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猪卵泡中卵母细胞染色质状态对体外胚胎发育的影响。

Effect of oocyte chromatin status in porcine follicles on the embryo development in vitro.

作者信息

Lee Joo Bin, Lee Min Gu, Lin Tao, Shin Hyeon Yeong, Lee Jae Eun, Kang Jung Won, Jin Dong-Il

机构信息

Division of Animal and Dairy Science, Chungnam National University, Daejeon 34134, Korea.

出版信息

Asian-Australas J Anim Sci. 2019 Jul;32(7):956-965. doi: 10.5713/ajas.18.0739. Epub 2019 Jan 2.

Abstract

OBJECTIVE

The main goal of this study was to provide a morphological indicator that could be used to select high-quality oocytes of appropriate meiotic and developmental capabilities in pig. The higher quality of immature oocytes, the higher success rates of in vitro maturation (IVM) and in vitro fertilization (IVF). Thus, prior to the IVM culture, it is important to characterize oocytes morphologically and biochemically in order to assess their quality. Two of the largest indicators of oocyte quality are the presence of cumulus cells and status of chromatin. To investigate the effects of porcine oocyte chromatin configurations on the developmental capacity of blastocysts, we assessed oocyte chromatin status according to follicle size and measured the developmental potency of blastocysts.

METHODS

To sort by follicle size, we divided the oocytes into three groups (less than 1 mm, 1 to 3 mm, and more than 3 mm in diameter). To assess chromatin configuration, the oocytes were assessed for their stages (surrounded nucleolus [SN] germinal vesicle [GV], non-surrounded nucleolus [NSN] GV, GV breakdown, metaphase I [MI], pro-metaphase II [proMII], and metaphase II [MII]) at different maturation times (22, 44, and 66 h). To assess the development rate, oocytes of each follicle size were subjected to parthenogenetic activation for further development. Finally, GV oocytes were grouped by their chromatin configuration (SN, SN/NSN, and NSN) and their global transcriptional levels were measured.

RESULTS

SN GV oocytes were more suitable for IVF than NSN GV oocytes. Moreover, oocytes collected from the larger follicles had a greater distribution of SN GV oocytes and a higher developmental capacity during IVM, reaching MII more quickly and developing more often to blastocysts.

CONCLUSION

Porcine oocytes with high-level meiotic and developmental capacity were identified by analyzing the relationship between follicle size and chromatin configuration. The porcine oocytes from large follicles had a significantly higher SN status in which the transcription level was low and could be better in the degree of meiotic progression and developmental capacity.

摘要

目的

本研究的主要目标是提供一种形态学指标,可用于选择具有适当减数分裂和发育能力的优质猪卵母细胞。未成熟卵母细胞质量越高,体外成熟(IVM)和体外受精(IVF)的成功率越高。因此,在IVM培养之前,从形态学和生物化学角度对卵母细胞进行表征以评估其质量非常重要。卵母细胞质量的两个最大指标是卵丘细胞的存在和染色质状态。为了研究猪卵母细胞染色质构型对囊胚发育能力的影响,我们根据卵泡大小评估了卵母细胞的染色质状态,并测量了囊胚的发育潜能。

方法

为了按卵泡大小进行分类,我们将卵母细胞分为三组(直径小于1毫米、1至3毫米和大于3毫米)。为了评估染色质构型,在不同的成熟时间(22、44和66小时)对卵母细胞的阶段(核仁周围 [SN] 生发泡 [GV]、非核仁周围 [NSN] GV、GV 破裂、中期I [MI]、前中期II [proMII] 和中期II [MII])进行评估。为了评估发育率,对每个卵泡大小的卵母细胞进行孤雌激活以进一步发育。最后,将GV卵母细胞按其染色质构型(SN、SN/NSN和NSN)分组,并测量其整体转录水平。

结果

SN GV卵母细胞比NSN GV卵母细胞更适合IVF。此外,从较大卵泡收集的卵母细胞中SN GV卵母细胞的分布更多,并且在IVM期间具有更高的发育能力,更快地达到MII并且更常发育成囊胚。

结论

通过分析卵泡大小与染色质构型之间的关系,鉴定出具有高水平减数分裂和发育能力的猪卵母细胞。来自大卵泡的猪卵母细胞具有明显更高的SN状态,其中转录水平较低,减数分裂进程和发育能力程度可能更好。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a3b/6601069/971f05af624c/ajas-18-0739f1.jpg

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