Oocytes recovered after ovarian tissue slow freezing have impaired H2AX phosphorylation and functional competence
Sam Sudhakaran A , Shubhashree Uppangala A , Sujith Raj Salian A , Sachin D. Honguntikar A , Ramya Nair A , Guruprasad Kalthur A and Satish Kumar Adiga A B
+ Author Affiliations
- Author Affiliations
A Division of Clinical Embryology, Department of Obstetrics and Gynaecology, Kasturba Medical College, Manipal University, Manipal-576 104, India.
B Corresponding author. Email: satish.adiga@manipal.edu
Reproduction, Fertility and Development 27(8) 1242-1248 https://doi.org/10.1071/RD14048
Submitted: 8 February 2014 Accepted: 8 May 2014 Published: 15 July 2014
Abstract
It has been shown that oocytes isolated from ovarian tissue cryopreservation acquire DNA damage during the process of freeze–thawing. Using a mouse model, here we have investigated the functional competence and phosphorylation of H2AX (γ-H2AX) in germinal vesicle (GV) and parthenogenetically activated oocytes derived from conventional ovarian tissue slow freezing and vitrification techniques. The number of GV-stage oocytes with γ-H2AX foci was not significantly different between the slow-freezing and vitrification groups. Although the in vitro maturation (IVM) potential of GV oocytes in the slow-freezing group showed a significant delay (P < 0.0001) in the process of germinal vesicle breakdown, no difference in the maturation rate was observed between the two protocols. Nevertheless, parthenogenetic activation of IVM oocytes using strontium chloride showed a significantly lower activation rate in the slow-freezing group compared with the vitrification (P < 0.05) and control (P < 0.01) groups. Importantly, H2AX phosphorylation was significantly perturbed in the slow-freezing group in comparison to the control (P < 0.05). Therefore, we conclude that impaired sensing of DNA strand breaks and repair processes are associated with the reduced functional competence of the oocytes recovered from the slow-freezing group, which may have a significant impact on the reproductive outcome.
Additional keywords: γ-H2AX, in vitro maturation, vitrification.
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