Document Detail


Purification of cardiac cells from Drosophila embryos.
MedLine Citation:
PMID:  22119843     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recent genome-level innovations have enabled the identification of the entire cadre of genes that are expressed in specific tissues at particular developmental times. However, to be informative as to how individual cell types develop, this process relies upon the successful and efficient purification of cells for a particular tissue. Here, we describe a method to isolate cardiac cells from Drosophila embryos. We generated transgenic embryos in which a cardiac-specific enhancer of the Sulphonylurea receptor (Sur) gene drove expression of the green fluorescent protein (GFP) gene. Homogenized embryos were subjected to fluorescence activated cell sorting (FACS), resulting in approximately 50,000 cardiac cells purified. The prevalence of cardiac cells in the purified population was high, based upon a significant enrichment for cardiac-specific marker genes, including Sur and Toll. This enrichment also enabled the identification of cardiac-specific alternatively spliced isoforms of the Zasp66 gene. In the future, this approach can be used to describe the cardiac transcriptome of Drosophila at distinct stages of embryonic development.
Authors:
Anton L Bryantsev; Richard M Cripps
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2011-11-20
Journal Detail:
Title:  Methods (San Diego, Calif.)     Volume:  56     ISSN:  1095-9130     ISO Abbreviation:  Methods     Publication Date:  2012 Jan 
Date Detail:
Created Date:  2012-03-12     Completed Date:  2012-07-02     Revised Date:  2014-09-18    
Medline Journal Info:
Nlm Unique ID:  9426302     Medline TA:  Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  44-9     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Elsevier Inc. All rights reserved.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Markers / metabolism
Drosophila / cytology*,  embryology*
Drosophila Proteins / genetics
Embryo, Nonmammalian
Flow Cytometry
Myocardium / cytology
Polymerase Chain Reaction
Protein Isoforms / genetics
Toll-Like Receptors / genetics
Grant Support
ID/Acronym/Agency:
HL080545/HL/NHLBI NIH HHS; R01 HL080545/HL/NHLBI NIH HHS; R01 HL080545-05/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Biological Markers; 0/Drosophila Proteins; 0/Protein Isoforms; 0/Tl protein, Drosophila; 0/Toll-Like Receptors
Comments/Corrections

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