158 RELATIONSHIP BETWEEN FOURTH CELL CYCLE DURATION AND POST-TRANSFER VIABILITY IN IN VITRO-FERTILIZED BOVINE EMBRYOS

Abstract

In cattle, the prediction of embryonic viability after embryo transfer is an important research target. A previous study has indicated that the duration of the fourth cell cycle at the time of maternal-zygotic transition, which is involved in in vitro embryonic development, may be an indicator of blastocyst formation; this study showed that embryos with a short fourth cell cycle have a better potential of developing into blastocysts than those with a long fourth cell cycle (Lequarre et al. 2003 Biol. Reprod. 69, 1707–1713). However, the relationship between the fourth cell cycle duration and post-transfer viability of embryos is unclear. The aim of the present study was to examine the effect of the fourth cell cycle duration on embryo development after embryo transfer. Twenty-five IVF bovine embryos were cultured in well-of-the-well culture dishes contained 125 μ of CR1aa supplemented with 5% calf serum at 38.5°C in 5% O₂ and 5% CO₂ for 168 h after insemination. In vitro development of the embryos was monitored using time-lapse cinematography (Sugimura et al. 2010 Biol. Reprod. 83, 970–978). We found that 61% of the blastocysts had a long fourth cell cycle (41.5 ± 5.9 h), which is commonly referred to as the lag phase, whereas the remaining embryos had a short fourth cell cycle (7.4 ± 4.5 h). All the embryos with a short fourth cell cycle exhibited a lag phase in the next cell cycle (32.9 ± 6.6 h). Moreover, embryos with a short fourth cell cycle were found to have a higher blastocyst rate (75.8%) than those with a long fourth cell cycle (48.1%; Student's t-test, P < 0.01). However, embryonic cell number, apoptosis incidence, chromosomal abnormality and O₂ consumption were found to be identical between the 2 groups (Student's t-test, P > 0.05). Real-time reverse-transcription PCR results of the individual blastocysts showed that the relative expression of 5 genes related to pregnancy reorganization, placentation and fetal growth—namely, CDX2, IFN-τ, PLAC8, AKR1B1 and IGF2R—did not differ between the 2 groups (Student's t-test, P > 0.05). Furthermore, blastocysts derived from embryos with long (n = 30) and short (n = 19) fourth cell cycles were transferred into 49 recipient cows; we did not observe any difference between the long and short fourth cell cycles on the rates of pregnancy (long vs short fourth cell cycle, 30.0 vs 52.6%) and delivery (long vs short fourth cell cycle, 30.0 vs 47.4%; Yates' corrected chi-square test, P > 0.10). These results show that blastocysts derived from embryos with either long or short fourth cell cycles have identical developmental competence after embryo transfer. Therefore, the fourth cell cycle duration during maternal-zygotic transition appears to be unavailable as the indicator of post-transfer viability of IVF bovine embryos.

This work was supported by the Research and Developmental Program for New Bio-Industry Initiatives.