Zhang Jin Yu, Jiang Yong, Lin Tao, Kang Jung Won, Lee Jae Eun, Jin Dong Il
Department of Animal Science & Biotechnology, Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon, Korea.
Mol Reprod Dev. 2015 Jan;82(1):66-77. doi: 10.1002/mrd.22447. Epub 2015 Jan 7.
Lysophosphatidic acid (LPA), a member of the phospholipid autacoid family, is present in human follicular fluid. The aim of the present study was to compare the developmental competence of porcine embryos created via in vitro fertilization (IVF) and parthenogenetic activation (PA) in culture medium supplemented with LPA, in comparison with a control group. The effects of LPA on porcine oocyte maturation and pre-implantation embryonic development were also examined. Addition of 10 μM LPA to the oocyte maturation medium significantly increased the proportion of oocytes reaching metaphase I (MI) or metaphase II (MII), and enhanced embryonic developmental potential. When present during oocyte maturation, LPA significantly increased the abundance of phosphorylated ERK1/2 in MI and MII oocytes, showing that LPA enhanced nuclear maturation via activation of the mitogen-activated protein kinase (MAPK) pathway. In addition, Cyclin B1 levels were elevated in MI- and MII-stage oocytes, suggesting that LPA plays a role in both nuclear and cytoplasmic maturation of oocytes. After fertilization, the frequency of polyspermy in embryos obtained using LPA-treated oocytes was less than that in the control group. Further, blastocyst formation and blastocyst cell number were enhanced and apoptosis was reduced upon LPA treatment of embryos created either by IVF and PA. LPA treatment of blastocysts derived by IVF or PA resulted in increased expression of the anti-apoptotic BCL2L1 gene while reducing expression of the pro-apoptotic genes BAX and CASP3. Together, our data indicate that LPA supplementation improves porcine oocyte maturation and subsequent in vitro development of pre-implantation embryos.
溶血磷脂酸（LPA）是磷脂自分泌家族的一员，存在于人类卵泡液中。本研究的目的是比较在添加LPA的培养基中通过体外受精（IVF）和孤雌激活（PA）产生的猪胚胎与对照组相比的发育能力。还研究了LPA对猪卵母细胞成熟和植入前胚胎发育的影响。在卵母细胞成熟培养基中添加10μM LPA可显著提高达到中期I（MI）或中期II（MII）的卵母细胞比例，并增强胚胎发育潜力。当LPA在卵母细胞成熟过程中存在时，它显著增加了MI和MII期卵母细胞中磷酸化ERK1/2的丰度，表明LPA通过激活丝裂原活化蛋白激酶（MAPK）途径增强了核成熟。此外，MI期和MII期卵母细胞中的细胞周期蛋白B1水平升高，表明LPA在卵母细胞的核成熟和细胞质成熟中均发挥作用。受精后，使用LPA处理的卵母细胞获得的胚胎中多精受精的频率低于对照组。此外，对通过IVF和PA产生的胚胎进行LPA处理后，囊胚形成和囊胚细胞数量增加，细胞凋亡减少。对IVF或PA衍生的囊胚进行LPA处理导致抗凋亡BCL2L1基因的表达增加，同时降低促凋亡基因BAX和CASP3的表达。总之，我们的数据表明补充LPA可改善猪卵母细胞成熟以及随后植入前胚胎的体外发育。
