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Methods to Study Mitochondrial Structure and Function

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Abstract

Cardiac mitochondria are complex highly organized cellular organelles, which play central roles not only in energy homeostasis but also in various biosynthetic, signaling, and cell death pathways. A wide range of methodological approaches have been developed to assess mitochondrial function in the heart. High- and super-resolution fluorescent microscopy has been used to visualize mitochondria and measure number of mitochondrial characteristics. Electron microscopy and electron tomography can visualize not only mitochondrial ultrastructure but also mitochondrial multiprotein complexes at near-atomic resolution. A whole arsenal of modern molecular biological methods has been exploited in analysis of mtDNA and its dynamics. The in vitro spectrophotometric enzyme assays and polarographic measurements of oxygen consumption are commonly utilized to assess mitochondrial function. Noninvasive methods based on magnetic resonance spectroscopy (MRS) have emerged as a powerful tool to study mitochondrial function in vivo in human heart. Electrophoretic techniques, such as 1D-, 2D-PAGE, and BN-PAGE, have proved to be a very sensitive and informative approach to analyze complex content of mitochondrial proteins. Advances in separation and mass spectrometry (MS)-based technologies have led to the identification of a significant number of mitochondrial proteins from various rodent and human tissues, including heart. Animal models are of a great utility for the investigation of mitochondrial functions and their roles in the heart; however, mtDNA gene targeting still presents a significant technical challenge. Great advances in these methodological approaches have fueled progress in our understanding of mitochondrial functional role in heart physiology and pathophysiology.

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Marín-García, J. (2013). Methods to Study Mitochondrial Structure and Function. In: Mitochondria and Their Role in Cardiovascular Disease. Springer, Boston, MA. https://doi.org/10.1007/978-1-4614-4599-9_2

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