Ageing and ovarian stimulation modulate the relative levels of transcript abundance of oocyte DNA repair genes during th
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REPRODUCTIVE PHYSIOLOGY AND DISEASE
Ageing and ovarian stimulation modulate the relative levels of transcript abundance of oocyte DNA repair genes during the germinal vesicle-metaphase II transition in mice Fabrizzio Horta 1,2
&
Aravind Ravichandran 1 & Sally Catt 1 & Beverley Vollenhoven 1,2,3 & Peter Temple-Smith 1
Received: 27 July 2020 / Accepted: 13 October 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Purpose Oocyte quality and reproductive outcome are negatively affected by advanced maternal age, ovarian stimulation and method of oocyte maturation during assisted reproduction; however, the mechanisms responsible for these associations are not fully understood. The aim of this study was to compare the effects of ageing, ovarian stimulation and in-vitro maturation on the relative levels of transcript abundance of genes associated with DNA repair during the transition of germinal vesicle (GV) to metaphase II (MII) stages of oocyte development. Methods The relative levels of transcript abundance of 90 DNA repair-associated genes was compared in GV-stage and MIIstage oocytes from unstimulated and hormone-stimulated ovaries from young (5–8-week-old) and old (42–45-week-old) C57BL6 mice. Ovarian stimulation was conducted using pregnant mare serum gonadotropin (PMSG) or anti-inhibin serum (AIS). DNA damage response was quantified by immunolabeling of the phosphorylated histone variant H2AX (γH2AX). Results The relative transcript abundance in DNA repair genes was significantly lower in MII oocytes compared to GV oocytes in young unstimulated and PMSG stimulated but was higher in AIS-stimulated mice. Interestingly, an increase in the relative level of transcript abundance of DNA repair genes was observed in MII oocytes from older mice in unstimulated, PMSGstimulated and AIS-stimulated mice. Decreased γH2AX levels were found in both GV oocytes (82.9%) and MII oocytes (37.5%) during ageing in both ovarian stimulation types used (PMSG/AIS; p < 0.05). Conclusions In conclusion, DNA repair relative levels of transcript abundance are altered by maternal age and the method of ovarian stimulation during the GV-MII transition in oocytes. Keywords Oocyte DNA repair . Female ageing . mRNA . Ovarian stimulation . In vitro maturation
Introduction Prophase arrest at the germinal vesicle (GV) stage in oocytes is significant for two reasons: (I) the accumulation of mRNA Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10815-020-01981-6) contains supplementary material, which is available to authorized users. * Fabrizzio Horta [email protected] 1
Education Program in Reproduction and Development, EPRD, Department of Obstetrics and Gynecology, School of Clinical Science, Monash University, Melbourne, VIC 3168, Australia
2
Monash IVF, Melbourne, VIC 3168, Australia
3
Women’s and Newborn Program, Monash Health, Melbourne, Australia
transcripts and proteins is required for oocyte maturation and early embryo development [1–3] and (II) e
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