Skip to main content

Experimental Control and Characterization of Autophagy in Drosophila

  • Protocol
Autophagosome and Phagosome

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 445))

summary

Insects such as the fruit fly Drosophila melanogaster, which fundamentally reorganize their body plan during metamorphosis, make extensive use of autophagy for their normal development and physiology. In the fruit fly, the hepatic/adipose organ known as the fat body accumulates nutrient stores during the larval feeding stage. Upon entering metamorphosis, as well as in response to starvation, these nutrients are mobilized through a massive induction of autophagy, providing support to other tissues and organs during periods of nutrient deprivation. High levels of autophagy are also observed in larval tissues destined for elimination, such as the salivary glands and larval gut. Drosophila is emerging as an important system for studying the functions and regulation of autophagy in an in vivo setting. In this chapter we describe reagents and methods for monitoring autophagy in Drosophila, focusing on the larval fat body. We also describe methods for experimentally activating and inhibiting autophagy in this system and discuss the potential for genetic analysis in Drosophila to identify novel genes involved in autophagy.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Neufeld, T. P. (2004) Role of autophagy in developmental cell growth and death: insights from Drosophila, in Autophagy (Klionsky, D. J., ed.), Landes Bioscience, Georgetown, pp. 224–232.

    Google Scholar 

  2. Velentzas, A. D., Nezis, I. P., Stravopodis, D. J., Papassideri, I. S., and Margaritis, L. H. (2007) Mechanisms of programmed cell death during oogenesis in Drosophila virilis. Cell Tissue Res. 327, 399–414.

    Article  PubMed  Google Scholar 

  3. Akdemir, F., Farkas, R., Chen, P., et al. (2006) Autophagy occurs upstream or parallel to the apoptosome during histolytic cell death. Development 133,1457–1465.

    Article  CAS  PubMed  Google Scholar 

  4. Juhasz, G., and Sass, M. (2005) Hid can induce, but is not required for autophagy in polyploid larval Drosophila tissues. Eur. J Cell Biol. 84, 491–502.

    Article  CAS  PubMed  Google Scholar 

  5. Martin, D. N., and Baehrecke, E. H. (2004) Caspases function in autophagic programmed cell death in Drosophila. Development 131, 275–284.

    Article  CAS  PubMed  Google Scholar 

  6. Gorski, S. M., Chittaranjan, S., Pleasance, E. D., et al. (2003) A SAGE approach to discovery of genes involved in autophagic cell death. Curr. Biol. 13, 358–363.

    Article  CAS  PubMed  Google Scholar 

  7. Lee, C. Y., and Baehrecke, E. H. (2001) Steroid regulation of autophagic programmed cell death during development. Development 128, 1443–1455.

    CAS  PubMed  Google Scholar 

  8. Lee, C. Y., Clough, E. A., Yellon, P., Teslovich, T. M., Stephan, D. A., and Baehrecke, E. H. (2003) Genome-wide analyses of steroid- and radiation-triggered programmed cell death in Drosophila. Curr. Biol. 13, 350–357.

    Article  CAS  PubMed  Google Scholar 

  9. Scott, R. C., Juhasz, G., and Neufeld, T. P. (2007) Direct induction of autophagy by Atg1 inhibits cell growth and induces apoptotic cell death. Curr. Biol .17, 1–11.

    Article  CAS  PubMed  Google Scholar 

  10. Scott, R. C., Schuldiner, O., and Neufeld, T. P. (2004) Role and regulation of starvation-induced autophagy in the Drosophila fat body. Dev. Cell 7, 167–178.

    Article  CAS  PubMed  Google Scholar 

  11. Lindmo, K., Simonsen, A., Brech, A., Finley, K., Rusten, T. E., and Stenmark, H. (2006) A dual function for Deep orange in programmed autophagy in the Drosophila melanogaster fat body. Exp. Cell Res. 312, 2018–2027.

    Article  CAS  PubMed  Google Scholar 

  12. Rusten, T. E., Lindmo, K., Juhasz, G., et al. (2004) Programmed autophagy in the Drosophila fat body is induced by ecdysone through regulation of the PI3 K pathway. Dev. Cell 7, 179–192.

    Article  CAS  PubMed  Google Scholar 

  13. Ravikumar, B., Vacher, C., Berger, Z., et al. (2004) Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease. Nat. Genet. 36, 585–595.

    Article  CAS  PubMed  Google Scholar 

  14. Scott, P. H., Brunn, G. J., Kohn, A. D., Roth, R. A., and Lawrence, J. C., Jr. (1998) Evidence of insulin-stimulated phosphorylation and activation of the mammalian target of rapamycin mediated by a protein kinase B signaling pathway. Proc. Natl. Acad. Sci. USA 95, 7772–7777.

    Article  CAS  PubMed  Google Scholar 

  15. Sass, M., and Kovacs, J. (1977) The effect of ecdysone on the fat body cells of the penultimate larvae of Mamestra brassicae. Cell Tissue Res. 180, 403–409.

    Article  CAS  PubMed  Google Scholar 

  16. McGuire, S. E., Roman, G., and Davis, R. L. (2004) Gene expression systems in Drosophila: a synthesis of time and space. Trends Genet. 20, 384–391.

    Article  CAS  PubMed  Google Scholar 

  17. Theodosiou, N. A., and Xu, T. (1998) Use of FLP/FRT system to study Drosophila development. Methods 14, 355–365.

    Article  CAS  PubMed  Google Scholar 

  18. Hennig, K. M., Colombani, J., and Neufeld, T. P. (2006) TOR coordinates bulk and targeted endocytosis in the Drosophila melanogaster fat body to regulate cell growth. J. Cell. Biol. 173, 963–974.

    Article  CAS  PubMed  Google Scholar 

  19. Britton, J. S., Lockwood, W. K., Li, L., Cohen, S. M., and Edgar, B. A. (2002) Drosophila‘s insulin/PI3-kinase pathway coordinates cellular metabolism with nutritional conditions. Dev. Cell 2, 239–429.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Juhasz, G., Neufeld, T.P. (2008). Experimental Control and Characterization of Autophagy in Drosophila. In: Deretic, V. (eds) Autophagosome and Phagosome. Methods in Molecular Biology™, vol 445. Humana Press. https://doi.org/10.1007/978-1-59745-157-4_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-157-4_8

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-853-9

  • Online ISBN: 978-1-59745-157-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics