Nonviability of cells with oxidative defects in galactose medium: A screening test for affected patient fibroblasts

Robinson, Brian H.; Petrova‐Benedict, Roumyana; Buncic, J. Raymond; Wallace, Douglas C.

doi:10.1016/0885-4505(92)90056-5

Cited by 249 publications

(245 citation statements)

References 11 publications

Supporting

Mentioning

231

Contrasting

Unclassified

Order By: Relevance

“…This finding was in agreement with previous studies showing that cultured cells with defective mitochondrial metabolism often die in glucosefree galactose medium (20,21). Indeed, in glucose medium, most of the total ATP yield of proliferating cells is produced by glycolytic glucose breakdown to lactate, and only a minimal amount of pyruvate is oxidized to CO 2 and water.…”

supporting

confidence: 93%

“…Under this experimental condition, the reduced rate of the glycolytic pathway causes the forced oxidation of pyruvate through the mitochondrial respiratory chain. This experimental condition has been used previously to identify an impaired respiratory function (20,21), and Hofhaus et al (19) did report a reduced ability of LHON/11778 cybrids to grow in galactose medium (19). Our study extended this seminal observation to the whole range of the main LHON pathogenic mutations, also showing that growth impairment was somewhat milder in 11778 cybrids as compared with those carrying the 3460 and 14484 mutations.…”

Section: Galactose Medium Induces Cell Death In Lhon Cybrids-supporting

confidence: 72%

See 1 more Smart Citation

Leber's Hereditary Optic Neuropathy (LHON) Pathogenic Mutations Induce Mitochondrial-dependent Apoptotic Death in Transmitochondrial Cells Incubated with Galactose Medium

Ghelli

Zanna

Porcelli

et al. 2003

Journal of Biological Chemistry

172

128

View full text Add to dashboard Cite

Leber's hereditary optic neuropathy (LHON), a maternally inherited form of central vision loss, is associated with mitochondrial DNA pathogenic point mutations affecting different subunits of complex I. We here report that osteosarcoma-derived cytoplasmic hybrids (cybrid) cell lines harboring one of the three most frequent LHON pathogenic mutations, at positions 11778/ND4, 3460/ND1, and 14484/ND6, undergo cell death when galactose replaces glucose in the medium, contrary to control cybrids that maintain some growth capabilities. This is a well known way to produce a metabolic stress, forcing the cells to rely on the mitochondrial respiratory chain to produce ATP. We demonstrate that LHON cybrid cell death is apoptotic, showing chromatin condensation and nuclear DNA laddering. Moreover, we also document the mitochondrial involvement in the activation of the apoptotic cascade, as shown by the increased release of cytochrome c into the cytosol in LHON cybrid cells as compared with controls. Cybrids bearing the 3460/ND1 and 14484/ND6 mutations seemed more readily prone to undergo apoptosis as compared with the 11778/ ND4 mutation. In conclusion, LHON cybrid cells forced by the reduced rate of glycolytic flux to utilize oxidative metabolism are sensitized to an apoptotic death through a mechanism involving mitochondria.

show abstract

supporting

confidence: 93%

Section: Galactose Medium Induces Cell Death In Lhon Cybrids-supporting

confidence: 72%

Leber's Hereditary Optic Neuropathy (LHON) Pathogenic Mutations Induce Mitochondrial-dependent Apoptotic Death in Transmitochondrial Cells Incubated with Galactose Medium

Ghelli

Zanna

Porcelli

et al. 2003

Journal of Biological Chemistry

172

128

View full text Add to dashboard Cite

show abstract

“…Mitochondrial network morphology was investigated in fibroblasts bearing the p.G488R and the p.A495V OPA1 mutations, after 48 hours incubation in galactose medium, a well‐established condition that forces cells to rely on OXPHOS for ATP synthesis 35. Whereas control fibroblasts maintained a mitochondrial filamentous network, mutant fibroblasts showed marked mitochondrial fragmentation (Fig 6A).…”

Section: Resultsmentioning

confidence: 99%

Syndromic parkinsonism and dementia associated with OPA1 missense mutations

Carelli

Musumeci

Caporali

et al. 2015

Annals of Neurology

160

106

View full text Add to dashboard Cite

ObjectiveMounting evidence links neurodegenerative disorders such as Parkinson disease and Alzheimer disease with mitochondrial dysfunction, and recent emphasis has focused on mitochondrial dynamics and quality control. Mitochondrial dynamics and mtDNA maintenance is another link recently emerged, implicating mutations in the mitochondrial fusion genes OPA1 and MFN2 in the pathogenesis of multisystem syndromes characterized by neurodegeneration and accumulation of mtDNA multiple deletions in postmitotic tissues. Here, we report 2 Italian families affected by dominant chronic progressive external ophthalmoplegia (CPEO) complicated by parkinsonism and dementia.MethodsPatients were extensively studied by optical coherence tomography (OCT) to assess retinal nerve fibers, and underwent muscle and brain magnetic resonance spectroscopy (MRS), and muscle biopsy and fibroblasts were analyzed. Candidate genes were sequenced, and mtDNA was analyzed for rearrangements.ResultsAffected individuals displayed a slowly progressive syndrome characterized by CPEO, mitochondrial myopathy, sensorineural deafness, peripheral neuropathy, parkinsonism, and/or cognitive impairment, in most cases without visual complains, but with subclinical loss of retinal nerve fibers at OCT. Muscle biopsies showed cytochrome c oxidase‐negative fibers and mtDNA multiple deletions, and MRS displayed defective oxidative metabolism in muscle and brain. We found 2 heterozygous OPA1 missense mutations affecting highly conserved amino acid positions (p.G488R, p.A495V) in the guanosine triphosphatase domain, each segregating with affected individuals. Fibroblast studies showed a reduced amount of OPA1 protein with normal mRNA expression, fragmented mitochondria, impaired bioenergetics, increased autophagy and mitophagy.InterpretationThe association of CPEO and parkinsonism/dementia with subclinical optic neuropathy widens the phenotypic spectrum of OPA1 mutations, highlighting the association of defective mitochondrial dynamics, mtDNA multiple deletions, and altered mitophagy with parkinsonism. Ann Neurol 2015;78:21–38

show abstract

“…Therefore, growth in galactose-containing medium is a sensitive means for identifying the defects in oxidative phosphorylation. 21 To test whether Ad.1891 infection produced an improvement in oxidative phosphorylation, the growth of the parental JCP239 cells in galactosecontaining medium was compared to that of 2-50 cells (JCP239 cells, which underwent two rounds of Ad.1891 infection with MOI ¼ 50). The growth rates of JCP239 and 2-50 cells were indistinguishable in glucose-containing medium (Figure 4a).…”

Section: Ad1891 Infection Improves the Biochemical Parameters Compromentioning

confidence: 99%

Selective elimination of mutant mitochondrial genomes as therapeutic strategy for the treatment of NARP and MILS syndromes

et al. 2008

View full text Add to dashboard Cite

Mitochondrial diseases are not uncommon, and may result from mutations in both nuclear and mitochondrial DNA (mtDNA). At present, only palliative therapies are available for these disorders, and interest in the development of efficient treatment protocols is high. Here, we demonstrate that in cells heteroplasmic for the T8993G mutation, which is a cause for the NARP and MILS syndromes, infection with an adenovirus, which encodes the mitochondrially targeted R.XmaI restriction endonuclease, leads to selective destruction of mutant mtDNA. This destruction proceeds in a timeand dose-dependent manner and results in cells with significantly increased rates of oxygen consumption and ATP production. The delivery of R.XmaI to mitochondria is accompanied by improvement in the ability to utilize galactose as the sole carbon source, which is a surrogate indicator of the proficiency of oxidative phosphorylation. Concurrently, the rate of lactic acid production by these cells, which is a marker of mitochondrial dysfunction, decreases. We further demonstrate that levels of phosphorylated P53 and gH2ax proteins, markers of nuclear DNA damage, do not change in response to infection with recombinant adenovirus indicating the absence of nuclear DNA damage and the relative safety of the technique. Finally, some advantages and limitations of the proposed approach are discussed.

show abstract

Nonviability of cells with oxidative defects in galactose medium: A screening test for affected patient fibroblasts

Cited by 249 publications

References 11 publications

Leber's Hereditary Optic Neuropathy (LHON) Pathogenic Mutations Induce Mitochondrial-dependent Apoptotic Death in Transmitochondrial Cells Incubated with Galactose Medium

Leber's Hereditary Optic Neuropathy (LHON) Pathogenic Mutations Induce Mitochondrial-dependent Apoptotic Death in Transmitochondrial Cells Incubated with Galactose Medium

Syndromic parkinsonism and dementia associated with OPA1 missense mutations

Selective elimination of mutant mitochondrial genomes as therapeutic strategy for the treatment of NARP and MILS syndromes

Contact Info

Product

Resources

About