Regulation and Quantification of Cellular Mitochondrial Morphology and Content

We summarize the current insights into how mitochondrial content is regulated and discuss experimental strategies to quantify mitochondrial content

Karl Tronstad; Marco Nooteboom; Linn Nilsson; Julie Nikolaisen; Maciek Sokolewicz; Sander Grefte; Ina Pettersen; Sissel Dyrstad; Fredrik Hoel; Peter Willems; Werner Koopman

2014

Scholarcy highlights

  • Mitochondria house enzyme systems for -oxidation of fatty acids, the tricarboxylic acid cycle, ketogenesis and oxidative phosphorylation and as such are key generators of cellular energy in the form of ATP
  • As an example of quantification of mitochondrial content in 2D microscopy images, we have previously developed a high-content strategy for primary human skin fibroblasts
  • With respect to reactive oxygen species signaling, we provided evidence that mitochondrial morphology and function are controlled by endogenous ROS and redox signaling
  • Since mitochondria represent primary sites of both metabolism and cell signaling, they serve as sensors, effectors, and executors in various physiological processes to accommodate contextual influences
  • Studies of mitochondrial content cannot be completely dissociated from the physiological cellular context thatdetermines an observed mitochondrial content change. This means that interpretation of the obtained data needs to consider at least two major questions in order to evaluate the potential effects on mitochondrial physiology: To what extent does the observed results reflect, or relate to, the authentic physiological context in a viable cell or tissue? Is it possible that the observed effect is reciprocally balanced or compensated by other mechanisms in the cell? “mitochondrial number” still might represent a useful descriptor for some applications and tissues, it does not properly reflect the content of mitochondrial biomass since it depends on the specificities of mitochondrial morphology and size, and phenomena such as fusion and fission
  • Computer-assisted 2D and 3D analysis of live-cell microscopy images provides a promising strategy for quantification and statistical analysis of mitochondrial shape and content in an unbiased manner
  • Experimental conditions and the used cell type should be compatible with the applied quantification approach and not be applied without a proper understanding of the biology involved and potential pitfalls

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