Detection of Hypoxia-Induced and Iron Depletion-Induced Mitophagy in Mammalian Cells.
Mitochondrial quality and quantity are not only regulated by mitochondrial fusion and fission but also by mitochondria degradation. Mitophagy, an autophagy specific for damaged or unnecessary mitochondria, is believed to be an important pathway for mitochondrial homeostasis. To date, several stimuli are known to induce mitophagy. Some of these stimuli, however, including hypoxia, iron depletion, and nitrogen starvation, induce mild mitophagy, which is difficult to detect through decreased mitochondrial mass. Recently, we have clearly detected mitophagy induced under these conditions using mito-Keima as a reporter. In this chapter, we describe the protocols for induction and detection of hypoxia-induced and iron depletion-induced mitophagy using mito-Keima-expressed cells.
Department of Cellular Physiology, Niigata University Graduate School of Medical and Dental Sciences, 1-757 Asahimachi-dori, Chuo-ku, Niigata, 951-8510, Japan.
Shun-Ichi
SI
Yamashita
Department of Cellular Physiology, Niigata University Graduate School of Medical and Dental Sciences, 1-757 Asahimachi-dori, Chuo-ku, Niigata, 951-8510, Japan. kanki@med.niigata-u.ac.jp.
Tomotake
T
Kanki
D000818:Animals; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D000860:Hypoxia / Q000235:genetics / Q000378:metabolism*; D007501:Iron / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D063306:Mitophagy; D057054:Molecular Imaging; D014161:Transduction, Genetic
D007501:Iron
Autophagy; Deferiprone; Hypoxia; Iron depletion; Keima; Mitochondria; Mitophagy