Methods in renal research: Measurement of autophagic flux in the renal cortex ex vivo

Nephrology (Carlton). 2018 Sep;23(9):815-820. doi: 10.1111/nep.13251.

Abstract

The role of autophagy in the kidney and many nephrological diseases has gained prominence in recent years. Much of this research has been focused on markers of autophagy that are static and reveal little about the state of this dynamic pathway. Other mechanistic investigations are limited to in vitro studies, that often provide circumstantial evidence of autophagic flux. Here we describe a method for measuring autophagic flux ex vivo that allows more direct observations to be made in situ regarding the state of autophagic flux within the renal cortex of a single animal.

Keywords: autophagic flux; autophagosomes; autophagy; ex vivo; renal cortex.

MeSH terms

  • Animals
  • Autophagosomes / drug effects
  • Autophagosomes / metabolism
  • Autophagosomes / ultrastructure*
  • Autophagy* / drug effects
  • Autophagy-Related Proteins / genetics
  • Autophagy-Related Proteins / metabolism
  • Gene Expression Regulation
  • In Vitro Techniques
  • Kidney Cortex / drug effects
  • Kidney Cortex / metabolism
  • Kidney Cortex / ultrastructure*
  • Macrolides / pharmacology
  • Male
  • Mice, Inbred C57BL
  • Microscopy, Electron, Transmission*
  • Signal Transduction
  • Sirolimus / pharmacology
  • Time Factors

Substances

  • Autophagy-Related Proteins
  • Macrolides
  • bafilomycin A1
  • Sirolimus