Assays to Monitor Mitophagy in Drosophila

Methods Mol Biol. 2019:1880:643-653. doi: 10.1007/978-1-4939-8873-0_42.

Abstract

Autophagy is a central pathway utilized by many eukaryotic cells in order to recycle intracellular constituents, particularly under periods of nutrient scarcity or cellular damage. The process is evolutionarily conserved from yeast to mammals and can be highly selective with regard to the contents that are targeted for degradation. The availability of Drosophila transgenic lines and fluorophore-labeled autophagic markers allows nowadays for the more effortless visualization of the process within cells. Herein, we provide two protocols to prepare Drosophila samples for confocal and transmission electron microscopy for in vivo monitoring of mitophagy, a specific type of autophagy for the clearance of damaged or superfluous mitochondria from cells.

Keywords: Autophagy; Complex V; Confocal microscopy; Drosophila; Electron microscopy; Fat body; Mitophagy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Biological Assay / instrumentation
  • Biological Assay / methods*
  • Drosophila / physiology
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism
  • Fat Body / metabolism
  • Female
  • Fluorescent Dyes / chemistry
  • Intravital Microscopy / instrumentation
  • Intravital Microscopy / methods*
  • Larva / physiology
  • Male
  • Microscopy, Confocal / instrumentation
  • Microscopy, Confocal / methods
  • Microscopy, Electron, Transmission / instrumentation
  • Microscopy, Electron, Transmission / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Mitochondria / metabolism
  • Mitophagy / physiology*
  • Models, Animal

Substances

  • Atg8a protein, Drosophila
  • Drosophila Proteins
  • Fluorescent Dyes