Pascal Reynier, V Brochard, Jean-François Hamel, Stéphanie Chupin, V Duranthon, Pascale May-Panloup, Valérie Desquiret-Dumas, Physiopathologie Cardiovasculaire et Mitochondriale (MITOVASC), Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Universitaire d'Angers (CHU Angers), and PRES Université Nantes Angers Le Mans (UNAM)
STUDY QUESTIONDoes ageing affect the kinetics of the mitochondrial pool during oogenesis and early embryogenesis?SUMMARY ANSWERWhile we found no age-related change during oogenesis, the kinetics of mitochondrial DNA content and the expression of the factors involved in mitochondrial biogenesis appeared to be significantly altered during embryogenesis.WHAT IS KNOWN ALREADYOocyte mitochondria are necessary for embryonic development. The morphological and functional alterations of mitochondria, as well as the qualitative and quantitative mtDNA anomalies, observed during ovarian ageing may be responsible for the alteration of oocyte competence and embryonic development.STUDY DESIGN, SIZE, DURATIONThe study, conducted from November 2016 to November 2017, used 40 mice aged 5–8 weeks and 45 mice aged 9–11 months (C57Bl6/CBA F(1)). A total of 488 immature oocytes, with a diameter ranging from 20 μm to more than 80 μm, were collected from ovaries, and 1088 mature oocytes or embryos at different developmental stages (two PN, one-cell, i.e. syngamy, two-cell, four-cell, eight-cell, morula and blastocyst) were obtained after ovarian stimulation and, for embryos, mating.PARTICIPANTS/MATERIALS, SETTING, METHODSMitochondrial DNA was quantified by quantitative PCR. We used quantitative reverse transcriptase PCR (RT-PCR) (microfluidic method) to study the relative expression of three genes involved in the key steps of embryogenesis, i.e. embryonic genome activation (HSPA1) and differentiation (CDX2 and NANOG), two mtDNA genes (CYB and ND2) and five genes essential for mitochondrial biogenesis (PPARGC1A, NRF1, POLG, TFAM and PRKAA). The statistical analysis was based on mixed linear regression models applying a logistic link function (STATA v13.1 software), with values of P MAIN RESULTS AND THE ROLE OF CHANCEDuring oogenesis, there was a significant increase in oocyte mtDNA content (P LARGE SCALE DATAN/A.LIMITATIONS, REASONS FOR CAUTIONBecause of the ethical impossibility of working on a human, this study was conducted only on a murine model. As superovulation was used, we cannot totally exclude that the differences observed were, at least partially, influenced by differences in ovarian response between young and old mice.WIDER IMPLICATIONS OF THE FINDINGSOur findings suggest a pathophysiological explanation for the link observed between mitochondria and the deterioration of oocyte quality and early embryonic development with age.STUDY FUNDING/COMPETING INTEREST(S)This work was supported by the University of Angers, France, by the French national research centres INSERM and the CNRS and, in part, by PHASE Division, INRA. There are no competing interests.