TY - JOUR
T1 - Unique spectral signatures of the nucleic acid dye acridine orange can distinguish cell death by apoptosis and necroptosis
AU - Plemel, Jason R.
AU - Caprariello, Andrew V.
AU - Keough, Michael B.
AU - Henry, Tyler J.
AU - Tsutsui, Shigeki
AU - Chu, Tak H.
AU - Schenk, Geert J.
AU - Klaver, Roel
AU - Wee Yong, V.
AU - Stys, Peter K.
PY - 2017/4/1
Y1 - 2017/4/1
N2 - Cellular injury and death are ubiquitous features of disease, yet tools to detect them are limited and insensitive to subtle pathological changes. Acridine orange (AO), a nucleic acid dye with unique spectral properties, enables real-time measurement of RNA and DNA as proxies for cell viability during exposure to various noxious stimuli. This tool illuminates spectral signatures unique to various modes of cell death, such as cells undergoing apoptosis versus necrosis/ necroptosis. This new approach also shows that cellular RNA decreases during necrotic, necroptotic, and apoptotic cell death caused by demyelinating, ischemic, and traumatic injuries, implying its involvement in a wide spectrum of tissue pathologies. Furthermore, cells with pathologically low levels of cytoplasmic RNA are detected earlier and in higher numbers than with standard markers including TdT-mediated dUTP biotin nick-end labeling and cleaved caspase 3 immunofluorescence. Our technique highlights AO-labeled cytoplasmic RNA as an important early marker of cellular injury and a sensitive indicator of various modes of cell death in a range of experimental models.
AB - Cellular injury and death are ubiquitous features of disease, yet tools to detect them are limited and insensitive to subtle pathological changes. Acridine orange (AO), a nucleic acid dye with unique spectral properties, enables real-time measurement of RNA and DNA as proxies for cell viability during exposure to various noxious stimuli. This tool illuminates spectral signatures unique to various modes of cell death, such as cells undergoing apoptosis versus necrosis/ necroptosis. This new approach also shows that cellular RNA decreases during necrotic, necroptotic, and apoptotic cell death caused by demyelinating, ischemic, and traumatic injuries, implying its involvement in a wide spectrum of tissue pathologies. Furthermore, cells with pathologically low levels of cytoplasmic RNA are detected earlier and in higher numbers than with standard markers including TdT-mediated dUTP biotin nick-end labeling and cleaved caspase 3 immunofluorescence. Our technique highlights AO-labeled cytoplasmic RNA as an important early marker of cellular injury and a sensitive indicator of various modes of cell death in a range of experimental models.
UR - http://www.scopus.com/inward/record.url?scp=85021848520&partnerID=8YFLogxK
U2 - https://doi.org/10.1083/jcb.201602028
DO - https://doi.org/10.1083/jcb.201602028
M3 - Article
C2 - 28264914
SN - 0021-9525
VL - 216
SP - 1163
EP - 1181
JO - Journal of cell biology
JF - Journal of cell biology
IS - 4
ER -