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Analyzing Mitochondrial Dynamics in Mouse Organotypic Slice Cultures

Pham, Anh H. and Chan, David C. (2014) Analyzing Mitochondrial Dynamics in Mouse Organotypic Slice Cultures. In: Mitochondrial Function. Methods in Enzymology. No.547. Academic Press , Amsterdam, pp. 111-129. ISBN 9780128016152. http://resolver.caltech.edu/CaltechAUTHORS:20141201-094500370

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Abstract

Mitochondria are mobile organelles that dynamically remodel their membranes and actively migrate along cytoskeletal tracks. There is overwhelming evidence that regulators of mitochondrial dynamics are critical for the survival and function of neural tissues. In multiple animal models, ablation of genes regulating mitochondrial shape result in stunted neural development and neurodegeneration. Organotypic cultures serve as ideal in vitro tissue models to further dissect the mechanisms of mitochondrial function in neuronal survival. Slice cultures preserve the three-dimensional cytoarchitecture of neural networks and can survive for prolonged periods in culture. In addition, these cultures allow long-term assessment of genetic or pharmacologic perturbations on neuronal function. Organotypic preparations from murine and rat models have been developed for many regions of the brain. In this chapter, we describe our methods for preparing basal ganglia and cerebellar slice cultures suitable for studying mitochondrial function in Parkinson's disease and cerebellar ataxia, respectively. With such slices, we describe a robust method for live imaging of mitochondrial dynamics. To quantitatively analyze mitochondrial motility, we show how to generate kymographs using the open source image analysis program ImageJ. These techniques provide a powerful platform for assessing mitochondrial activity in neural networks.


Item Type:Book Section
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/B978-0-12-801415-8.00007-2DOIArticle
http://www.sciencedirect.com/science/article/pii/B9780128014158000072PublisherArticle
Additional Information:© 2014 Elsevier Inc.
Subject Keywords:Organotypic cultures; Mitochondrial dynamics; Basal ganglia parasagittal cultures; Cerebellar slice cultures; Kymographs; Live imaging of mitochondria
Record Number:CaltechAUTHORS:20141201-094500370
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20141201-094500370
Official Citation:Anh H. Pham, David C. Chan, Chapter Seven - Analyzing Mitochondrial Dynamics in Mouse Organotypic Slice Cultures, In: Anne N. Murphy and David C. Chan, Editor(s), Methods in Enzymology, Academic Press, 2014, Volume 547, Pages 111-129, ISSN 0076-6879, ISBN 9780128014158, http://dx.doi.org/10.1016/B978-0-12-801415-8.00007-2. (http://www.sciencedirect.com/science/article/pii/B9780128014158000072)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:52207
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:01 Dec 2014 17:55
Last Modified:01 Dec 2014 17:55

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