Abstract
Optic atrophy 1 ( OPA1 ) mutations cause dominant optic atrophy (DOA) with retinal ganglion cell (RGC) and optic nerve degeneration. The mechanism for the selective degeneration of RGCs in DOA remains elusive. To address the mechanism, we reduced OPA1 protein expression in cell lines and RGCs by RNA interference. OPA1 loss results in mitochondrial fragmentation, deficiency in oxidative phosphorylation, decreased ATP levels, decreased mitochondrial Ca 2+ retention capacity, reduced mtDNA copy numbers, and sensitization to apoptotic insults. We demonstrate profound cristae depletion and loss of crista junctions in OPA1 knockdown cells, whereas the remaining crista junctions preserve their normal size. OPA1-depleted cells exhibit decreased agonist-evoked mitochondrial Ca 2+ transients and corresponding reduction of NAD + to NADH, but the impairment in NADH oxidation leads to an overall more reduced mitochondrial NADH pool. Although in our model OPA1 loss in RGCs has no apparent impact on mitochondrial morphology, it decreases buffering of cytosolic Ca 2+ and sensitizes RGCs to excitotoxic injury. Exposure to glutamate triggers delayed calcium deregulation (DCD), often in a reversible manner, indicating partial resistance of RGCs to this injury. However, when OPA1 is depleted, DCD becomes irreversible. Thus, our data show that whereas OPA1 is required for mitochondrial fusion, maintenance of crista morphology and oxidative phosphorylation, loss of OPA1 also results in defective Ca 2+ homeostasis.
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Accession codes
Abbreviations
- DOA:
-
dominant optic atrophy
- DCD:
-
delayed Ca2+ deregulation
- Drp1:
-
dynamin-related protein 1
- [Ca2+]c:
-
cytoplasmic Ca2+ concentration
- [Ca2+]m:
-
mitochondrial Ca2+ concentration
- Δψm:
-
mitochondrial membrane potential
- FCCP:
-
carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone
- MPT:
-
mitochondrial permeability transition
- mtDNA:
-
mitochondrial DNA
- nDNA:
-
nuclear DNA
- OMM:
-
outer mitochondrial membrane
- RGC:
-
retinal ganglion cell
- siRNA:
-
small interfering RNA
- TPP+:
-
tetraphenylphosphonium
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Acknowledgements
We thank Dr. E Monosov for help in fluorescence imaging, Dr. B Barres and colleagues for teaching us to prepare RGC cultures, Dr. Y Kubo for OPA1 antibodies, Dr. A Andreyev for critical reading of the manuscript, and WD Lee for technical assistance. This work was supported by NIH Grants R01 NS047456, R01 NS047456, and R01 EY016164 (to EB-W), NIH Grant R01 EY018658 (to W-KJ), and NIH Grants P41RR004050 and R01 NS14718 (to MHE).
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Kushnareva, Y., Gerencser, A., Bossy, B. et al. Loss of OPA1 disturbs cellular calcium homeostasis and sensitizes for excitotoxicity. Cell Death Differ 20, 353–365 (2013). https://doi.org/10.1038/cdd.2012.128
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DOI: https://doi.org/10.1038/cdd.2012.128
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