title: mtDNA point mutations are present at various levels of heteroplasmy in human oocytes.
abstract: Little is known about the load of mutations and polymorphisms in the mitochondrial DNA (mtDNA) of human oocytes and the possible effect these mutations may have during life. To investigate this, we optimised at the single cell level the recently developed method to screen the entire mtDNA for mainly heteroplasmic mutations by denaturing high performance liquid chromatography analysis. This method is sensitive (approximately 1% heteroplasmy detectable), specific and rapid. The entire mtDNA of 26 oocytes of 13 women was screened by this method. Ten different heteroplasmic mutations, of which only one was located in the D-loop and two were observed twice, were detected in seven oocytes with mutation loads ranging from <5% to 50%. From eight women >1 oocyte was received and in four of them heteroplasmic differences between oocytes of the same woman were observed. In one of these four, two homoplasmic D-loop variants were also detected. Additionally, four oocytes of a single woman were sequenced using the MitoChip (which lacks the D-loop region), but all sequences were identical. It is concluded that heteroplasmic mtDNA mutations are common in oocytes and that, depending on the position and mutation load, they might increase the risk of developing OXPHOS disease early or later in life.
authors.affiliation: Department of Genetics and Cell Biology, University of Maastricht, The Netherlands.
authors.forename: Lorraine
authors.initials: L
authors.lastname: Jacobs
authors.forename: Mike
authors.initials: M
authors.lastname: Gerards
authors.forename: Patrick
authors.initials: P
authors.lastname: Chinnery
authors.forename: John
authors.initials: J
authors.lastname: Dumoulin
authors.forename: Ireneaus
authors.initials: I
authors.lastname: de Coo
authors.forename: Joep
authors.initials: J
authors.lastname: Geraedts
authors.forename: Hubert
authors.initials: H
authors.lastname: Smeets
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004252:DNA Mutational Analysis / Q000379:methods*; D004272:DNA, Mitochondrial / Q000378:metabolism*; D005260:Female; D030342:Genetic Diseases, Inborn / Q000175:diagnosis / Q000235:genetics; D020022:Genetic Predisposition to Disease; D006801:Humans; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism; D028361:Mitochondrial Diseases / Q000175:diagnosis / Q000235:genetics; D009691:Nucleic Acid Denaturation; D020411:Oligonucleotide Array Sequence Analysis; D009865:Oocytes / Q000378:metabolism*; D010085:Oxidative Phosphorylation; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011296:Prenatal Diagnosis / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004272:DNA, Mitochondrial