Kumar Manish, Faraji Mona, Sarwalia Parul, Kumar Sandeep, Gohain Moloya, De Sachinandan, Kumar Rakesh, Datta T K
1Animal Genomics Laboratory,Animal Biotechnology Centre,National Dairy Research Institute,Karnal - 132001HaryanaIndia.
Zygote. 2018 Oct;26(5):359-365. doi: 10.1017/S0967199418000321. Epub 2018 Oct 5.
SummaryMaturing oocytes have diverse developmental potential and good quality oocytes exhibit a better ability to attain physiological milestones in a time-dependent manner. This situation necessitates the confirmation of oocyte developmental status more precisely under an in vitro embryo production (IVEP) regime. The aim of this study was to explain timely events in germinal vesicle breakdown (GVBD), an important milestone of oocyte nuclear maturation, to delineate the developmental capacity of Bubalus bubalis oocytes. In addition, the expression profile of genes responsible for GVBD was assessed in order to understand the molecular context responsible for GVBD. The chronology of GVBD events at different time intervals during in vitro maturation (IVM) suggests that the rate at which oocytes undergo GVBD was strikingly different in the brilliant cresyl blue (BCB)+ and BCB- groups. The expression of AKT and CDC25B genes for BCB+ oocytes was maximum at 8 h of IVM, and CCNB (cyclin B) peaked at around 10 h, which suggested that GVBD was finished after 10 h in BCB+ oocytes, whereas the expression of AKT and CDC25B was found to peak at around 12-14 h of IVM. This difference consequently delays the GVBD event by 2-4 h in BCB- oocytes. Poor abundance of gene transcripts was mainly implicated in delay and lower rate of GVBD in BCB- oocytes which in turn strongly affected the translational ability of oocytes to blastocysts. The findings of this study support the idea that there is a propensity in sub-optimal grade oocytes for delayed GVBD that compromises the developmental ability of low grade buffalo oocytes. The study highlights the very small, but importantly vital and separate, time window of the GVBD event during which the competence levels of buffalo oocytes are altered along with their translational ability to develop into the prospective embryos.
摘要
成熟的卵母细胞具有多种发育潜能,优质卵母细胞更有能力以时间依赖的方式达成生理发育阶段。这种情况使得在体外胚胎生产(IVEP)体系下更精确地确认卵母细胞发育状态成为必要。本研究旨在阐释生发泡破裂(GVBD)这一卵母细胞核成熟的重要阶段中的适时事件,以描绘水牛卵母细胞的发育能力。此外,评估了负责GVBD的基因的表达谱,以了解GVBD背后的分子机制。体外成熟(IVM)期间不同时间间隔的GVBD事件时间顺序表明,灿烂甲酚蓝(BCB)阳性和BCB阴性组的卵母细胞经历GVBD的速率显著不同。BCB阳性卵母细胞中AKT和CDC25B基因的表达在IVM 8小时时最高,CCNB(细胞周期蛋白B)在约10小时达到峰值,这表明BCB阳性卵母细胞在10小时后完成GVBD,而AKT和CDC25B的表达在IVM约12 - 14小时达到峰值。这种差异使BCB阴性卵母细胞的GVBD事件延迟2 - 4小时。基因转录本丰度低主要与BCB阴性卵母细胞中GVBD延迟和速率降低有关,进而严重影响卵母细胞发育为囊胚的转化能力。本研究结果支持这样的观点,即次优等级的卵母细胞存在GVBD延迟的倾向,这损害了低等级水牛卵母细胞的发育能力。该研究突出了GVBD事件中非常小但至关重要且独立的时间窗口,在此期间水牛卵母细胞的能力水平及其发育为预期胚胎的转化能力会发生改变。