Abstract
Using confocal microscopy, we investigated mechanisms underlying loss of plasma membrane integrity during necrotic death of cultured hepatic sinusoidal endothelial cells exposed to 2.5 mM potassium cyanide (chemical hypoxia). After 2–3 h, the anionic fluorophore calcein abruptly began to enter the cytosol, and nuclei labeled with cationic propidium after another 2–5 min. As calcein permeated, growth of blebs on the plasma membrane accelerated. Lucifer yellow, another anionic fluorophore, entered identically to calcein, whereas high molecular weight dextrans (40–2000 kDa) entered like propidium. Glycine slowed, but did not prevent calcein entry, whereas permeation of propidium and high molecular weight dextrans was blocked completely by glycine. These findings suggest that opening of a glycine-sensitive organic anion channel, or death channel, precipitates a metastable state characterized by rapid cell swelling and bleb growth. This metastable state culminates in non-specific breakdown of the plasma membrane permeability barrier and irreversible cell death. Cell Death and Differentiation (2001) 8, 850–858
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Abbreviations
- KRH:
-
Krebs-Ringer-HEPES buffer
- MDCK:
-
Madin-Darby canine kidney cells
- N.A.:
-
numerical aperture
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Acknowledgements
This work was supported, in part, by Grants DK37034, AG13637 and AA11605 from the National Institutes of Health. Core imaging facilities were supported, in part, by 1-P50-AA11605 and 5-P30-DK34987 to the Center for Alcohol Studies and the Center for Gastrointestinal Biology and Disease. Portions of this work were presented at the 47th Annual Meeting of the American Association for the Study of Liver Diseases, Chicago, IL, USA, November 8–12, 1996.
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Nishimura, Y., Lemasters, J. Glycine blocks opening of a death channel in cultured hepatic sinusoidal endothelial cells during chemical hypoxia. Cell Death Differ 8, 850–858 (2001). https://doi.org/10.1038/sj.cdd.4400877
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DOI: https://doi.org/10.1038/sj.cdd.4400877
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