Vital staining for cell death identifies Atg9a-dependent necrosis in developmental bone formation in mouse.
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.
Department of Molecular and Cellular Biology, Research Institute of Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-2 Nakamichi, Higashinari-ku, Osaka 537-8511, Japan.|Laboratory of Molecular Genetics, Department of Medical Genetics, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Yusuke
Y
Imagawa
Department of Inflammation Biology, Institute for Enzyme Research, Tokushima University, 3-18-15 Kuramoto-cho, Tokushima 770-8503, Japan.|Laboratory of Host Defense, WPI Immunology Frontier Research Center, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Tatsuya
T
Saitoh
Department of Molecular and Cellular Biology, Research Institute of Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-2 Nakamichi, Higashinari-ku, Osaka 537-8511, Japan.|Laboratory of Molecular Genetics, Department of Medical Genetics, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Yoshihide
Y
Tsujimoto
D000818:Animals; D001343:Autophagy / Q000502:physiology*; D000071187:Autophagy-Related Protein 5 / Q000235:genetics / Q000378:metabolism; D000071183:Autophagy-Related Proteins / Q000235:genetics / Q000378:metabolism; D001842:Bone and Bones / Q000166:cytology / Q000473:pathology*; D002462:Cell Membrane / Q000473:pathology; D005260:Female; D007202:Indicators and Reagents / Q000737:chemistry; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D023421:Models, Animal; D009336:Necrosis / Q000473:pathology*; D010012:Osteogenesis / Q000502:physiology; D011419:Propidium / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods*; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism
C495773:Atg5 protein, mouse; C549015:Atg9A protein, mouse; D000071187:Autophagy-Related Protein 5; D000071183:Autophagy-Related Proteins; D007202:Indicators and Reagents; D008565:Membrane Proteins; D033921:Vesicular Transport Proteins; D011419:Propidium