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In Vivo Analysis of Autophagy in Response to Nutrient Starvation Using Transgenic Mice Expressing a Fluorescent Autophagosome Marker

Mizushima, Noboru, Yamamoto, Akitsugu, Matsui, Makoto, Yoshimori, Tamotsu, Ohsumi, Yoshinori·2004·Molecular Biology of the Cell
DOI 10.1091/mbc.e03-09-0704
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Abstract

Macroautophagy mediates the bulk degradation of cytoplasmic components. It accounts for the degradation of most long-lived proteins: cytoplasmic constituents, including organelles, are sequestered into autophagosomes, which subsequently fuse with lysosomes, where degradation occurs. Although the possible involvement of autophagy in homeostasis, development, cell death, and pathogenesis has been repeatedly pointed out, systematic in vivo analysis has not been performed in mammals, mainly because of a limitation of monitoring methods. To understand where and when autophagy occurs in vivo, we have generated transgenic mice systemically expressing GFP fused to LC3, which is a mammalian homologue of yeast Atg8 (Aut7/Apg8) and serves as a marker protein for autophagosomes. Fluorescence microscopic analyses revealed that autophagy is differently induced by nutrient starvation in most tissues. In some tissues, autophagy even occurs actively without starvation treatments. Our results suggest that the regulation of autophagy is organ dependent and the role of autophagy is not restricted to the starvation response. This transgenic mouse model is a useful tool to study mammalian autophagy.

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  • The presence of yellow puncta suggests autophagic flux is being blocked by inhibition of lysosome fusion with the autophagosome, and can be induced by Bafilomycin A1.

    “The presence of yellow puncta suggests autophagic flux is being blocked by inhibition of lysosome fusion with the autophagosome74–77, and can be induced by the addition of autophagy blocker, Bafilomycin A1 (BAFA1) (Supplementary Fig. 8k).”

    From GRAMD1B is a regulator of lipid homeostasis, autophagic flux and phosphorylated tau (2025) · cites this paper as [77]

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Metadata

Authors
Mizushima, N., Yamamoto, A., Matsui, M., Yoshimori, T., & Ohsumi, Y.
Affiliations
  • Department of Cell Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan
  • Department of Bio-Science, Nagahama Institute of Bio-Science and Technology, Nagahama, Shiga 526-0829, Japan
  • CREST, Japan Science and Technology Agency, Kawaguchi 332-0012, Japan
Venue
Molecular Biology of the Cell
Publisher
American Society for Cell Biology (ASCB)
Published
2004-03
Volume
15
Issue
3
Pages
1101-1111
ISSN
  • 1059-1524
  • 1939-4586
DOI
10.1091/mbc.e03-09-0704
Cited by
2,105
References
5
Language
en

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