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Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis

Karbowski, Mariusz and Arnoult, Damien and Chen, Hsuichen and Chan, David C. and Smith, Carolyn L. and Youle, Richard J. (2004) Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis. Journal of Cell Biology, 164 (4). pp. 493-499. ISSN 0021-9525. PMCID PMC2172000. http://resolver.caltech.edu/CaltechAUTHORS:KARjcb04

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Abstract

A dynamic balance of organelle fusion and fission regulates mitochondrial morphology. During apoptosis this balance is altered, leading to an extensive fragmentation of the mitochondria. Here, we describe a novel assay of mitochondrial dynamics based on confocal imaging of cells expressing a mitochondrial matrix–targeted photoactivable green fluorescent protein that enables detection and quantification of organelle fusion in living cells. Using this assay, we visualize and quantitate mitochondrial fusion rates in healthy and apoptotic cells. During apoptosis, mitochondrial fusion is blocked independently of caspase activation. The block in mitochondrial fusion occurs within the same time range as Bax coalescence on the mitochondria and outer mitochondrial membrane permeabilization, and it may be a consequence of Bax/Bak activation during apoptosis.


Item Type:Article
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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2172000/PublisherArticle
http://www.jcb.org/cgi/content/full/jcb.200309082/DC1PublisherSupplemental Material
Additional Information:© 2004 The Rockefeller University Press. Submitted: 11 September 2003. Accepted: 6 January 2004. Published online 9 February 2004. doi:10.1083/jcb.200309082 The authors thank G.H. Patterson and J. Lippincott-Schwartz for PAGFP vectors, C. Blackstone for critical reading of the manuscript, C.A. Winters for neurons, and R. Raju and M. Dalakas for myocytes. Online supplemental material Fig. S1 shows a correlation between the changes in the mitochondrial morphology and the release of endogenous cytochrome c from mitochondria and a conformational change in the endogenous Bax protein. Online supplemental material is available at http://www.jcb.org/cgi/content/full/jcb.200309082/DC1.
Subject Keywords:dynamin; fission; Opal; PAGFP; photoactivation
PubMed Central ID:PMC2172000
Record Number:CaltechAUTHORS:KARjcb04
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:KARjcb04
Alternative URL:http://dx.doi.org/10.1083/jcb.200309082
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:6786
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:21 Dec 2006
Last Modified:07 Jul 2017 18:44

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