Vital staining for cell death identifies Atg9a-dependent necrosis in developmental bone formation in mouse

Nat Commun. 2016 Nov 4:7:13391. doi: 10.1038/ncomms13391.

Abstract

Programmed cell death has a crucial role in various biological events, including developmental morphogenesis. Recent evidence indicates that necrosis contributes to programmed cell death in addition to apoptosis, but it is unclear whether necrosis acts as a compensatory mechanism for failure of apoptosis or has an intrinsic role during development. In contrast to apoptosis, there have been no techniques for imaging physiological necrosis in vivo. Here we employ vital staining using propidium iodide to identify cells with plasma membrane disruption (necrotic cells) in mouse embryos. We discover a form of necrosis at the bone surface, which does not occur in embryos with deficiency of the autophagy-related gene Atg9a, although it is unaffected by Atg5 knockout. We also find abnormalities of the bone surface in Atg9a knockout mice, suggesting an important role of Atg9a-dependent necrosis in bone surface formation. These findings suggest that necrosis has an active role in developmental morphogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autophagy / physiology*
  • Autophagy-Related Protein 5 / genetics
  • Autophagy-Related Protein 5 / metabolism
  • Autophagy-Related Proteins / genetics
  • Autophagy-Related Proteins / metabolism
  • Bone and Bones / cytology
  • Bone and Bones / pathology*
  • Cell Membrane / pathology
  • Female
  • Indicators and Reagents / chemistry
  • Male
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Models, Animal
  • Necrosis / pathology*
  • Osteogenesis / physiology
  • Propidium / chemistry*
  • Staining and Labeling / methods*
  • Vesicular Transport Proteins / genetics
  • Vesicular Transport Proteins / metabolism

Substances

  • Atg5 protein, mouse
  • Atg9A protein, mouse
  • Autophagy-Related Protein 5
  • Autophagy-Related Proteins
  • Indicators and Reagents
  • Membrane Proteins
  • Vesicular Transport Proteins
  • Propidium