Flannery 2015 Abstract MiP2015

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Quantitative live-cell imaging of mitochondrial network morphology in neurodegenerative conditions.

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Flannery PJ, Sitarz K, Yu-Wai-Man P (2015)

Event: MiP2015

Mitochondrial dysfunction is a well-established hallmark of aging and neurodegenerative diseases. Maintenance of mitochondrial dynamics is essential for mitochondrial health maintenance and disturbances in mitochondrial dynamics have been implicated in a number of neurodegenerative processes. Moreover, patients with mutations in mitochondrial proteins involved in mitochondrial fusion, namely, Mfn2 and OPA1 genes have been associated with Charcot-Marie-Tooth disease type 2A, hereditary motor and sensory neuropathy VI, and autosomal optic atrophy (ADOA), respectively. In addition to its major role in mitochondrial fusion, OPA1 is an inner mitochondrial membrane protein which is involved in apoptosis, cristae structure, mtDNA replication maintenance and mitochondrial potential, all potential hallmarks of neurodegenerative conditions [1].

To study the role of mitochondrial fusion in neurodegenerative processes, we have adapted qualitative methods based on subjective classification of organelle morphology into defined categories to a quantitative protocol which uses mitotracker staining of the mitochondrial network followed by live-cell confocal imaging combined with Huygens Essential for deconvolution and image analysis. The output allows a quantitative assessment of mitochondrial length and volume in different physiological and disease conditions [2,3]. Using this technique, we have successfully imaged the mitochondrial network with a high degree of cell-to- cell reproducibility in both control fibroblasts and patient-derived primary fibroblasts with carrying pathogenic OPA1 mutations. OPA1-mutant fibroblasts showed clear morphological changes when compared with control fibroblasts under both basal and mitochondrial OXPHOS stress conditions.

Our study demonstrates the advantages of in-depth quantitative analysis of mitochondrial network morphology by using a reproducible protocol that is applicable to a wide range of neurodegenerative diseases.


Labels: MiParea: Instruments;methods, mt-Structure;fission;fusion, mt-Membrane, mtDNA;mt-genetics, mt-Medicine  Pathology: Other  Stress:Mitochondrial disease 

Tissue;cell: Fibroblast 




Event: A1, Poster, P-flash  MiP2015 

Affiliations

Wellcome Trust Centre for Mitochondrial research, Institute of Genetic Research, Newcastle University; Newcastle upon Tyne, UK. - p.f.flannery@ncl.ac.uk

References and acknowledgements

  1. Burte F, Carelli V, Chinnery PF (2015) Disturbed mitochondrial dynamics and neurodegenerative disorders. Nat Rev Neurol 11:11-24.
  2. Zanna C, Ghelli A, Karbowski M, Youle RJ, Schimpf S (2008) OPA1 mutations associated with dominant optic atrophy impair oxidative phosphorylation and mitochondrial fusion. Brain 131:352-67.
  3. Kao Shu-Huei, Yen May-Yung, Wang An-Guor (2015) Changes in mitochondrial morphology and bioenergetics in human lymphoblastoid cells with four novel OPA1 mutations. Investigative Ophthalmology and Visual Science 56:4:2269-78.

Supported by the Medical Research Council (MRC, UK)