Xu Mingzhu, Zeng Yaqiong, Chi Daming, Si Linan, Qu Xiao, Li Juan
College of Animal Science and Technology,Nanjing Agricultural University,Nanjing Weigang No. 1,Jiangsu Province 210095,People's Republic of China.
Zygote. 2018 Feb;26(1):40-49. doi: 10.1017/S0967199417000697. Epub 2017 Dec 13.
Lipid droplets (LDs) are the main energy resource for porcine preimplantation embryonic development. PLIN3 has been implicated in LD formation and regulation. Therefore, this study aimed to detect the dynamic pattern of PLIN3 in pig oocytes and cumulus cells (CC) during in vitro maturation (IVM), and to determine the relationship between PLIN3 and LD content. IVM with cumulus-enclosed oocytes (CEO), cumulus-denuded oocytes (DO) and the CCs denuded from the corresponding oocytes (DCC) was performed in porcine follicular fluid (PFF) or PFF-free optimized medium. DO and the DCC were cultured together under the same conditions as described above, while the DO was named DTO and the DCC was named DTCC in this group. Firstly, our results revealed LDs distributed widely in oocytes and CC, while the PLIN3 protein coated these LDs and spread out ubiquitously in the cytoplasm. Secondly, not only the mRNA level but also at protein level of PLIN3 in immature naked oocytes (IO) was higher than that in matured CEO, DO and DTO. Although PLIN3 was expressed at lower levels in CC from immature oocytes (ICC), the protein level of PLIN3 was comparably higher in the ECC and DCC groups. The triglyceride (TG) content in CEO and DO was significantly less abundant compared with that in IO. Therefore, our results indicated that co-culturing of oocytes and CC might affect PLIN3 expression levels in CC but not in oocytes. Lipid accumulation in pig oocytes during maturation might be affected by PLIN3 cross-talk between oocytes and CC.
脂滴(LDs)是猪植入前胚胎发育的主要能量来源。PLIN3与脂滴的形成和调控有关。因此,本研究旨在检测猪卵母细胞和卵丘细胞(CC)在体外成熟(IVM)过程中PLIN3的动态模式,并确定PLIN3与脂滴含量之间的关系。在猪卵泡液(PFF)或无PFF的优化培养基中对卵丘细胞包裹的卵母细胞(CEO)、裸卵(DO)以及从相应卵母细胞中剥除的卵丘细胞(DCC)进行IVM。DO和DCC在与上述相同条件下共同培养,在该组中DO命名为DTO,DCC命名为DTCC。首先,我们的结果显示脂滴广泛分布于卵母细胞和CC中,而PLIN3蛋白包裹这些脂滴并在细胞质中普遍分布。其次,未成熟裸卵(IO)中PLIN3的mRNA水平和蛋白水平均高于成熟的CEO、DO和DTO。虽然PLIN3在未成熟卵母细胞的CC(ICC)中表达水平较低,但在ECC和DCC组中PLIN3的蛋白水平相对较高。与IO相比,CEO和DO中的甘油三酯(TG)含量明显较少。因此,我们的结果表明卵母细胞和CC共同培养可能会影响CC中PLIN3的表达水平,但不会影响卵母细胞中的表达水平。猪卵母细胞成熟过程中的脂质积累可能受到卵母细胞与CC之间PLIN3相互作用的影响。