title: A screening-based platform for the assessment of cellular respiration in Caenorhabditis elegans.
abstract: Mitochondrial dysfunction is at the core of many diseases ranging from inherited metabolic diseases to common conditions that are associated with aging. Although associations between aging and mitochondrial function have been identified using mammalian models, much of the mechanistic insight has emerged from Caenorhabditis elegans. Mitochondrial respiration is recognized as an indicator of mitochondrial health. The Seahorse XF96 respirometer represents the state-of-the-art platform for assessing respiration in cells, and we adapted the technique for applications involving C. elegans. Here we provide a detailed protocol to optimize and measure respiration in C. elegans with the XF96 respirometer, including the interpretation of parameters and results. The protocol takes ∼2 d to complete, excluding the time spent culturing C. elegans, and it includes (i) the preparation of C. elegans samples, (ii) selection and loading of compounds to be injected, (iii) preparation and execution of a run with the XF96 respirometer and (iv) postexperimental data analysis, including normalization. In addition, we compare our XF96 application with other existing techniques, including the eight-well Seahorse XFp. The main benefits of the XF96 include the limited number of worms required and the high throughput capacity due to the 96-well format.
authors: Laboratory of Molecular Neurobiology of Ageing, European Research Institute for the Biology of Ageing, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors: Mandy
authors: M
authors: Koopman
authors: Laboratory of Molecular Neurobiology of Ageing, European Research Institute for the Biology of Ageing, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors: Helen
authors: H
authors: Michels
authors: Laboratory of Cardiac Energetics, National Heart, Lung and Blood Institute, Bethesda, Maryland, USA.
authors: Beverley M
authors: 0000-0002-0896-6024
authors: BM
authors: Dancy
authors: Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, the Netherlands.
authors: Rashmi
authors: R
authors: Kamble
authors: Laboratory of Integrative and Systems Physiology, Ecole Polytechnique Federale de Lausanne, Lausanne, Switzerland.
authors: Laurent
authors: L
authors: Mouchiroud
authors: Laboratory of Integrative and Systems Physiology, Ecole Polytechnique Federale de Lausanne, Lausanne, Switzerland.
authors: Johan
authors: J
authors: Auwerx
authors: Laboratory of Molecular Neurobiology of Ageing, European Research Institute for the Biology of Ageing, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
authors: Ellen A A
authors: EA
authors: Nollen
authors: Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, the Netherlands.
authors: Riekelt H
authors: 0000-0001-9961-0842
authors: RH
authors: Houtkooper
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000295:instrumentation / Q000379:methods; D017173:Caenorhabditis elegans / Q000166:cytology* / Q000378:metabolism; D019069:Cell Respiration; D004867:Equipment Design; D008928:Mitochondria / Q000378:metabolism*; D010101:Oxygen Consumption