Abstract
Peroxisomes and mitochondria are essential subcellular organelles in mammals. Interestingly, recent studies have elucidated that these highly dynamic and plastic organelles exhibit a much closer interrelationship than previously assumed. Peroxisomes and mitochondria are metabolically linked organelles, which are cooperating and cross-talking. They share key components of their division machinery and cooperate in antiviral signaling and defense. As peroxisomal alterations in metabolism, biogenesis, dynamics, and proliferation have the potential to influence mitochondrial morphology and functions (and vice versa), there is currently great interest in the detection of both organelles under different experimental conditions. Here, we present protocols used successfully in our laboratory for the dual detection of peroxisomes and mitochondria in cultured mammalian cells. We address double immunofluorescence and fluorescence-based techniques as well as reagents to investigate organelle dynamics, morphological alterations, and organelle-specific targeting of proteins. In addition, we describe the application of diaminobenzidine cytochemistry on cultured cells to specifically label peroxisomes in ultrastructural studies.
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References
Camoes F et al (2009) Organelle dynamics and dysfunction: a closer link between peroxisomes and mitochondria. J Inherit Metab Dis 32:163–180
Schrader M (2006) Shared components of mitochondrial and peroxisomal division. Biochim Biophys Acta 1763:531–541
Wanders RJA, Waterham HR (2006) Biochemistry of mammalian peroxisomes revisited. Annu Rev Biochem 75:295–332
Dixit E et al (2010) Peroxisomes are signaling platforms for antiviral innate immunity. Cell 141:668–681
Braschi E et al (2010) Vps35 mediates vesicle transport between the mitochondria and peroxisomes. Curr Biol 20:1310–1315
Neuspiel M et al (2008) Cargo-selected transport from the mitochondria to peroxisomes is mediated by vesicular carriers. Curr Biol 18:102–108
Delille HK, Alves R, Schrader M (2009) Biogenesis of peroxisomes and mitochondria: linked by division. Histochem Cell Biol 131:441–446
Nagotu S, Veenhuis M, van der Klei IJ (2010) Divide et impera: the dictum of peroxisomes. Traffic 11:175–184
Waterham HR et al (2007) A lethal defect of mitochondrial and peroxisomal fission. N Engl J Med 356:1736–1741
Wiemer EA et al (1997) Visualization of the peroxisomal compartment in living mammalian cells: dynamic behavior and association with microtubules. J Cell Biol 136:71–80
Koch A et al (2004) Peroxisome elongation and constriction but not fission can occur independently of dynamin-like protein 1. J Cell Sci 117:3995–4006
Fahimi HD (1968) Cytochemical localization of peroxidase activity in rat hepatic microbodies (peroxisomes). J Histochem Cytochem 16:547–550
Fahimi HD (1969) Cytochemical localization of peroxidatic activity of catalase in rat hepatic microbodies (peroxisomes). J Cell Biol 43:275–288
Hirai K (1968) Specific affinity of oxydized amine dye (radical intermediates) for heme enzymes: study in microscopy and spectrophotometry. Acta Histochem Cytochem 1:43
Novikoff AB, Goldfischer S (1968) Visualization of microbodies for light and electron microscopy. J Histochem Cytochem 16:507
Novikoff AB, Goldfischer S (1969) Visualization of peroxisomes (microbodies) and mitochondria with diaminobenzidine. J Histochem Cytochem 17:675–680
Seligman AM et al (1968) Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol 38:1–14
Angermüller S (1989) Peroxisomal oxidases: cytochemical localization and biological relevance. Prog Histochem Cytochem 20:1–65
Fujimoto T, Inomata K, Ogawa K (1982) A cerium method for the ultracytochemical localization of monoamine oxidase activity. Histochem J 14:87–98
Angermuller S, Loffler M (1995) Localization of dihydroorotate oxidase in myocardium and kidney cortex of the rat. An electron microscopic study using the cerium technique. Histochem Cell Biol 103:287–292
Schrader M, Krieglstein K, Fahimi HD (1998) Tubular peroxisomes in HepG2 cells: selective induction by growth factors and arachidonic acid. Eur J Cell Biol 75:87–96
Peranen J, Rikkonen M, Kaariainen L (1993) A method for exposing hidden antigenic sites in paraformaldehyde-fixed cultured cells, applied to initially unreactive antibodies. J Histochem Cytochem 41:447–454
Schrader M et al (1998) Expression of PEX11beta mediates peroxisome proliferation in the absence of extracellular stimuli. J Biol Chem 273:29607–29614
Gehrmann W, Elsner M (2011) A specific fluorescence probe for hydrogen peroxide detection in peroxisomes. Free Radic Res 45:501–506
Ivashchenko O et al (2011) Intraperoxisomal redox balance in mammalian cells: oxidative stress and interorganellar cross-talk. Mol Biol Cell 22:1440–1451
Hanson GT et al (2004) Investigating mitochondrial redox potential with redox-sensitive green fluorescent protein indicators. J Biol Chem 279:13044–13053
Huybrechts SJ et al (2009) Peroxisome dynamics in cultured mammalian cells. Traffic 10:1722–1733
Acknowledgements
We would like to thank H. D. Fahimi (University of Heidelberg, Germany) for stimulating discussions and B. Agricola (University of Marburg, Germany) for excellent technical support. This work was supported by the Portuguese Foundation for Science and Technology (FCT) and FEDER (PTDC/BIA-BCM/099613/2008, PTDC/SAU-OSM/103647/2008, SFRH/BD/37647/2007 (to N.A.B.), SFRH/BPD/74428/2010 (to M.I.), SFRH/BPD/37725/2007 (to M.G. L)), and the University of Aveiro.
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Bonekamp, N.A., Islinger, M., Lázaro, M.G., Schrader, M. (2012). Cytochemical Detection of Peroxisomes and Mitochondria. In: Taatjes, D., Roth, J. (eds) Cell Imaging Techniques. Methods in Molecular Biology, vol 931. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-056-4_24
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DOI: https://doi.org/10.1007/978-1-62703-056-4_24
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