Unique spectral signatures of the nucleic acid dye acridine orange can distinguish cell death by apoptosis and necroptosis

Jason R. Plemel, Andrew V. Caprariello, Michael B. Keough, Tyler J. Henry, Shigeki Tsutsui, Tak H. Chu, Geert J. Schenk, Roel Klaver, V. Wee Yong, Peter K. Stys*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)1163-1181
Number of pages19
JournalJournal of Cell Biology
Volume216
Issue number4
DOIs
Publication statusPublished - 1 Apr 2017

Cite this

Plemel, J. R., Caprariello, A. V., Keough, M. B., Henry, T. J., Tsutsui, S., Chu, T. H., ... Stys, P. K. (2017). Unique spectral signatures of the nucleic acid dye acridine orange can distinguish cell death by apoptosis and necroptosis. Journal of Cell Biology, 216(4), 1163-1181. https://doi.org/10.1083/jcb.201602028