Advertisement

Autophagy pp 541-554 | Cite as

Measuring Autophagy in Pancreatitis

  • Alejandro Ropolo
  • Daniel Grasso
  • Maria I. VaccaroEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1880)

Abstract

Acute pancreatitis is one of the first pathological processes where autophagy has been described in a human tissue. Autophagy, autodigestion, and cell death are early cellular events in acute pancreatitis. Recent advances in understanding autophagy highlight its importance in pathological conditions. However, methods for monitoring autophagic activity during complex diseases, involving highly differentiated secretory cells, are complicated, and the results are sometimes misinterpreted. Here, we describe methods used to identify autophagic structures and to measure autophagic flux in cultured cell models and animal models of pancreatitis. We also briefly describe the pancreas specific autophagy mouse model that was useful to understand the actual role of autophagy in pancreatitis and to identify a novel selective autophagy pathway named zymophagy. Lastly, we describe the immunomagnetic isolation of autophagosomes and the detection of autophagy in pancreatic tissue samples derived from humans.

Key words

Pancreatitis Caerulein Selective autophagy Autophagosomes Ela1-VMP1 transgenic mouse 

References

  1. 1.
    Sah RP, Saluja A (2011) Molecular mechanisms of pancreatic injury. Curr Opin Gastroenterol 27:444–451CrossRefGoogle Scholar
  2. 2.
    Lankisch PG, Apte M, Banks PA (2015) Acute Pancreatitis. Lancet 386:85–96CrossRefGoogle Scholar
  3. 3.
    Helin H, Mero M, Markkula H, Helin M (1980) Pancreatic acinar ultrastructure in human acute pancreatitis. Virchows Arch 387:259–270CrossRefGoogle Scholar
  4. 4.
    Gorelick FS, Adler G, Kern HF et al (1993) Cerulein-induced pancreatitis. In: Go VLW, DiMagno EP, Gardner JD et al (eds) The pancreas: biology, pathophysiology and disease, 2nd edn. Raven Press, New York, NY, pp 501–526Google Scholar
  5. 5.
    Chandra R, Liddle RA (2015) Regulation of pancreatic secretion. Pancreapedia.  https://doi.org/10.3998/panc.2015.38
  6. 6.
    Grasso D, Ropolo A, Ré L et al (2011) Zymophagy, a novel selective autophagy pathway mediated by VMP1-USP9x-p62, prevents pancreatic cell death. J Biol Chem 286:8308–8324CrossRefGoogle Scholar
  7. 7.
    Vaccaro MI (2012) Zymophagy: selective autophagy of secretory granules. Int J Cell Biol 2012:396705.  https://doi.org/10.1155/2012/396705CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Williams JA (2008) Receptor-mediated signal transduction pathways and the regulation of pancreatic acinar cell function. Curr Opin Gastroenterol 24:573–579CrossRefGoogle Scholar
  9. 9.
    Niederau C, Ferrell LD, Grendell JH (1985) Caerulein-induced acute necrotizing pancreatitis in mice: protective effects of proglumide, benzotript, and secretin. Gastroenterology 88:1192–1204CrossRefGoogle Scholar
  10. 10.
    Gorelick FS, Lerch MM (2017) Do animal models of acute pancreatitis reproduce human disease? Cell Mol Gastroenterol Hepatol 4:251–262CrossRefGoogle Scholar
  11. 11.
    Klionsky D et al (2016) Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy 12:1–222CrossRefGoogle Scholar
  12. 12.
    Ropolo A, Grasso D, Pardo R et al (2007) The pancreatitis-induced vacuole membrane protein 1 triggers autophagy in mammalian cells. J Biol Chem 282:37124–37133CrossRefGoogle Scholar
  13. 13.
    Vaccaro MI, Grasso D, Ropolo A, Iovanna JL, Cerquetti MC (2003) VMP1 expression correlates with acinar cell cytoplasmic vacuolization in arginine-induced acute pancreatitis. Pancreatology 3:69–74CrossRefGoogle Scholar
  14. 14.
    Vaccaro MI (2008) Autophagy and pancreas disease. Pancreatology 8:425–429CrossRefGoogle Scholar
  15. 15.
    Molejon MI, Ropolo A, Vaccaro MI (2013) VMP1 is a new player in the regulation of the autophagy-specific phosphatidylinositol 3-kinase complex activation. Autophagy 9:933–935CrossRefGoogle Scholar
  16. 16.
    Molejon MI, Ropolo A, Re AL, Boggio V, Vaccaro MI (2013) The VMP1-Beclin 1 interaction regulates autophagy induction. Sci Rep 3:1055.  https://doi.org/10.1038/srep01055CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Nascimbeni AC, Giordano F, Dupont N, Grasso D, Vaccaro MI, Codogno P, Morel E (2017) ER-plasma membrane contact sites contribute to autophagosome biogenesis by regulation of local PI3P synthesis. EMBO J 36:2018–2033CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Alejandro Ropolo
    • 1
  • Daniel Grasso
    • 1
  • Maria I. Vaccaro
    • 1
    Email author
  1. 1.Pathophysiology Department, Institute of Biochemistry and Molecular Medicine (CONICET), School of Pharmacy and BiochemistryUniversity of Buenos AiresBuenos AiresArgentina

Personalised recommendations