Tanaka Hiroshi, Takeo Shun, Abe Takahito, Kin Airi, Shirasuna Koumei, Kuwayama Takehito, Iwata Hisataka
Tokyo University of Agriculture, Kanagawa 243-0034, Japan.
J Reprod Dev. 2016 Jun 17;62(3):235-40. doi: 10.1262/jrd.2015-143. Epub 2016 Jan 30.
The aim of the present study was to examine the fertilization ability and mitochondrial function of oocytes derived from cows with or without liver damage. Oocytes were collected from the ovaries of cows with damaged livers (DL) and those of cows with healthy livers (HL), subjected to in vitro maturation, and fertilized in vitro. A significantly high abnormal fertilization rate was observed for oocytes from DL cows compared to oocytes from HL cows. The time to dissolve the zona pellucida by protease before fertilization was similar between the two liver conditions, whereas after fertilization treatment this time was shorter for DL cows than for HL cows. The percentage of oocytes with equivalent cortical granule distributions underneath the membrane was greater for in vitro matured oocytes from HL cows, whereas an immature distribution pattern was observed for oocytes from DL cows. In addition, a greater percentage of oocytes derived from HL cows released cortical granules following fertilization compared with oocytes from DL cows. Mitochondrial function determined by ATP content and membrane potential were similar at the germinal vesicle stage, but post-in vitro maturation, the oocytes derived from HL cows showed higher values than DL cows. The mitochondrial DNA copy number in oocytes was similar between the two liver conditions for both the germinal vesicle and post-in vitro maturation oocytes. In conclusion, liver damage induces low fertilization, likely because of incomplete cortical granule distribution and release, and the maturation of oocytes from DL cows contain low-functioning mitochondria compared to their HL counterparts.
本研究的目的是检测来自有或没有肝脏损伤奶牛的卵母细胞的受精能力和线粒体功能。从肝脏受损奶牛(DL)和肝脏健康奶牛(HL)的卵巢中采集卵母细胞,进行体外成熟,并进行体外受精。与HL奶牛的卵母细胞相比,DL奶牛的卵母细胞观察到显著较高的异常受精率。两种肝脏状况下,受精前蛋白酶溶解透明带的时间相似,而受精处理后,DL奶牛的这一时间比HL奶牛短。HL奶牛体外成熟卵母细胞膜下皮质颗粒分布相同的卵母细胞百分比更高,而DL奶牛的卵母细胞则观察到未成熟的分布模式。此外,与DL奶牛的卵母细胞相比,HL奶牛的卵母细胞在受精后释放皮质颗粒的百分比更高。在生发泡期,由ATP含量和膜电位测定的线粒体功能相似,但体外成熟后,HL奶牛的卵母细胞的值高于DL奶牛。对于生发泡期和体外成熟后的卵母细胞,两种肝脏状况下卵母细胞中的线粒体DNA拷贝数相似。总之,肝脏损伤导致受精率低,可能是由于皮质颗粒分布和释放不完全,并且与HL奶牛的卵母细胞相比,DL奶牛的卵母细胞成熟时线粒体功能较低。