Document Detail


Mouse lines with photo-activatable mitochondria to study mitochondrial dynamics.
MedLine Citation:
PMID:  22821887     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Many pathological states involve dysregulation of mitochondrial fusion, fission, or transport. These dynamic events are usually studied in cells lines because of the challenges in tracking mitochondria in tissues. To investigate mitochondrial dynamics in tissues and disease models, we generated two mouse lines with photo-activatable mitochondria (PhAM). In the PhAM(floxed) line, a mitochondrially localized version of the photo-convertible fluorescent protein Dendra2 (mito-Dendra2) is targeted to the ubiquitously expressed Rosa26 locus, along with an upstream loxP-flanked termination signal. Expression of Cre in PhAM( floxed) cells results in bright mito-Dendra2 fluorescence without adverse effects on mitochondrial morphology. When crossed with Cre drivers, the PhAM(floxed) line expresses mito-Dendra2 in specific cell types, allowing mitochondria to be tracked even in tissues that have high cell density. In a second line (PhAM(excised) ), the expression of mito-Dendra2 is ubiquitous, allowing mitochondria to be analyzed in a wide range of live and fixed tissues. By using photo-conversion techniques, we directly measured mitochondrial fusion events in cultured cells as well as tissues such as skeletal muscle. These mouse lines facilitate analysis of mitochondrial dynamics in a wide spectrum of primary cells and tissues, and can be used to examine mitochondria in developmental transitions and disease states.
Authors:
Anh H Pham; J Michael McCaffery; David C Chan
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-08-11
Journal Detail:
Title:  Genesis (New York, N.Y. : 2000)     Volume:  50     ISSN:  1526-968X     ISO Abbreviation:  Genesis     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-11-22     Completed Date:  2013-04-22     Revised Date:  2014-03-19    
Medline Journal Info:
Nlm Unique ID:  100931242     Medline TA:  Genesis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  833-43     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Wiley Periodicals, Inc.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Tracking
Cells, Cultured
Fibroblasts / metabolism
Gene Expression
Green Fluorescent Proteins / biosynthesis,  genetics
Light
Male
Mice
Mice, 129 Strain
Mice, Inbred C57BL
Mice, Transgenic
Microscopy, Fluorescence
Mitochondria, Heart / metabolism,  physiology,  radiation effects
Mitochondrial Dynamics*
Myocytes, Cardiac / metabolism,  radiation effects
Organ Specificity
Primary Cell Culture
Recombinant Proteins / biosynthesis,  genetics
Staining and Labeling
Grant Support
ID/Acronym/Agency:
GM062967/GM/NIGMS NIH HHS; R01 GM062967/GM/NIGMS NIH HHS; //Howard Hughes Medical Institute
Chemical
Reg. No./Substance:
0/Recombinant Proteins; 147336-22-9/Green Fluorescent Proteins
Comments/Corrections

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


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