Abstract
The goal of the current study was to determine the roles of ATP content, endoplasmic reticulum (ER) Ca2+ stores, cytosolic free Ca2+ (Ca2+f) and calpain activity in the signaling of rabbit renal proximal tubular (RPT) cell death (oncosis). Increasing concentrations (0.3–10 μM) of the mitochondrial inhibitor antimycin A produced rapid ATP depletion that correlated to a rapid and sustained increase in Ca2+f, but not phospholipase C activation. The ER Ca2+-ATPase inhibitors thapsigargin (5 μM) or cyclopiazonic acid (100 μM) alone produced similar but transient increases in Ca2+f. Pretreatment with thapsigargin prevented antimycin A-induced increases in Ca2+f and antimycin A pretreatment prevented thapsigargin-induced increases in Ca2+f. Calpain activity increased in conjunction with ER Ca2+ release. Pretreatment, but not post-treatment, with thapsigargin or cyclopiazonic acid prevented antimycin A-induced cell death. These data demonstrate that extensive ATP depletion signals oncosis through ER Ca2+ release, a sustained increase in Ca2+f and calpain activation. Depletion of ER Ca2+ stores prior to toxicant exposure prevents increases in Ca2+f and oncosis.
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Abbreviations
- ER:
-
endoplasmic reticulum
- Ca2+f:
-
cytosolic free Ca2+; RPT, renal proximal tubule
- SERCA:
-
smooth endoplasmic reticulum Ca2+-ATPase; LDH, lactate dehydrogenase
- IP3:
-
inositiol trisphosphate
- PLC:
-
phospholipase C
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Acknowledgements
We would like to thank Dr. Phil Mayeux for the use of the spectrofluorophotometer during this study. We would also like to thank Dr. Brian Cummings for his review of the manuscript. This work was supported in part by NIH ES-09129 (RG Schnellmann) and American Heart Association/Heartland Affiliate predoctoral fellowships to JF Harriman and X Liu.
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Harriman, J., Liu, X., Aleo, M. et al. Endoplasmic reticulum Ca2+ signaling and calpains mediate renal cell death. Cell Death Differ 9, 734–741 (2002). https://doi.org/10.1038/sj.cdd.4401029
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DOI: https://doi.org/10.1038/sj.cdd.4401029
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