Document Detail


Live imaging of cell extrusion from the epidermis of developing zebrafish.
MedLine Citation:
PMID:  21730948     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Homeostatic maintenance of epithelial tissues requires the continual removal of damaged cells without disrupting barrier function. Our studies have found that dying cells send signals to their live neighbors to form and contract a ring of actin and myosin that ejects it out from the epithelial sheet while closing any gaps that might have resulted from its exit, a process termed cell extrusion¹. The optical clarity of developing zebrafish provides an excellent system to visualize extrusion in living epithelia. Here we describe a method to induce and image extrusion in the larval zebrafish epidermis. To visualize extrusion, we inject a red fluorescent protein labeled probe for F-actin into one-cell stage transgenic zebrafish embryos expressing green fluorescent protein in the epidermis and induce apoptosis by addition of G418 to larvae. We then use time-lapse imaging on a spinning disc confocal microscope to observe actin dynamics and epithelial cell behaviors during the process of apoptotic cell extrusion. This approach allows us to investigate the extrusion process in live epithelia and will provide an avenue to study disease states caused by the failure to eliminate apoptotic cells.
Authors:
George T Eisenhoffer; Jody Rosenblatt
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Publication Detail:
Type:  Research Support, N.I.H., Extramural; Video-Audio Media     Date:  2011-06-27
Journal Detail:
Title:  Journal of visualized experiments : JoVE     Volume:  -     ISSN:  1940-087X     ISO Abbreviation:  J Vis Exp     Publication Date:  2011  
Date Detail:
Created Date:  2011-07-06     Completed Date:  2011-07-25     Revised Date:  2011-10-26    
Medline Journal Info:
Nlm Unique ID:  101313252     Medline TA:  J Vis Exp     Country:  United States    
Other Details:
Languages:  eng     Pagination:  -     Citation Subset:  IM    
Affiliation:
Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah.
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MeSH Terms
Descriptor/Qualifier:
Actins / analysis,  metabolism
Animals
Animals, Genetically Modified
Apoptosis / drug effects,  physiology*
Epidermis / cytology*,  drug effects,  metabolism
Gentamicins / pharmacology
Green Fluorescent Proteins / analysis,  biosynthesis,  genetics
Luminescent Proteins / analysis
Microscopy, Confocal / methods
Zebrafish / embryology*
Grant Support
ID/Acronym/Agency:
1 DP2OD002056-01/OD/NIH HHS; 5T32 CA03247-8/CA/NCI NIH HHS; DP2 OD002056-01/OD/NIH HHS
Chemical
Reg. No./Substance:
0/Actins; 0/Gentamicins; 0/Luminescent Proteins; 0/red fluorescent protein; 147336-22-9/Green Fluorescent Proteins; 49863-47-0/antibiotic G 418

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


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