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JoVE Journal Neuroscience
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Visualization of Mitochondrial DNA Replication in Individual Cells by EdU Signal Amplification

Article DOI: 10.3791/2147-v • 09:37 min • November 15th, 2010
November 15th, 2010 •
1, 2, 3
1Michigan Research Community, Undergraduate Research Opportunity Program, University of Michigan, 2Department of Neurology, University of Michigan, 3Department of Internal Medicine, Division of Metabolism, Endocrinology, and Diabetes, University of Michigan

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Summary
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We developed a sensitive technique to label newly synthesized mitochondrial DNA (mtDNA) in individual cells in order to study mtDNA biogenesis. The technique combines the incorporation of EdU together with a tyramide signal amplification (TSA) protocol to visualize mtDNA replication within subcellular compartments of neurons.

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Visualization Mitochondrial DNA Replication Cells EdU Signal Amplification Mitochondria Cellular Energy Mitochondrial Biogenesis MtDNA Replication Rate Sensitive Technique Labeling Newly Synthesized MtDNA Individual Cells MtDNA Biogenesis Study 5-ethynyl-2'-deoxyuridine (EdU) Tyramide Signal Amplification (TSA) Visualization Of MtDNA Replication Subcellular Compartments Neurons Thymidine Analogs 5-bromo-2-deoxyuridine (BrdU) Click Reaction Acid Treatments Enzyme Digests EdU Incorporation Comparison Cellular Markers Mechanisms Of Mitochondrial Biogenesis Regulation Pathogenesis Associated With Drug Toxicity Aging Cancer Neurodegenerative Diseases
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