title: Flow cytometry analysis: a quantitative method for collagen VI deficiency screening.
abstract: Mutations in COL6A1, COL6A2 and COL6A3 genes result in collagen VI myopathies: Ullrich congenital muscular dystrophy (UCMD), Bethlem myopathy (BM) and intermediate phenotypes. At present, none of the existing diagnostic techniques for evaluating collagen VI expression is quantitative, and the detection of subtle changes in collagen VI expression remains challenging. We investigated flow cytometry analysis as a means of quantitatively measuring collagen VI in primary fibroblasts and compared this method with the standard method of fibroblast collagen VI immunohistochemical analysis. Eight UCMD and five BM molecularly confirmed patients were studied and compared to five controls. Flow cytometry analysis consistently detected a reduction of collagen VI of at least 60% in all UCMD cases. In BM cases the levels of collagen VI were variable but on average 20% less than controls. Flow cytometry analysis provides an alternative method for screening for collagen VI deficiency at the protein level in a quantitative, time and cost-effective manner.
authors.affiliation: Dubowitz Neuromuscular Centre, University College London Institute of Child Health, London, UK.
authors.forename: J
authors.initials: J
authors.lastname: Kim
authors.forename: C
authors.initials: C
authors.lastname: Jimenez-Mallebrera
authors.forename: A R
authors.initials: AR
authors.lastname: Foley
authors.forename: M
authors.initials: M
authors.lastname: Fernandez-Fuente
authors.forename: S C
authors.initials: SC
authors.lastname: Brown
authors.forename: S
authors.initials: S
authors.lastname: Torelli
authors.forename: L
authors.initials: L
authors.lastname: Feng
authors.forename: C A
authors.initials: CA
authors.lastname: Sewry
authors.forename: F
authors.initials: F
authors.lastname: Muntoni
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D024142:Collagen Type VI / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D008875:Middle Aged; D009136:Muscular Dystrophies / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D055815:Young Adult
chemical_list: D024142:Collagen Type VI
references.citation: J Neuropathol Exp Neurol. 2008 Feb;67(2):144-54
references.pmid: 18219255
references.citation: Cell Motil Cytoskeleton. 2007 Dec;64(12):914-25
references.pmid: 17922482
references.citation: Neuromuscul Disord. 2006 Oct;16(9-10):571-82
references.pmid: 16935502
references.citation: Neurology. 2005 Jun 14;64(11):1931-7
references.pmid: 15955946
references.citation: Clin Diagn Lab Immunol. 1994 May;1(3):299-303
references.pmid: 7496966
references.citation: Neurology. 2009 Dec 1;73(22):1883-91
references.pmid: 19949035
references.citation: Brain Dev. 1979;1(1):61-7
references.pmid: 551739
references.citation: Ann Neurol. 2010 Feb;67(2):201-8
references.pmid: 20225280
references.citation: J Biol Chem. 2008 Apr 18;283(16):10658-70
references.pmid: 18276594
references.citation: Neuromuscul Disord. 2010 Jan;20(1):57-60
references.pmid: 19854055
references.citation: Hum Mol Genet. 2005 Jan 15;14(2):279-93
references.pmid: 15563506
references.citation: Neurology. 2004 Feb 24;62(4):620-3
references.pmid: 14981181
references.citation: J Med Genet. 2005 Feb;42(2):108-20
references.pmid: 15689448
references.citation: J Biol Chem. 2008 Jul 18;283(29):20170-80
references.pmid: 18400749
references.citation: J Biol Chem. 2003 Apr 11;278(15):12601-4
references.pmid: 12556454
references.citation: Science. 2005 Sep 23;309(5743):2064-7
references.pmid: 16141372
references.citation: Neuromuscul Disord. 2005 Apr;15(4):303-10
references.pmid: 15792870
references.citation: Neuromuscul Disord. 2009 Dec;19(12):813-7
references.pmid: 19884007
references.citation: Neurology. 2008 Oct 14;71(16):1245-53
references.pmid: 18852439
references.citation: Clin Chem. 2002 Oct;48(10):1819-27
references.pmid: 12324512
references.citation: Muscle Nerve. 2002 Apr;25(4):513-9
references.pmid: 11932968
references.citation: Eur J Paediatr Neurol. 2002;6(6):309-14
references.pmid: 12401455
references.citation: Neurology. 2002 May 14;58(9):1354-9
references.pmid: 12011280
references.citation: Am J Hum Genet. 2003 Aug;73(2):355-69
references.pmid: 12840783
references.citation: Neuropediatrics. 2004 Apr;35(2):103-12
references.pmid: 15127309
references.citation: Neurology. 2008 Apr 1;70(14):1192-9
references.pmid: 18378883
references.citation: Neuromuscul Disord. 2004 Oct;14(10):635-49
references.pmid: 15351421