and myotubes resulted in the identification of six compounds with the desired selectivity, which was confirmed by high-content screening in the same cell states. The compound that resulted in the greatest fluorescence intensity difference between the cell states was used as the screening agent in a pilot screen of 84 kinase inhibitors, each present in four doses, for inhibition of myogenesis. Of the kinase inhibitors, 17 resulted in reduction of fluorescence at one or more concentrations; among the "hits" included known inhibitors of myogenesis, confirming that this compound is capable of detecting the differentiated myotube state. We suggest that the strategy of screening for screening agents reported here may be extended more broadly in the future.
Authors:
Bridget K Wagner; Hyman A Carrinski; Young-Hoon Ahn; Yun Kyung Kim; Tamara J Gilbert; Dina A Fomina; Stuart L Schreiber; Young-Tae Chang; Paul A Clemons
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2008-03-08
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  130     ISSN:  1520-5126     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2008 Apr 
Date Detail:
Created Date:  2008-03-26     Completed Date:  2008-04-25     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4208-9     Citation Subset:  IM    
Affiliation:
Chemical Biology Program, Broad Institute of Harvard and MIT, Cambridge, Massachusetts 02142, USA. bwagner@broad.harvard.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Differentiation / physiology*
Cell Line
Drug Evaluation, Preclinical
Enzyme Activation / drug effects
Enzyme Inhibitors / chemistry,  metabolism,  pharmacokinetics
Fluorescent Dyes / chemistry*,  metabolism,  pharmacokinetics
Mice
Microscopy, Fluorescence
Molecular Probes / chemistry*,  metabolism,  pharmacokinetics
Molecular Weight
Muscle Fibers, Skeletal / cytology,  drug effects,  metabolism*
Myoblasts / cytology,  drug effects,  metabolism*
Phosphotransferases / antagonists & inhibitors
Predictive Value of Tests
Time Factors
Grant Support
ID/Acronym/Agency:
DK060837/DK/NIDDK NIH HHS; P20-GM072029/GM/NIGMS NIH HHS; //Howard Hughes Medical Institute
Chemical
Reg. No./Substance:
0/Enzyme Inhibitors; 0/Fluorescent Dyes; 0/Molecular Probes; EC 2.7.-/Phosphotransferases

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


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