In vitro compaction of germinal vesicle chromatin is beneficial to survival of vitrified cat oocytes.
The immature cat oocyte contains a large-sized germinal vesicle (GV) with decondensed chromatin that is highly susceptible to cryo-damage. The aim of the study was to explore an alternative to conventional cryopreservation by examining the influence of GV chromatin compaction using resveratrol (Res) exposure (a histone deacetylase enhancer) on oocyte survival during vitrification. In Experiment 1, denuded oocytes were exposed to 0, 0.5, 1.0 or 1.5 mmol/l Res for 1.5 h and then evaluated for chromatin structure or cultured to assess oocyte meiotic and developmental competence in vitro. Exposure to 1.0 or 1.5 mmol/l Res induced complete GV chromatin deacetylation and the most significant compaction. Compared to other treatments, the 1.5 mmol/l Res concentration compromised the oocyte ability to achieve metaphase II (MII) or to form a blastocyst. In Experiment 2, denuded oocytes were exposed to Res as in Experiment 1 and cultured in vitro either directly (fresh) or after vitrification. Both oocyte types then were assessed for meiotic competence, fertilizability and ability to form embryos. Vitrification exerted an overall negative influence on oocyte meiotic and developmental competence. However, ability to reach MII, achieve early first cleavage, and develop to an advanced embryo stage (8-16 cells) was improved in vitrified oocytes previously exposed to 1.0 mmol/l Res compared to all counterpart treatments. In summary, results reveal that transient epigenetic modifications associated with GV chromatin compaction induced by Res is fully reversible and beneficial to oocyte survival during vitrification. This approach has allowed the production of the first cat embryos from vitrified immature oocytes.
Comizzoli
P
P
Department of Reproductive Sciences, Center for Species Survival, Smithsonian's National Zoological Park, Washington, DC, USA. comizzolip@si.edu
Wildt
D E
DE
Pukazhenthi
B S
BS
D000818:Animals; D002415:Cats / Q000196:embryology*; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism*; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology; D046149:Embryo Culture Techniques; D005260:Female; D005307:Fertilization in Vitro; D009865:Oocytes / Q000166:cytology* / Q000502:physiology; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology
D002843:Chromatin; D003451:Cryoprotective Agents; D013267:Stilbenes; D000077185:Resveratrol