{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T12:36:00Z","timestamp":1791894960634,"version":"4.3.4"},"reference-count":61,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2015,10,15]],"date-time":"2015-10-15T00:00:00Z","timestamp":1444867200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2015,10,15]],"date-time":"2015-10-15T00:00:00Z","timestamp":1444867200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Oxidative phosphorylation defects in human tissues are often challenging to quantify due to a mosaic pattern of deficiency. Biochemical assays are difficult to interpret due to the varying enzyme deficiency levels found in individual cells. Histochemical analysis allows semi-quantitative assessment of complex II and complex IV activities, but there is no validated histochemical assay to assess complex I activity which is frequently affected in mitochondrial pathology. To help improve the diagnosis of mitochondrial disease and to study the mechanisms underlying mitochondrial abnormalities in disease, we have developed a quadruple immunofluorescent technique enabling the quantification of key respiratory chain subunits of complexes I and IV, together with an indicator of mitochondrial mass and a cell membrane marker. This assay gives precise and objective quantification of protein abundance in large numbers of individual muscle fibres. By assessing muscle biopsies from subjects with a range of different mitochondrial genetic defects we have demonstrated that specific genotypes exhibit distinct biochemical signatures in muscle, providing evidence for the diagnostic use of the technique, as well as insight into the underlying molecular pathology. Stringent testing for reproducibility and sensitivity confirms the potential value of the technique for mechanistic studies of disease and in the evaluation of therapeutic approaches.<\/jats:p>","DOI":"10.1038\/srep15037","type":"journal-article","created":{"date-parts":[[2015,10,15]],"date-time":"2015-10-15T09:11:09Z","timestamp":1444900269000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":119,"title":["A novel immunofluorescent assay to investigate oxidative phosphorylation deficiency in mitochondrial myopathy: understanding mechanisms and improving diagnosis"],"prefix":"10.1038","volume":"5","author":[{"given":"Mariana C.","family":"Rocha","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John P.","family":"Grady","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"Gr\u00fcnewald","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amy","family":"Vincent","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip F.","family":"Dobson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert W.","family":"Taylor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doug M.","family":"Turnbull","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karolina A.","family":"Rygiel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2015,10,15]]},"reference":[{"key":"BFsrep15037_CR1","doi-asserted-by":"publisher","first-page":"753","DOI":"10.1002\/ana.24362","volume":"77","author":"GS Gorman","year":"2015","unstructured":"Gorman, G. S. et al. Prevalence of nuclear and mtDNA mutations related to adult mitochondrial disease. Ann Neurol 77, 753\u2013759 (2015).","journal-title":"Ann Neurol"},{"key":"BFsrep15037_CR2","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1002\/path.3028","volume":"226","author":"LC Greaves","year":"2012","unstructured":"Greaves, L. C., Reeve, A. K., Taylor, R. W. & Turnbull, D. M. Mitochondrial DNA and disease. J Pathol 226, 274\u2013286 (2012).","journal-title":"J Pathol"},{"key":"BFsrep15037_CR3","doi-asserted-by":"publisher","first-page":"481","DOI":"10.1002\/ana.22109","volume":"69","author":"GR Campbell","year":"2011","unstructured":"Campbell, G. R. et al. Mitochondrial DNA deletions and neurodegeneration in multiple sclerosis. Ann Neurol 69, 481\u2013492 (2011).","journal-title":"Ann Neurol"},{"key":"BFsrep15037_CR4","doi-asserted-by":"publisher","first-page":"325","DOI":"10.1093\/brain\/116.2.325","volume":"116","author":"A Oldfors","year":"1993","unstructured":"Oldfors, A., Larsson, N.-G., Lindberg, C. & Holme, E. Mitochondrial DNA deletions in inclusion body myositis. Brain 116, 325\u2013336 (1993).","journal-title":"Brain"},{"key":"BFsrep15037_CR5","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1016\/j.ajhg.2007.09.018","volume":"82","author":"AK Reeve","year":"2008","unstructured":"Reeve, A. K. et al. Nature of mitochondrial DNA deletions in substantia nigra neurons. Am J Hum Genet 82, 228\u2013235 (2008).","journal-title":"Am J Hum Genet"},{"key":"BFsrep15037_CR6","doi-asserted-by":"publisher","first-page":"576","DOI":"10.1097\/WCO.0000000000000126","volume":"27","author":"RD Pitceathly","year":"2014","unstructured":"Pitceathly, R. D. & McFarland, R. Mitochondrial myopathies in adults and children: management and therapy development. Curr Opin Neurol 27, 576\u2013582 (2014).","journal-title":"Curr Opin Neurol"},{"key":"BFsrep15037_CR7","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1016\/j.nmd.2003.12.004","volume":"14","author":"RW Taylor","year":"2004","unstructured":"Taylor, R. W., Schaefer, A. M., Barron, M. J., McFarland, R. & Turnbull, D. M. The diagnosis of mitochondrial muscle disease. Neuromuscul Disord 14, 237\u2013245 (2004).","journal-title":"Neuromuscul Disord"},{"key":"BFsrep15037_CR8","doi-asserted-by":"publisher","first-page":"829","DOI":"10.1016\/S1474-4422(10)70116-2","volume":"9","author":"R McFarland","year":"2010","unstructured":"McFarland, R., Taylor, R. W. & Turnbull, D. M. A neurological perspective on mitochondrial disease. The Lancet Neurology 9, 829\u2013840 (2010).","journal-title":"The Lancet Neurology"},{"key":"BFsrep15037_CR9","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/0022-510X(83)90125-9","volume":"60","author":"MA Johnson","year":"1983","unstructured":"Johnson, M. A., Turnbull, D. M., Dick, D. J. & Sherratt, H. S. A partial deficiency of cytochrome c oxidase in chronic progressive external ophthalmoplegia. J Neurol Sci 60, 31\u201353 (1983).","journal-title":"J Neurol Sci"},{"key":"BFsrep15037_CR10","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1093\/hmg\/3.1.13","volume":"3","author":"M Sciacco","year":"1994","unstructured":"Sciacco, M., Bonilla, E., Schon, E. A., DiMauro, S. & Moraes, C. T. Distribution of wild-type and common deletion forms of mtDNA in normal and respiration-deficient muscle fibers from patients with mitochondrial myopathy. Hum Mol Genet 3, 13\u201319 (1994).","journal-title":"Hum Mol Genet"},{"key":"BFsrep15037_CR11","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/S0091-679X(06)80004-X","volume":"80","author":"DM Kirby","year":"2007","unstructured":"Kirby, D. M., Thorburn, D. R., Turnbull, D. M. & Taylor, R. W. Biochemical assays of respiratory chain complex activity. Methods Cell Biol 80, 93\u2013119 (2007).","journal-title":"Methods Cell Biol"},{"key":"BFsrep15037_CR12","doi-asserted-by":"publisher","first-page":"545","DOI":"10.1007\/BF01753355","volume":"21","author":"SL Old","year":"1989","unstructured":"Old, S. L. & Johnson, M. A. Methods of microphotometric assay of succinate dehydrogenase and cytochrome c oxidase activities for use on human skeletal muscle. Histochem J 21, 545\u2013555 (1989).","journal-title":"Histochem J"},{"key":"BFsrep15037_CR13","doi-asserted-by":"publisher","first-page":"509","DOI":"10.1016\/S0076-6879(96)64045-2","volume":"264","author":"M Sciacco","year":"1996","unstructured":"Sciacco, M. & Bonilla, E. Cytochemistry and immunocytochemistry of mitochondria in tissue sections. Methods Enzymol 264, 509\u2013521 (1996).","journal-title":"Methods Enzymol"},{"key":"BFsrep15037_CR14","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1369\/jhc.2009.953554","volume":"58","author":"KR Choudhury","year":"2010","unstructured":"Choudhury, K. R., Yagle, K. J., Swanson, P. E., Krohn, K. A. & Rajendran, J. G. A robust automated measure of average antibody staining in immunohistochemistry images. J Histochem Cytochem 58, 95\u2013107 (2010).","journal-title":"J Histochem Cytochem"},{"key":"BFsrep15037_CR15","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1186\/1746-1596-7-42","volume":"7","author":"AE Rizzardi","year":"2012","unstructured":"Rizzardi, A. E. et al. Quantitative comparison of immunohistochemical staining measured by digital image analysis versus pathologist visual scoring. Diagn Pathol 7, 42 (2012).","journal-title":"Diagn Pathol"},{"key":"BFsrep15037_CR16","doi-asserted-by":"publisher","first-page":"499","DOI":"10.1007\/s10545-006-0362-4","volume":"29","author":"RJ Janssen","year":"2006","unstructured":"Janssen, R. J., Nijtmans, L. G., van den Heuvel, L. P. & Smeitink, J. A. Mitochondrial complex I: structure, function and pathology. J Inherit Metab Dis 29, 499\u2013515 (2006).","journal-title":"J Inherit Metab Dis"},{"key":"BFsrep15037_CR17","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1002\/(SICI)1098-1004(200002)15:2<123::AID-HUMU1>3.0.CO;2-P","volume":"15","author":"JL Loeffen","year":"2000","unstructured":"Loeffen, J. L. et al. Isolated complex I deficiency in children: clinical, biochemical and genetic aspects. Hum Mutat 15, 123\u2013134 (2000).","journal-title":"Hum Mutat"},{"key":"BFsrep15037_CR18","doi-asserted-by":"publisher","first-page":"288","DOI":"10.1111\/nan.12149","volume":"41","author":"KA Rygiel","year":"2015","unstructured":"Rygiel, K. A. et al. Mitochondrial and inflammatory changes in sporadic Inclusion Body Myositis. Neuropathol Appl Neurobiol 41, 288\u2013303 (2015).","journal-title":"Neuropathol Appl Neurobiol"},{"key":"BFsrep15037_CR19","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1016\/S0022-510X(03)00039-X","volume":"211","author":"PF Chinnery","year":"2003","unstructured":"Chinnery, P. F., Howel, D., Turnbull, D. M. & Johnson, M. A. Clinical progression of mitochondrial myopathy is associated with the random accumulation of cytochrome c oxidase negative skeletal muscle fibres. J Neurol Sci 211, 63\u201366 (2003).","journal-title":"J Neurol Sci"},{"key":"BFsrep15037_CR20","first-page":"e75","volume":"41","author":"R Horvath","year":"2004","unstructured":"Horvath, R. et al. Spontaneous recovery of a childhood onset mitochondrial myopathy caused by a stop mutation in the mitochondrial cytochrome c oxidase III gene. Lancet Neurol 41, e75 (2004).","journal-title":"Lancet Neurol"},{"key":"BFsrep15037_CR21","doi-asserted-by":"publisher","first-page":"3391","DOI":"10.1093\/brain\/awl282","volume":"129","author":"T Taivassalo","year":"2006","unstructured":"Taivassalo, T. et al. Endurance training and detraining in mitochondrial myopathies due to single large-scale mtDNA deletions. Brain 129, 3391\u20133401 (2006).","journal-title":"Brain"},{"key":"BFsrep15037_CR22","doi-asserted-by":"publisher","first-page":"2832","DOI":"10.1093\/brain\/awn252","volume":"131","author":"JL Murphy","year":"2008","unstructured":"Murphy, J. L. et al. Resistance training in patients with single, large-scale deletions of mitochondrial DNA. Brain 131, 2832\u20132840 (2008).","journal-title":"Brain"},{"key":"BFsrep15037_CR23","doi-asserted-by":"publisher","first-page":"3038","DOI":"10.1128\/MCB.00025-10","volume":"30","author":"E Perales-Clemente","year":"2010","unstructured":"Perales-Clemente, E. et al. Five entry points of the mitochondrially encoded subunits in mammalian complex I assembly. Mol Cell Biol 30, 3038\u20133047 (2010).","journal-title":"Mol Cell Biol"},{"key":"BFsrep15037_CR24","unstructured":"R: A language and environment for statistical computing (R Foundation for Statistical Computing, Vienna, Austria, 2014)."},{"key":"BFsrep15037_CR25","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1111\/j.2517-6161.1964.tb00553.x","volume":"26","author":"GEP Box","year":"1964","unstructured":"Box, G. E. P. & Cox, D. R. An Analysis of Transformations. J R Stat Soc Series B Stat Methodol 26, 211\u2013252 (1964).","journal-title":"J R Stat Soc Series B Stat Methodol"},{"key":"BFsrep15037_CR26","doi-asserted-by":"publisher","first-page":"591","DOI":"10.1093\/biomet\/52.3-4.591","volume":"52","author":"SS Shapiro","year":"1965","unstructured":"Shapiro, S. S. & Wilk, M. B. An Analysis of Variance Test for Normality (Complete Samples). Biometrika 52, 591\u2212611 (1965).","journal-title":"Biometrika"},{"key":"BFsrep15037_CR27","unstructured":"Goodness-of-fit techniques. (Marcel Dekker, Inc., 1986)."},{"key":"BFsrep15037_CR28","doi-asserted-by":"publisher","first-page":"690","DOI":"10.1016\/j.nmd.2012.04.003","volume":"22","author":"JL Murphy","year":"2012","unstructured":"Murphy, J. L. et al. Cytochrome c oxidase-intermediate fibres: importance in understanding the pathogenesis and treatment of mitochondrial myopathy. Neuromuscul Disord 22, 690\u2013698 (2012).","journal-title":"Neuromuscul Disord"},{"key":"BFsrep15037_CR29","doi-asserted-by":"publisher","first-page":"148","DOI":"10.1152\/japplphysiol.01022.2012","volume":"114","author":"J Mula","year":"2013","unstructured":"Mula, J., Lee, J. D., Liu, F., Yang, L. & Peterson, C. A. Automated image analysis of skeletal muscle fiber cross-sectional area. J Appl Physiol (1985) 114, 148\u2013155 (2013).","journal-title":"J Appl Physiol (1985)"},{"key":"BFsrep15037_CR30","doi-asserted-by":"publisher","first-page":"1079","DOI":"10.1038\/modpathol.3880440","volume":"14","author":"TA Thomson","year":"2001","unstructured":"Thomson, T. A. et al. HER-2\/neu in breast cancer: interobserver variability and performance of immunohistochemistry with 4 antibodies compared with fluorescent in situ hybridization. Mod Pathol 14, 1079\u20131086 (2001).","journal-title":"Mod Pathol"},{"key":"BFsrep15037_CR31","doi-asserted-by":"publisher","first-page":"272","DOI":"10.1369\/0022155413477661","volume":"61","author":"AW Alvarenga","year":"2013","unstructured":"Alvarenga, A. W. et al. A comparison between manual and automated evaluations of tissue microarray patterns of protein expression. J Histochem Cytochem 61, 272\u2013282 (2013).","journal-title":"J Histochem Cytochem"},{"key":"BFsrep15037_CR32","doi-asserted-by":"publisher","first-page":"2969","DOI":"10.1093\/brain\/awn239","volume":"131","author":"S Kloppel","year":"2008","unstructured":"Kloppel, S. et al. Accuracy of dementia diagnosis: a direct comparison between radiologists and a computerized method. Brain 131, 2969\u20132974 (2008).","journal-title":"Brain"},{"key":"BFsrep15037_CR33","doi-asserted-by":"publisher","first-page":"172","DOI":"10.1136\/jcp.2008.061267","volume":"62","author":"B De Paepe","year":"2009","unstructured":"De Paepe, B. et al. Immunohistochemical analysis of the oxidative phosphorylation complexes in skeletal muscle from patients with mitochondrial DNA encoded tRNA gene defects. J Clin Pathol 62, 172\u2013176 (2009).","journal-title":"J Clin Pathol"},{"key":"BFsrep15037_CR34","doi-asserted-by":"publisher","first-page":"1281","DOI":"10.1177\/002215540205001001","volume":"50","author":"BJ Hanson","year":"2002","unstructured":"Hanson, B. J., Capaldi, R. A., Marusich, M. F. & Sherwood, S. W. An immunocytochemical approach to detection of mitochondrial disorders. J Histochem Cytochem 50, 1281\u20131288 (2002).","journal-title":"J Histochem Cytochem"},{"key":"BFsrep15037_CR35","doi-asserted-by":"publisher","first-page":"310","DOI":"10.1016\/j.jneumeth.2009.08.020","volume":"184","author":"DJ Mahad","year":"2009","unstructured":"Mahad, D. J. et al. Detection of cytochrome c oxidase activity and mitochondrial proteins in single cells. J Neurosci Methods 184, 310\u2013319 (2009).","journal-title":"J Neurosci Methods"},{"key":"BFsrep15037_CR36","doi-asserted-by":"publisher","first-page":"591","DOI":"10.1093\/brain\/123.3.591","volume":"123","author":"S Rahman","year":"2000","unstructured":"Rahman, S. et al. Cytochrome oxidase immunohistochemistry: clues for genetic mechanisms. Brain 123 Pt 3, 591\u2013600 (2000).","journal-title":"Brain"},{"key":"BFsrep15037_CR37","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1212\/WNL.41.2_Part_1.300","volume":"41","author":"HJ Tritschler","year":"1991","unstructured":"Tritschler, H. J. et al. Differential diagnosis of fatal and benign cytochrome c oxidase-deficient myopathies of infancy: an immunohistochemical approach. Neurology 41, 300\u2013305 (1991).","journal-title":"Neurology"},{"key":"BFsrep15037_CR38","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1016\/j.jneumeth.2014.05.026","volume":"232","author":"A Grunewald","year":"2014","unstructured":"Grunewald, A. et al. Quantitative quadruple-label immunofluorescence of mitochondrial and cytoplasmic proteins in single neurons from human midbrain tissue. J Neurosci Methods 232, 143\u2013149 (2014).","journal-title":"J Neurosci Methods"},{"key":"BFsrep15037_CR39","doi-asserted-by":"publisher","first-page":"858","DOI":"10.1016\/S0960-8966(02)00047-0","volume":"12","author":"JL Elson","year":"2002","unstructured":"Elson, J. L., Samuels, D. C., Johnson, M. A., Turnbull, D. M. & Chinnery, P. F. The length of cytochrome c oxidase-negative segments in muscle fibres in patients with mtDNA myopathy. Neuromuscul Disord 12, 858\u2013864 (2002).","journal-title":"Neuromuscul Disord"},{"key":"BFsrep15037_CR40","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1002\/mus.880150213","volume":"15","author":"T Matsuoka","year":"1992","unstructured":"Matsuoka, T., Goto, Y., Hasegawa, H. & Nonaka, I. Segmental cytochrome c-oxidase deficiency in CPEO: teased muscle fiber analysis. Muscle Nerve 15, 209\u2013213 (1992).","journal-title":"Muscle Nerve"},{"key":"BFsrep15037_CR41","doi-asserted-by":"publisher","first-page":"4612","DOI":"10.1093\/hmg\/ddu176","volume":"23","author":"G Campbell","year":"2014","unstructured":"Campbell, G., Krishnan, K. J., Deschauer, M., Taylor, R. W. & Turnbull, D. M. Dissecting the mechanisms underlying the accumulation of mitochondrial DNA deletions in human skeletal muscle. Hum Mol Genet 23, 4612\u20134620 (2014).","journal-title":"Hum Mol Genet"},{"key":"BFsrep15037_CR42","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1016\/j.bbamcr.2008.04.015","volume":"1793","author":"M Lazarou","year":"2009","unstructured":"Lazarou, M., Thorburn, D. R., Ryan, M. T. & McKenzie, M. Assembly of mitochondrial complex I and defects in disease. Biochim Biophys Acta 1793, 78\u201388 (2009).","journal-title":"Biochim Biophys Acta"},{"key":"BFsrep15037_CR43","doi-asserted-by":"publisher","first-page":"200","DOI":"10.1016\/j.bbamcr.2008.05.028","volume":"1793","author":"E Fern\u00e1ndez-Vizarra","year":"2009","unstructured":"Fern\u00e1ndez-Vizarra, E., Tiranti, V. & Zeviani, M. Assembly of the oxidative phosphorylation system in humans: What we have learned by studying its defects. Biochim Biophys Acta 1793, 200\u2013211 (2009).","journal-title":"Biochim Biophys Acta"},{"key":"BFsrep15037_CR44","doi-asserted-by":"publisher","first-page":"331","DOI":"10.1042\/BJ20040469","volume":"382","author":"F Xu","year":"2004","unstructured":"Xu, F., Morin, C., Mitchell, G., Ackerley, C. & Robinson, B. H. The role of the LRPPRC (leucine-rich pentatricopeptide repeat cassette) gene in cytochrome oxidase assembly: mutation causes lowered levels of COX (cytochrome c oxidase) I and COX III mRNA. Biochem J 382, 331\u2013336 (2004).","journal-title":"Biochem J"},{"key":"BFsrep15037_CR45","first-page":"481","volume":"53","author":"F Merante","year":"1993","unstructured":"Merante, F. et al. A biochemically distinct form of cytochrome oxidase (COX) deficiency in the Saguenay-Lac-Saint-Jean region of Quebec. Am J Hum Genet 53, 481\u2013487 (1993).","journal-title":"Am J Hum Genet"},{"key":"BFsrep15037_CR46","doi-asserted-by":"publisher","first-page":"605","DOI":"10.1073\/pnas.242716699","volume":"100","author":"VK Mootha","year":"2003","unstructured":"Mootha, V. K. et al. Identification of a gene causing human cytochrome c oxidase deficiency by integrative genomics. Proc Natl Acad Sci USA 100, 605\u2013610 (2003).","journal-title":"Proc Natl Acad Sci USA"},{"key":"BFsrep15037_CR47","doi-asserted-by":"publisher","first-page":"769","DOI":"10.1038\/ejhg.2011.18","volume":"19","author":"H Swalwell","year":"2011","unstructured":"Swalwell, H. et al. Respiratory chain complex I deficiency caused by mitochondrial DNA mutations. Eur J Hum Genet 19, 769\u2013775 (2011).","journal-title":"Eur J Hum Genet"},{"key":"BFsrep15037_CR48","doi-asserted-by":"publisher","first-page":"784","DOI":"10.1136\/jmg.2004.020537","volume":"41","author":"DM Kirby","year":"2004","unstructured":"Kirby, D. M. et al. Mutations of the mitochondrial ND1 gene as a cause of MELAS. J Med Genet 41, 784\u2013789 (2004).","journal-title":"J Med Genet"},{"key":"BFsrep15037_CR49","first-page":"1147","volume":"48","author":"K Huoponen","year":"1991","unstructured":"Huoponen, K., Vilkki, J., Aula, P., Nikoskelainen, E. K. & Savontaus, M. L. A new mtDNA mutation associated with Leber hereditary optic neuroretinopathy. Am J Hum Genet 48, 1147\u20131153 (1991).","journal-title":"Am J Hum Genet"},{"key":"BFsrep15037_CR50","doi-asserted-by":"publisher","first-page":"895","DOI":"10.1042\/CS20140705","volume":"128","author":"GS Gorman","year":"2015","unstructured":"Gorman, G. S. et al. Novel MTND1 mutations cause isolated exercise intolerance, complex I deficiency and increased assembly factor expression. Clin Sci (Lond) 128, 895\u2013904 (2015).","journal-title":"Clin Sci (Lond)"},{"key":"BFsrep15037_CR51","doi-asserted-by":"publisher","first-page":"323","DOI":"10.1093\/brain\/awt321","volume":"137","author":"JP Grady","year":"2014","unstructured":"Grady, J. P. et al. Disease progression in patients with single, large-scale mitochondrial DNA deletions. Brain 137, 323\u2013334 (2014).","journal-title":"Brain"},{"key":"BFsrep15037_CR52","doi-asserted-by":"publisher","first-page":"e68","DOI":"10.1093\/nar\/gnf067","volume":"30","author":"L He","year":"2002","unstructured":"He, L. et al. Detection and quantification of mitochondrial DNA deletions in individual cells by real-time PCR. Nucleic Acids Res 30, e68 (2002).","journal-title":"Nucleic Acids Res"},{"key":"BFsrep15037_CR53","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1016\/0022-510X(92)90025-G","volume":"110","author":"A Oldfors","year":"1992","unstructured":"Oldfors, A. et al. Mitochondrial DNA deletions and cytochrome c oxidase deficiency in muscle fibres. J Neurol Sci 110, 169\u2013177 (1992).","journal-title":"J Neurol Sci"},{"key":"BFsrep15037_CR54","doi-asserted-by":"publisher","first-page":"3392","DOI":"10.1093\/brain\/aws231","volume":"135","author":"RD Pitceathly","year":"2012","unstructured":"Pitceathly, R. D. et al. Adults with RRM2B-related mitochondrial disease have distinct clinical and molecular characteristics. Brain 135, 3392\u20133403 (2012).","journal-title":"Brain"},{"key":"BFsrep15037_CR55","doi-asserted-by":"publisher","first-page":"9906","DOI":"10.1038\/srep09906","volume":"5","author":"KA Rygiel","year":"2015","unstructured":"Rygiel, K. A., Grady, J. P., Taylor, R. W., Tuppen, H. A. L. & Turnbull, D. M. Triplex real-time PCR\u2013an improved method to detect a wide spectrum of mitochondrial DNA deletions in single cells. Sci Rep 5, 9906, 9910.1038\/srep09906 (2015).","journal-title":"Sci Rep"},{"key":"BFsrep15037_CR56","doi-asserted-by":"publisher","first-page":"401","DOI":"10.1042\/bj3180401","volume":"318","author":"AM James","year":"1996","unstructured":"James, A. M., Wei, Y. H., Pang, C. Y. & Murphy, M. P. Altered mitochondrial function in fibroblasts containing MELAS or MERRF mitochondrial DNA mutations. Biochem J 318 (Pt 2), 401\u2013407 (1996).","journal-title":"Biochem J"},{"key":"BFsrep15037_CR57","doi-asserted-by":"publisher","first-page":"936","DOI":"10.1136\/jnnp-2012-303528","volume":"84","author":"V Nesbitt","year":"2013","unstructured":"Nesbitt, V. et al. The UK MRC Mitochondrial Disease Patient Cohort Study: clinical phenotypes associated with the m.3243A&gt;G mutation--implications for diagnosis and management. J Neurol Neurosurg Psychiatry 84, 936\u2013938 (2013).","journal-title":"J Neurol Neurosurg Psychiatry"},{"key":"BFsrep15037_CR58","doi-asserted-by":"publisher","first-page":"637","DOI":"10.1006\/bbrc.1997.6496","volume":"233","author":"A Shaag","year":"1997","unstructured":"Shaag, A., Saada, A., Steinberg, A., Navon, P. & Elpeleg, O. N. Mitochondrial encephalomyopathy associated with a novel mutation in the mitochondrial tRNA(leu)(UUR) gene (A3243T). Biochem Biophys Res Commun 233, 637\u2013639 (1997).","journal-title":"Biochem Biophys Res Commun"},{"key":"BFsrep15037_CR59","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1007\/s00403-005-0578-4","volume":"297","author":"AY Goedkoop","year":"2005","unstructured":"Goedkoop, A. Y. et al. Digital image analysis for the evaluation of the inflammatory infiltrate in psoriasis. Arch Dermatol Res 297, 51\u201359 (2005).","journal-title":"Arch Dermatol Res"},{"key":"BFsrep15037_CR60","doi-asserted-by":"publisher","first-page":"575","DOI":"10.1177\/002215540305100503","volume":"51","author":"EM Brey","year":"2003","unstructured":"Brey, E. M. et al. Automated selection of DAB-labeled tissue for immunohistochemical quantification. J Histochem Cytochem 51, 575\u2013584 (2003).","journal-title":"J Histochem Cytochem"},{"key":"BFsrep15037_CR61","doi-asserted-by":"publisher","first-page":"741","DOI":"10.1136\/adc.56.10.741","volume":"56","author":"J Egger","year":"1981","unstructured":"Egger, J., Lake, B. D. & Wilson, J. Mitochondrial cytopathy. A multisystem disorder with ragged red fibres on muscle biopsy. Arch Dis Child 56, 741\u2013752 (1981).","journal-title":"Arch Dis Child"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep15037.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep15037","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep15037.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,6,11]],"date-time":"2024-06-11T16:38:38Z","timestamp":1718123918000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep15037"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,10,15]]},"references-count":61,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2015,12,7]]}},"alternative-id":["BFsrep15037"],"URL":"https:\/\/doi.org\/10.1038\/srep15037","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,10,15]]},"assertion":[{"value":"14 July 2015","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 September 2015","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"15 October 2015","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"15037"}}