Search

Despite significant progress in cryopreservation of mammalian oocytes and embryos, many of the molecular and biochemical events that underlie this technology are poorly understood. In recent years, researchers have focused on obtaining viable oocytes that are developmentally competent. Even under the most favourable conditions, experimental approaches have achieved only limited success compared with fresh oocytes used in routine in vitro embryo production. Chilling injuries and toxic effects of the cryoprotectants are the major adverse consequences following cryoprocedures. To overcome these problems, different strategies have been developed for improving cryopreservation results. These strategies include reducing container volumes, increasing the thermal gradient, changing the cell surface/volume ratio, enhancing cryotolerance by supplementation with various additives or modifying the lipid composition of the oocyte membrane. In order to develop new strategies for reducing the various forms of stress associated with oocyte cryopreservation, it is fundamental to gain a better understanding of the major changes responsible for poor post-thaw survival. With this knowledge, we hope that oocyte cryostorage will become a fully reliable reproductive technique in the near future. [ABSTRACT FROM AUTHOR]

This study compares the developmental capacity and cryotolerance of embryos produced from oocytes of stimulated prepubertal and adult Sarda goats. Twelve prepubertal and 13 adult goats were each given 110 and 175 IU FSH, respectively, and cumulus–oocyte complexes (COCs) were collected by laparoscopic oocyte-pick-up (LOPU). After in vitromaturation, fertilisation and culture (IVMFC), blastocysts were vitrified, warmed and blastocoel re-expansion and gene expression were evaluated. Prepubertal goats produced a higher COCs number than adults (mean ± s.e.m., 89.67 ± 5.74 and 26.69 ± 3.66, respectively; P< 0.01). Lower developmental competence was demonstrated in the prepubertal oocytes as shown by a higher number of COCs discarded before IVM (21.1% and 14.7% for prepubertals and adults, respectively; P< 0.01) and IVF (23.4% v.9.1%; P< 0.01) and by the lower cleavage (55.6% and 70.3%, respectively; P< 0.01) and blastocyst rates (24.2% and 33.9%, respectively; P< 0.05). Compared with the adult, prepubertal vitrified/warmed blastocysts showed significantly (P< 0.05) lower in vitroviability, as determined by the re-expansion rate (62.5% and 40.3%). No differences were observed in the time required for blastocoel re-expansion or in cyclin B1, E-cadherin, Na/K ATPase, HSP90β and aquaporin 3 messenger RNA quantity. These results show that in vitro-produced embryos produced from prepubertal goat oocytes have a lower developmental rate and cryotolerance compared with their adult counterparts. However, we can assume that the quality of re-expanded embryos does not differ between the two groups. [ABSTRACT FROM AUTHOR]

The expression patterns of four maternal effect genes (MEG), namely zygote arrest 1 (ZAR1), maternal antigen that embryo requires (MATER), growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), were determined in ovine oocytes and in vitro-produced preimplantation embryos. The existence of ZAR1and MATERin ovine species has not been reported previously. Reverse transcription–polymerase chain reaction was performed on germinal vesicle and IVM MII oocytes, as well as in in vitrofertilised and cultured two-, four-, eight- and 12/16-cell embryos, morulae and blastocysts. Quantification of gene expression by real-time polymerase chain reaction showed the highest abundance of all transcripts analysed in the immature oocyte. During the following stages of preimplantation development, the mRNAs examined exhibited different patterns of expression, but often significant decreases were observed during maturation and maternal–embryonic transition. The transcription of the four genes did not resume with activation of the genome. [ABSTRACT FROM AUTHOR]