Abstract
The ability of mitochondria to buffer a rapid rise in cytosolic Ca2+ is a hallmark of proper cell homeostasis. Here, we employed m-3M3FBS, a putative phospholipase C (PLC) agonist, to explore the relationships between intracellular Ca2+ imbalance, mitochondrial physiology, and cell death. m-3M3FBS induced a potent dose-dependent Ca2+ release from the endoplasmic reticulum (ER), followed by a rise in intra-mitochondrial Ca2+. When the latter exceeded the organelle buffering capacity, an abrupt mitochondrial inner membrane permeabilization (MIMP) occurred, releasing matrix contents into the cytosol. MIMP was followed by cell death that was independent of Bcl-2 family members and inhibitable by the intracellular Ca2+ chelator BAPTA-AM. Cyclosporin A (CsA), capable of blocking the mitochondrial permeability transition (MPT), completely prevented cell death induced by m-3M3FBS. However, CsA acted upstream of mitochondria by preventing Ca2+ release from ER stores. Therefore, loss of Ca2+ intracellular balance and mitochondrial Ca2+ overload followed by MIMP induced a cell death process that is distinct from Bcl-2 family-regulated mitochondrial outer membrane permeabilization (MOMP). Further, the inhibition of cell death by CsA or its analogues can be independent of effects on the MPT.
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Acknowledgements
We thank Dr. Tudor Moldoveanu for providing MEFs plcγ1−/− and plcγ2−/−. Transmission electron microscopy images were acquired at the Cell & Tissue Imaging Center of St. Jude Children’s Research Hospital. G.Q. thanks Dr. Tudor Moldoveanu and Prof. Nazzareno Capitanio for a critical review of the manuscript. Cartoons were created with BioRender.com.
Funding
This research was supported by R35CA231620 from the U.S. National Cancer Institute. Transmission electron microscopy images acquisition was supported by St. Jude Children’s Research Hospital and NCI P30 CA021765. Center for Advanced Genome Engineering (CAGE) of St. Jude Children’s Research Hospital was supported by Cancer Center Grant P30-CA021765. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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GQ, FL, CSG, MA, HS and SN performed the experiments and analyzed the results. SMP-M, JM, JP, ZR and DRG supervised the study and provided access to material and facilities. GQ, FL and DRG conceived and designed the work. GQ and DRG wrote the manuscript. All the authors reviewed the manuscript.
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Quarato, G., Llambi, F., Guy, C.S. et al. Ca2+-mediated mitochondrial inner membrane permeabilization induces cell death independently of Bax and Bak. Cell Death Differ 29, 1318–1334 (2022). https://doi.org/10.1038/s41418-022-01025-9
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DOI: https://doi.org/10.1038/s41418-022-01025-9
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