Mitochondrial Stress Tests Using Seahorse Respirometry on Intact Dictyostelium discoideum Cells.
Mitochondria not only play a critical and central role in providing metabolic energy to the cell but are also integral to the other cellular processes such as modulation of various signaling pathways. These pathways affect many aspects of cell physiology, including cell movement, growth, division, differentiation, and death. Mitochondrial dysfunction which affects mitochondrial bioenergetics and causes oxidative phosphorylation defects can thus lead to altered cellular physiology and manifest in disease. The assessment of the mitochondrial bioenergetics can thus provide valuable insights into the physiological state, and the alterations to the state of the cells. Here, we describe a method to successfully use the Seahorse XF(e)24 Extracellular Flux Analyzer to assess the mitochondrial respirometry of the cellular slime mold Dictyostelium discoideum.
Discipline of Microbiology, La Trobe University, Bundoora, VIC, Australia.
Sui
S
Lay
Discipline of Microbiology, La Trobe University, Bundoora, VIC, Australia.
Oana
O
Sanislav
Department of Physiology Anatomy and Microbiology, Discipline of Microbiology, School of Life Sciences, College of Science Health and Engineering, La Trobe University, Melbourne, VIC, 3086, Australia.
Sarah J
SJ
Annesley
Department of Physiology Anatomy and Microbiology, Discipline of Microbiology, School of Life Sciences, College of Science Health and Engineering, La Trobe University, Melbourne, VIC, 3086, Australia. p.fisher@latrobe.edu.au.
Paul R
PR
Fisher
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D019069:Cell Respiration; D004023:Dictyostelium / Q000378:metabolism*; D004734:Energy Metabolism; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D010085:Oxidative Phosphorylation; D010101:Oxygen Consumption; D023701:Smegmamorpha / Q000378:metabolism*; D013312:Stress, Physiological
D000255:Adenosine Triphosphate
AOX pathway; Dictyostelium discoideum; Matrigel® basement membrane matrix; Mitochondrial inhibitors; Mitochondrial respirometry; OCR; Oxygen consumption rate; Seahorse extracellular flux analyzer; XFe24