Characterization of Autophagic Responses in Drosophila melanogaster
Abstract
Drosophila is an excellent model system for studying autophagy during animal development due to the availability of genetic reagents and opportunity for in vivo cell biological analysis. The regulation and mechanism of autophagy are highly evolutionarily conserved and the role of autophagy has been characterized during various stages of Drosophila development as well as following starvation. Studies in Drosophila have revealed novel insights into the role of distinct components of the autophagy machinery. This chapter describes protocols for examining autophagy during Drosophila development. A crucial step in the induction of autophagy is the incorporation of Atg8a into the autophagosome. This can be measured as autophagic puncta using live fluorescent imaging, immunostaining, or immunoblot analysis of LC3/Atg8a processing. The level of autophagy can also be examined using other specific components of the autophagy pathway as markers detected by immunofluorescent imaging. Based on the distinct morphology of autophagy, it can also be examined by transmission electron microscopy. In addition, one of the advantages of using Drosophila as a model is the ability to undertake genetic analysis of individual components of the autophagy machinery. Current approaches that can be used to monitor autophagy, including the overall flux and individual steps in Drosophila melanogaster, will be discussed.
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In developing D. melanogaster midgut, caspase inactivation has no consequences, whereas mutations or deletions in essential Atg genes suppress midgut tissue degradation.
“Thus, whereas the genetic inactivation of caspases in the midgut of developing D. melanogaster has no consequences, mutations or deletions in essential autophagy-related (Atg) genes suppress midgut tissue degradation [827–829].”
From Molecular mechanisms of cell death: recommendations of the nomenclature committee on cell death 2018. (2018) · cites this paper as [829]
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- Authors
- Xu, T., Kumar, S., & Denton, D.
- Affiliations
- University of South Australia (AU)
- Corresponding author
- Denton, D.
- Venue
- Methods in Enzymology
- Publisher
- Elsevier
- Published
- 2017
- Volume
- 588
- Pages
- 445-465
- ISSN
- 0076-6879 (ISSN-L)
- OpenAlex
- W2552426574
- Cited by
- 12
- References
- 51
- Citations by year
- 2026: 1 · 2024: 1 · 2022: 1 · 2021: 1 · 2020: 2 · 2019: 2 · 2018: 4
- FWCI
- 10.68
- Type
- book chapter
- Language
- en
- Open access
- closed
- Locations
- Methods in enzymology on CD-ROM/Methods in enzymology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- University of South Australia
- National Institutes of Health
- National Health and Medical Research Council
- National Medical Research Council
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Keywords
- autophagy, Drosophila melanogaster, autophagic flux, Atg8a, autophagosomes, autophagy markers, starvation, autophagy machinery, autophagy monitoring
- MeSH terms
- Autophagy-Related Proteins, Animals, Autophagy, Dissection, Drosophila melanogaster, Microscopy, Fluorescence, Drosophila Proteins, RNA Interference, Microscopy, Electron, Transmission, Gene Knockdown Techniques, Optical Imaging
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