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Voltage-dependent anion channels are dispensable for mitochondrial-dependent cell death

Abstract

Mitochondria are critically involved in necrotic cell death induced by Ca2+ overload, hypoxia and oxidative damage. The mitochondrial permeability transition (MPT) pore — a protein complex that spans both the outer and inner mitochondrial membranes — is considered the mediator of this event and has been hypothesized to minimally consist of the voltage-dependent anion channel (Vdac) in the outer membrane, the adenine-nucleotide translocase (Ant) in the inner membrane and cyclophilin-D in the matrix1,2,3. Here, we report the effects of deletion of the three mammalian Vdac genes on mitochondrial-dependent cell death. Mitochondria from Vdac1-, Vdac3-, and Vdac1Vdac3-null mice exhibited a Ca2+- and oxidative stress-induced MPT that was indistinguishable from wild-type mitochondria. Similarly, Ca2+- and oxidative-stress-induced MPT and cell death was unaltered, or even exacerbated, in fibroblasts lacking Vdac1, Vdac2, Vdac3, Vdac1–Vdac3 and Vdac1–Vdac2–Vdac3. Wild-type and Vdac-deficient mitochondria and cells also exhibited equivalent cytochrome c release, caspase cleavage and cell death in response to the pro-death Bcl-2 family members Bax and Bid. These results indicate that Vdacs are dispensable for both MPT and Bcl-2 family member-driven cell death.

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Figure 1: Mitochondrial permeability transition in Vdac1- and Vdac3-deficient mitochondria.
Figure 2: Cytochrome c release induced by Ca2+, oxidative stress, Bax and tBid.
Figure 3: Mitochondrial permeability transition and cell death in Vdac1- and Vdac3-null MEFs.
Figure 4: Cell death in Vdac-deficient MEFs.
Figure 5: MPT and cell death in Vdac2-null fibroblasts and in MEFs deficient in Vdac proteins.

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Acknowledgements

We are very grateful to B. Fang for the generous gift of the Bax adenovirus, R. Gottlieb for recombinant Bax and tBid, and the late S. Korsmeyer for Bax–Bak gene-targeted mice. This work was supported by grants from the National Institutes of Health (J.D.M. and W.J.C.), an American Heart Association Scientist Development Grant (C.P.B.) and Established Investigator Grant (J.D.M.), a National Institutes of Health NRSA award (R.A.K.), The Children's Hospital Research Foundation (C.P.B.), and the Fondation Leducq (Heart failure network grant to J.D.M).

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C.P.B. and J.D.M. conceived the research project, C.P.B. performed all experimentation except the ischaemia–reperfusion experiment, which was performed by R.A.K. T.S. and W.J.C. provided critical reagents and valuable discussion.

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Correspondence to Jeffery D. Molkentin.

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The authors declare no competing financial interests.

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Supplementary Figures S1, S2, S3, S4 and S5 (PDF 663 kb)

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Baines, C., Kaiser, R., Sheiko, T. et al. Voltage-dependent anion channels are dispensable for mitochondrial-dependent cell death. Nat Cell Biol 9, 550–555 (2007). https://doi.org/10.1038/ncb1575

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