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Mol Cell. 2015 Nov 19;60(4):685-96. doi: 10.1016/j.molcel.2015.10.009. Epub 2015 Nov 5.

Measuring In Vivo Mitophagy.

Author information

1
Center for Molecular Medicine, National Heart Lung and Blood Institute, NIH, Bethesda, MD 20892, USA.
2
Center for Molecular Medicine, National Heart Lung and Blood Institute, NIH, Bethesda, MD 20892, USA; Mitochondria Hub Regulation Center, College of Medicine, Dong-A University, Busan 602-714, Republic of Korea.
3
Light Microscopy Core, National Heart Lung and Blood Institute, NIH, Bethesda, MD 20892, USA.
4
Transgenic Core, National Heart Lung and Blood Institute, NIH, Bethesda, MD 20892, USA.
5
Mitochondria Hub Regulation Center, College of Medicine, Dong-A University, Busan 602-714, Republic of Korea.
6
Center for Molecular Medicine, National Heart Lung and Blood Institute, NIH, Bethesda, MD 20892, USA. Electronic address: finkelt@nih.gov.

Abstract

Alterations in mitophagy have been increasingly linked to aging and age-related diseases. There are, however, no convenient methods to analyze mitophagy in vivo. Here, we describe a transgenic mouse model in which we expressed a mitochondrial-targeted form of the fluorescent reporter Keima (mt-Keima). Keima is a coral-derived protein that exhibits both pH-dependent excitation and resistance to lysosomal proteases. Comparison of a wide range of primary cells and tissues generated from the mt-Keima mouse revealed significant variations in basal mitophagy. In addition, we have employed the mt-Keima mice to analyze how mitophagy is altered by conditions including diet, oxygen availability, Huntingtin transgene expression, the absence of macroautophagy (ATG5 or ATG7 expression), an increase in mitochondrial mutational load, the presence of metastatic tumors, and normal aging. The ability to assess mitophagy under a host of varying environmental and genetic perturbations suggests that the mt-Keima mouse should be a valuable resource.

PMID:
26549682
PMCID:
PMC4656081
DOI:
10.1016/j.molcel.2015.10.009
[Indexed for MEDLINE]
Free PMC Article

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