Wasielak Marta, Więsak Teresa, Bogacka Iwona, Jalali Beenu Moza, Bogacki Marek
Department of Gamete and Embryo Biology,Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences,Tuwima 10,10-747 Olsztyn,Poland.
Department of Gamete and Embryo Biology,Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences,Tuwima 10,10-748 Olsztyn,Poland.
Zygote. 2017 Apr;25(2):120-130. doi: 10.1017/S0967199416000332. Epub 2016 Dec 23.
Maternal effect genes (MEG) play a crucial role in early embryogenesis. In vitro culture conditions may affect MEG expression in porcine oocytes and embryos. We investigated whether in vitro culture medium supplementation with epidermal growth factor (EGF), IL-1β or LIF (leukemia inhibitory factor) affects the mRNA level of ZAR-1 (zygote arrest 1), NPM2 (nucleoplasmin 2) and DPPA3 (developmental associated protein 3) in porcine MII oocytes and embryos. Cumulus-oocyte complexes (COCs) were matured in NCSU-37 medium (control) or in NCSU-37 with EGF 10 ng/ml, IL-1β 10 ng/ml or LIF 50 ng/ml. After maturation for 44-46 h, MII oocytes were preserved for the analysis of MEG mRNA levels (experiment 1). In experiment 2, COCs were fertilized, and the presumptive zygotes were cultured in the same groups. Then, 2-, 4-, 8-cell embryos, morulae and blastocysts were collected for the analysis of MEG mRNA levels. LIF addition to the maturation medium increased MII oocyte numbers (P < 0.05), while EGF and IL-1β did not affect oocyte maturation. Medium supplementation with EGF resulted in lower DPPA3 mRNA levels in MII oocytes and in 2- and 4-cell embryos versus control embryos (P < 0.05). LIF treatment increased DPPA3 mRNA levels in morulae and blastocysts (P < 0.05). Culture with EGF and IL-1β decreased ZAR-1 and NPM2 mRNA levels in 2-cell embryos (P < 0.05). The inclusion of EGF or IL-1β in the porcine in vitro production system influences ZAR-1, NPM2 and DPPA3 mRNA in MII oocytes and embryos but not beyond the 4-cell stage. LIF stimulates oocyte maturation and affects DPPA3 mRNA in porcine morulae and blastocysts in vitro.
母源效应基因(MEG)在早期胚胎发育中起着至关重要的作用。体外培养条件可能会影响猪卵母细胞和胚胎中MEG的表达。我们研究了在体外培养基中添加表皮生长因子(EGF)、白细胞介素-1β(IL-1β)或白血病抑制因子(LIF)是否会影响猪MII期卵母细胞和胚胎中合子阻滞1(ZAR-1)、核质蛋白2(NPM2)和发育相关蛋白3(DPPA3)的mRNA水平。卵丘-卵母细胞复合体(COCs)在NCSU-37培养基(对照)或添加10 ng/ml EGF、10 ng/ml IL-1β或50 ng/ml LIF的NCSU-37培养基中成熟。成熟44 - 46小时后,保存MII期卵母细胞用于分析MEG mRNA水平(实验1)。在实验2中,对COCs进行受精,并将推定的合子在相同组中培养。然后,收集2细胞、4细胞、8细胞胚胎、桑葚胚和囊胚用于分析MEG mRNA水平。在成熟培养基中添加LIF可增加MII期卵母细胞数量(P < 0.05),而EGF和IL-1β不影响卵母细胞成熟。与对照胚胎相比,在培养基中添加EGF会导致MII期卵母细胞以及2细胞和4细胞胚胎中DPPA3 mRNA水平降低(P < 0.05)。LIF处理可增加桑葚胚和囊胚中DPPA3 mRNA水平(P < 0.05)。用EGF和IL-1β培养可降低2细胞胚胎中ZAR-1和NPM2 mRNA水平(P < 0.05)。在猪体外生产系统中添加EGF或IL-1β会影响MII期卵母细胞和胚胎中ZAR-1、NPM2和DPPA3 mRNA水平,但这种影响不会超过4细胞阶段。LIF可刺激卵母细胞成熟,并在体外影响猪桑葚胚和囊胚中的DPPA3 mRNA水平。