Intra-individual purifying selection on mitochondrial DNA variants during human oogenesis.
Does selection for mtDNA mutations occur in human oocytes?



We provide statistical evidence in favor of the existence of purifying selection for mtDNA mutations in human oocytes acting between the expulsion of the first and second polar bodies (PBs).



Several lines of evidence in Metazoa, including humans, indicate that variation within the germline of mitochondrial genomes is under purifying selection. The presence of this internal selection filter in the germline has important consequences for the evolutionary trajectory of mtDNA. However, the nature and localization of this internal filter are still unclear while several hypotheses are proposed in the literature.



In this study, 60 mitochondrial genomes were sequenced from 17 sets of oocytes, first and second PBs, and peripheral blood taken from nine women between 38 and 43 years of age.



Whole genome amplification was performed only on the single cell samples and Sanger sequencing was performed on amplicons. The comparison of variant profiles between first and second PB sequences showed no difference in substitution rates but displayed instead a sharp difference in pathogenicity scores of protein-coding sequences using three different metrics (MutPred, Polyphen and SNPs&GO).



Unlike the first, second PBs showed no significant differences in pathogenic scores with blood and oocyte sequences. This suggests that a filtering mechanism for disadvantageous variants operates during oocyte development between the expulsion of the first and second PB.



N/A.



The sample size is small and further studies are needed before this approach can be used in clinical practice. Studies on a model organism would allow the sample size to be increased.



This work opens the way to the study of the correlation between mtDNA mutations, mitochondrial capacity and viability of oocytes.



This work was supported by a SISMER grant. Laboratory facilities and skills were freely provided by SISMER, and by the Alma Mater Studiorum, University of Bologna. The authors have no conflict of interest to disclose.
Department of Biological, Geological and Environmental Sciences, University of Bologna, Bologna 40126, Italy.
Sara
S
De Fanti
Institute of Atmospheric Pollution Research, National Research Council, C/O Physics Department, University of Bari 'Aldo Moro', Bari 70132, Italy.
Saverio
S
Vicario
Medical Genetics Unit, S. Orsola Hospital, University of Bologna, Bologna 40126, Italy.|Current address: Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, USA.
Martin
M
Lang
Department of Bioscience, Biotechnology and Biopharmaceutics, University of Bari 'Aldo Moro',Bari70132, Italy.
Domenico
D
Simone
Reproductive Medicine Unit, S.I.S.Me.R., Bologna 40138, Italy.
Cristina
C
Magli
Department of Biological, Geological and Environmental Sciences, University of Bologna, Bologna 40126, Italy.
Donata
D
Luiselli
Reproductive Medicine Unit, S.I.S.Me.R., Bologna 40138, Italy.
Luca
L
Gianaroli
Medical Genetics Unit, S. Orsola Hospital, University of Bologna, Bologna 40126, Italy.
Giovanni
G
Romeo
D000328:Adult; D004272:DNA, Mitochondrial / Q000235:genetics*; D005260:Female; D054629:Genome, Mitochondrial; D006801:Humans; D008928:Mitochondria / Q000235:genetics*; D009154:Mutation; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D009866:Oogenesis / Q000235:genetics*
D004272:DNA, Mitochondrial
first and second polar body; heteroplasmy; human oocytes; mtDNA variants; purifying selection