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

• Published in: Nature communications 2016-11, Vol.7 (1), p.13391-13391, Article 13391
• Main Authors: Imagawa, Yusuke, Saitoh, Tatsuya, Tsujimoto, Yoshihide
• Format: Article
• Language: English
• Subjects: 631/136/818; 631/80/82; 64/110; 82/51; 96/106; 96/2; 96/34; 96/63; 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 death; Cell Membrane - pathology; Female; Humanities and Social Sciences; Indicators and Reagents - chemistry; Male; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice; Mice, Inbred C57BL; Mice, Knockout; Models, Animal; multidisciplinary; Necrosis - pathology; Osteogenesis - physiology; Propidium - chemistry; Science; Science (multidisciplinary); Staining and Labeling - methods; Vesicular Transport Proteins - genetics; Vesicular Transport Proteins - metabolism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms13391

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. Apoptosis occurs in numerous developmental processes but a role for necrosis in development is unclear. Here, the authors develop a detecting system of necrosis in the developing mouse and find a form of necrosis that is dependent on the autophagy-related gene Atg9a .