Abstract
Mitochondrial dynamics play an important role in numerous physiological and pathophysiological phenomena in the developing and adult human heart. Alterations in structural aspects of cellular mitochondrial composition as a function of changes in physiology can easily be visualized using fluorescence microscopy. Commonly, mitochondrial location, number, and morphology are reported qualitatively due to the lack of automated and user-friendly computer-based analysis tools. Mitochondrial Quantification using MATLAB (MQM) is a computer-based tool to quantitatively assess these parameters by analyzing fluorescently labeled mitochondria within the cell; in particular, MQM provides numerical information on the number, area, and location of mitochondria within a cell in a time-efficient, automated, and unbiased way. This chapter describes the use of MQM’s capabilities to quantify mitochondrial changes during human pluripotent stem cell (hPSC) differentiation into spontaneously contracting cardiomyocytes (SC-CMs), which follows physiological pathways of human heart development.
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References
Park SJ, Shin JH, Jeong JI et al (2014) Down-regulation of mortalin exacerbates Abeta-mediated mitochondrial fragmentation and dysfunction. J Biol Chem 289:2195–2204
Rana A, Rera M, Walker DW (2013) Parkin overexpression during aging reduces proteotoxicity, alters mitochondrial dynamics, and extends lifespan. Proc Natl Acad Sci U S A 110:8638–8643
Abou-Sleiman PM, Muqit MM, Wood NW (2006) Expanding insights of mitochondrial dysfunction in Parkinson’s disease. Nat Rev Neurosci 7:207–219
Hashimoto M, Rockenstein E, Crews L et al (2003) Role of protein aggregation in mitochondrial dysfunction and neurodegeneration in Alzheimer’s and Parkinson’s diseases. Neuromolecular Med 4:21–36
Castellani R, Hirai K, Aliev G et al (2002) Role of mitochondrial dysfunction in Alzheimer’s disease. J Neurosci Res 70:357–360
Reddy PH, Beal MF (2008) Amyloid beta, mitochondrial dysfunction and synaptic damage: implications for cognitive decline in aging and Alzheimer’s disease. Trends Mol Med 14:45–53
Yoon Y, Galloway CA, Jhun BS et al (2011) Mitochondrial dynamics in diabetes. Antioxid Redox Signal 14:439–457
Lowell BB, Shulman GI (2005) Mitochondrial dysfunction and type 2 diabetes. Science 307:384–387
Desai SP, Bhatia SN, Toner M et al (2013) Mitochondrial localization and the persistent migration of epithelial cancer cells. Biophys J 104:2077–2088
Chen L, Knowlton AA (2011) Mitochondrial dynamics in heart failure. Congest Heart Fail 17:257–261
Ong SB, Hausenloy DJ (2010) Mitochondrial morphology and cardiovascular disease. Cardiovasc Res 88:16–29
Liu S, Bai Y, Huang J et al (2013) Do mitochondria contribute to left ventricular non-compaction cardiomyopathy? New findings from myocardium of patients with left ventricular non-compaction cardiomyopathy. Mol Genet Metab 109:100–106
Dhalla NS, Rangi S, Zieroth S et al (2012) Alterations in sarcoplasmic reticulum and mitochondrial functions in diabetic cardiomyopathy. Exp Clin Cardiol 17:115–120
Lopaschuk GD, Jaswal JS (2010) Energy metabolic phenotype of the cardiomyocyte during development, differentiation, and postnatal maturation. J Cardiovasc Pharmacol 56:130–140
Folmes CD, Dzeja PP, Nelson TJ et al (2012) Metabolic plasticity in stem cell homeostasis and differentiation. Cell Stem Cell 11:596–606
Mitra K, Lippincott-Schwartz J (2010) Analysis of mitochondrial dynamics and functions using imaging approaches. Curr Protoc Cell Biol Chapter 4:Unit 4.25.1–21
Chung S, Dzeja PP, Faustino RS et al (2007) Mitochondrial oxidative metabolism is required for the cardiac differentiation of stem cells. Nat Clin Pract Cardiovasc Med 4(Suppl 1):S60–S67
Hattori F, Chen H, Yamashita H et al (2010) Nongenetic method for purifying stem cell-derived cardiomyocytes. Nat Methods 7:61–66
Rafelski SM (2013) Mitochondrial network morphology: building an integrative, geometrical view. BMC Biol 11:71
Lian X, Zhang J, Azarin SM et al (2013) Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/beta-catenin signaling under fully defined conditions. Nat Protoc 8:162–175
Acknowledgment
This work was supported by AHA 13PRE1470078 (P.K.), NSF-CBET-1150854 (E.A.L.), NSF-EPS-1158862 (D.A.D.), and NSF-EEC-1063107 (K.M.D.). D.A.D. is now located at the Department of Biological Sciences, State University of New York at Oswego, Oswego, NY.
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Kerscher, P., Bussie, B.S., DeSimone, K.M., Dunn, D.A., Lipke, E.A. (2015). Characterization of Mitochondrial Populations During Stem Cell Differentiation. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 1264. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2257-4_37
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DOI: https://doi.org/10.1007/978-1-4939-2257-4_37
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