Skip to main content

Assays to Monitor Aggrephagy in Drosophila Brain

  • Protocol
  • First Online:
Autophagy in Differentiation and Tissue Maintenance

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1854))

Abstract

Accumulation of ubiquitinated protein aggregates is a hallmark of most aging-related neurodegenerative disorders. Autophagy has been found to be involved in the selective clearance of these protein aggregates, and this process is called aggrephagy. Here we provide two protocols for the investigation of protein aggregation and their removal by autophagy using western blotting and immunofluorescence techniques in Drosophila brain. Investigating the role of aggrephagy at the cellular and organismal level is important for the development of therapeutic interventions against aging-related diseases.

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

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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

Similar content being viewed by others

References

  1. Feng Y, He D, Yao Z, Klionsky DJ (2014) The machinery of macroautophagy. Cell Res 24(1):24–41

    Article  CAS  PubMed  Google Scholar 

  2. Mizushima N, Komatsu M (2011) Autophagy: renovation of cells and tissues. Cell 147(4):728–741

    Article  CAS  Google Scholar 

  3. Lamark T, Johansen T (2012) Aggrephagy: selective disposal of protein aggregates by macroautophagy. Int J Cell Biol 2012:736905

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Kirkin V, Lamark T, Sou YS, Bjorkoy G, Nunn JL, Bruun JA, Shvets E, McEwan DG, Clausen TH, Wild P, Bilusic I, Theurillat JP, Overvatn A, Ishii T, Elazar Z, Komatsu M, Dikic I, Johansen T (2009) A role for NBR1 in autophagosomal degradation of ubiquitinated substrates. Mol Cell 33(4):505–516

    Article  PubMed  Google Scholar 

  5. Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H, Overvatn A, Bjorkoy G, Johansen T (2007) p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem 282(33):24131–24145

    Article  CAS  PubMed  Google Scholar 

  6. Stolz A, Ernst A, Dikic I (2014) Cargo recognition and trafficking in selective autophagy. Nat Cell Biol 16(6):495–501

    Article  CAS  PubMed  Google Scholar 

  7. Jain A, Rusten TE, Katheder N, Elvenes J, Bruun JA, Sjottem E, Lamark T, Johansen T (2015) p62/Sequestosome-1, autophagy-related gene 8, and autophagy in Drosophila are regulated by nuclear factor erythroid 2-related factor 2 (NRF2), independent of transcription factor TFEB. J Biol Chem 290(24):14945–14962

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Nezis IP, Simonsen A, Sagona AP, Finley K, Gaumer S, Contamine D, Rusten TE, Stenmark H, Brech A (2008) Ref(2)P, the Drosophila melanogaster homologue of mammalian p62, is required for the formation of protein aggregates in adult brain. J Cell Biol 180(6):1065–1071

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Martinez-Lopez N, Athonvarangkul D, Singh R (2015) Autophagy and aging. Adv Exp Med Biol 847:73–87

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Lystad AH, Simonsen A (2015) Assays to monitor aggrephagy. Methods 75:112–119

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work is supported by BBSRC grants BB/L006324/1 and BB/P007856/1 awarded to Dr. Ioannis P. Nezis.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ioannis P. Nezis .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media New York

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Jacomin, AC., Nezis, I.P. (2018). Assays to Monitor Aggrephagy in Drosophila Brain. In: Turksen, K. (eds) Autophagy in Differentiation and Tissue Maintenance. Methods in Molecular Biology, vol 1854. Humana Press, New York, NY. https://doi.org/10.1007/7651_2018_157

Download citation

  • DOI: https://doi.org/10.1007/7651_2018_157

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8747-4

  • Online ISBN: 978-1-4939-8748-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics