Measuring Autophagy in Pancreatitis

Methods Mol Biol. 2019:1880:541-554. doi: 10.1007/978-1-4939-8873-0_35.

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.

Keywords: Autophagosomes; Caerulein; Ela1-VMP1 transgenic mouse; Pancreatitis; Selective autophagy.

MeSH terms

  • Acinar Cells
  • Animals
  • Autophagosomes / pathology*
  • Autophagosomes / ultrastructure
  • Autophagy*
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods
  • Cell Line
  • Ceruletide / toxicity
  • Disease Models, Animal
  • Enzyme Precursors / metabolism*
  • Humans
  • Lysosomes / metabolism
  • Male
  • Mice
  • Microscopy, Electron / instrumentation
  • Microscopy, Electron / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Pancreas / cytology
  • Pancreas / pathology*
  • Pancreatectomy
  • Pancreatitis / chemically induced
  • Pancreatitis / pathology*
  • Pancreatitis / surgery
  • Rats
  • Secretory Vesicles / pathology

Substances

  • Enzyme Precursors
  • Ceruletide