title: Cytochemical detection of peroxisomes and mitochondria.
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.
authors.affiliation: Center for Cell Biology and Department of Biology, University of Aveiro, Aveiro, Portugal.
authors.forename: Nina A
authors.initials: NA
authors.lastname: Bonekamp
authors.forename: Markus
authors.initials: M
authors.lastname: Islinger
authors.forename: Maria Gómez
authors.initials: MG
authors.lastname: Lázaro
authors.forename: Michael
authors.initials: M
authors.lastname: Schrader
mesh_terms: D015100:3,3'-Diaminobenzidine / Q000737:chemistry; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure*; D020675:Peroxisomes / Q000378:metabolism / Q000648:ultrastructure*; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013194:Staining and Labeling
chemical_list: D007202:Indicators and Reagents; D008565:Membrane Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D015100:3,3'-Diaminobenzidine