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猪卵母细胞体外成熟过程中的能量状态特征受其减数分裂能力的影响。

Energy Status Characteristics of Porcine Oocytes During In Vitro Maturation is Influenced by Their Meiotic Competence.

作者信息

Milakovic I, Jeseta M, Hanulakova S, Knitlova D, Hanzalova K, Hulinska P, Machal L, Kempisty B, Antosik P, Machatkova M

机构信息

Department of Genetics and Reproduction, Veterinary Research Institute, Brno, Czech Republic.

Department of Animal Breeding, Faculty of Agronomy, Mendel University in Brno, Brno, Czech Republic.

出版信息

Reprod Domest Anim. 2015 Oct;50(5):812-9. doi: 10.1111/rda.12592. Epub 2015 Aug 17.

Abstract

The characteristics of energy status in porcine oocytes as related to their meiotic competence and in vitro maturation were studied. Cycling pubertal gilts in the early luteal to early follicular phases of the ovarian cycle were used as oocyte donors. The oocytes recovered from medium (MF) or small follicles (SF) were considered meiotically more or less competent, respectively. A half of oocytes from each category was matured by the standard protocol. The oocytes were examined before or after maturation by confocal microscopy, a bioluminescent cell assay and Western blotting. Four experiments, each in triplicate, were performed to assess both SF and MF oocytes in terms of metabolic units formed by mitochondria and lipids, ATP and lipid consumption and lipid droplets with adipose differentiation-related protein (ADRP) expression. The proportion of oocytes with metabolic units, the mean ATP content and the number of lipid droplets per oocyte, and the relative number of lipid droplets with ADRP expression were significantly higher in the MF compared to SF oocytes before maturation. On the other hand, after maturation, there was an increase in the proportion of oocytes with metabolic units and the relative number of lipid droplets with ADRP expression in the SF compared to MF oocytes. In conclusion, specific differences in energy characteristics between porcine oocytes with different meiotic competence were found. Meiotically more competent oocytes are more advanced in terms of energy reserves before maturation, while meiotically less competent oocytes are more active in replenishing energy stores during maturation.

摘要

研究了猪卵母细胞的能量状态特征及其与减数分裂能力和体外成熟的关系。处于卵巢周期黄体早期至卵泡早期的青春期发情母猪被用作卵母细胞供体。从中等大小卵泡(MF)或小卵泡(SF)中回收的卵母细胞分别被认为减数分裂能力或多或少。每类卵母细胞的一半通过标准方案进行成熟培养。在成熟前后,通过共聚焦显微镜、生物发光细胞测定和蛋白质免疫印迹法对卵母细胞进行检测。进行了四个实验,每个实验重复三次,以评估SF和MF卵母细胞在线粒体和脂质形成的代谢单位、ATP和脂质消耗以及与脂肪分化相关蛋白(ADRP)表达的脂滴方面的情况。在成熟前,与SF卵母细胞相比,MF卵母细胞中具有代谢单位的卵母细胞比例、平均ATP含量、每个卵母细胞的脂滴数量以及具有ADRP表达的脂滴相对数量显著更高。另一方面,在成熟后,与MF卵母细胞相比,SF卵母细胞中具有代谢单位的卵母细胞比例和具有ADRP表达的脂滴相对数量增加。总之,发现了具有不同减数分裂能力的猪卵母细胞在能量特征方面的特定差异。减数分裂能力较强的卵母细胞在成熟前的能量储备方面更先进,而减数分裂能力较弱的卵母细胞在成熟过程中补充能量储存更为活跃。

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