title: Unique quadruple immunofluorescence assay demonstrates mitochondrial respiratory chain dysfunction in osteoblasts of aged and PolgA(-/-) mice.
abstract: Fragility fractures caused by osteoporosis affect millions of people worldwide every year with significant levels of associated morbidity, mortality and costs to the healthcare economy. The pathogenesis of declining bone mineral density is poorly understood but it is inherently related to increasing age. Growing evidence in recent years, especially that provided by mouse models, suggest that accumulating somatic mitochondrial DNA mutations may cause the phenotypic changes associated with the ageing process including osteoporosis. Methods to study mitochondrial abnormalities in individual osteoblasts, osteoclasts and osteocytes are limited and impair our ability to assess the changes seen with age and in animal models of ageing. To enable the assessment of mitochondrial protein levels, we have developed a quadruple immunofluorescence method to accurately quantify the presence of mitochondrial respiratory chain components within individual bone cells. We have applied this technique to a well-established mouse model of ageing and osteoporosis and show respiratory chain deficiency.
authors.affiliation: Wellcome Trust Centre for Mitochondrial Research, Institute for Neuroscience, Medical School, Newcastle University, United Kingdom.
authors.forename: Philip F
authors.initials: PF
authors.lastname: Dobson
authors.affiliation: Wellcome Trust Centre for Mitochondrial Research, Institute for Neuroscience, Medical School, Newcastle University, United Kingdom.
authors.forename: Mariana C
authors.initials: MC
authors.lastname: Rocha
authors.affiliation: Wellcome Trust Centre for Mitochondrial Research, Institute for Neuroscience, Medical School, Newcastle University, United Kingdom.
authors.forename: John P
authors.initials: JP
authors.lastname: Grady
authors.affiliation: Wellcome Trust Centre for Mitochondrial Research, Institute for Neuroscience, Medical School, Newcastle University, United Kingdom.
authors.forename: Alexia
authors.initials: A
authors.lastname: Chrysostomou
authors.affiliation: Wellcome Trust Centre for Mitochondrial Research, Institute for Neuroscience, Medical School, Newcastle University, United Kingdom.
authors.forename: Daniel
authors.initials: D
authors.lastname: Hipps
authors.affiliation: Musculoskeletal Research Group, Medical School, Newcastle University, United Kingdom.
authors.forename: Sharon
authors.initials: S
authors.lastname: Watson
authors.affiliation: Wellcome Trust Centre for Mitochondrial Research, Institute for Neuroscience, Medical School, Newcastle University, United Kingdom.|MRC/BBSRC Centre for Ageing and Vitality, Newcastle University, United Kingdom.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Greaves
authors.affiliation: Institute of Cellular Medicine, Newcastle University, United Kingdom.
authors.forename: David J
authors.initials: DJ
authors.lastname: Deehan
authors.affiliation: Wellcome Trust Centre for Mitochondrial Research, Institute for Neuroscience, Medical School, Newcastle University, United Kingdom.|MRC/BBSRC Centre for Ageing and Vitality, Newcastle University, United Kingdom.
authors.forename: Doug M
authors.initials: DM
authors.lastname: Turnbull
mesh_terms: D000375:Aging / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000074002:DNA Polymerase gamma / Q000235:genetics*; D004195:Disease Models, Animal; D045222:Electron Transport Chain Complex Proteins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D024101:Mitochondrial Proteins / Q000378:metabolism; D009154:Mutation; D010006:Osteoblasts / Q000378:metabolism* / Q000473:pathology; D010024:Osteoporosis / Q000235:genetics* / Q000378:metabolism
chemical_list: D045222:Electron Transport Chain Complex Proteins; D024101:Mitochondrial Proteins; D000074002:DNA Polymerase gamma; C498803:Polg protein, mouse