Autophagosome and Phagosome pp 125-133

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

Experimental Control and Characterization of Autophagy in Drosophila

  • Gabor Juhasz
  • Thomas P. Neufeld


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.

Key Words

Atg8 autophagy Drosophila GFP TEM LysoTracker Red somatic clones 


  1. 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. 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.CrossRefPubMedGoogle Scholar
  3. 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.CrossRefPubMedGoogle Scholar
  4. 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.CrossRefPubMedGoogle Scholar
  5. 5.
    Martin, D. N., and Baehrecke, E. H. (2004) Caspases function in autophagic programmed cell death in Drosophila. Development 131, 275–284.CrossRefPubMedGoogle Scholar
  6. 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.CrossRefPubMedGoogle Scholar
  7. 7.
    Lee, C. Y., and Baehrecke, E. H. (2001) Steroid regulation of autophagic programmed cell death during development. Development 128, 1443–1455.PubMedGoogle Scholar
  8. 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.CrossRefPubMedGoogle Scholar
  9. 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.CrossRefPubMedGoogle Scholar
  10. 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.CrossRefPubMedGoogle Scholar
  11. 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.CrossRefPubMedGoogle Scholar
  12. 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.CrossRefPubMedGoogle Scholar
  13. 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.CrossRefPubMedGoogle Scholar
  14. 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.CrossRefPubMedGoogle Scholar
  15. 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.CrossRefPubMedGoogle Scholar
  16. 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.CrossRefPubMedGoogle Scholar
  17. 17.
    Theodosiou, N. A., and Xu, T. (1998) Use of FLP/FRT system to study Drosophila development. Methods 14, 355–365.CrossRefPubMedGoogle Scholar
  18. 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.CrossRefPubMedGoogle Scholar
  19. 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.CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Gabor Juhasz
    • 1
  • Thomas P. Neufeld
    • 1
  1. 1.Department of General ZoologyEÖtvÖs Loránd UniversityHungary
  2. 2.Department of Genetics, Cell Biology and DevelopmentUniversity of MinnesotaMN

Personalised recommendations