Hwang In-Sun, Park Mi-Rung, Moon Hyo-Jin, Shim Joo-Hyun, Kim Dong-Hoon, Yang Byoung-Chul, Ko Yeoung-Gyu, Yang Boh-Suk, Cheong Hee-Tae, Im Gi-Sun
Division of Animal Biotechnology, National Livestock Research Institute, RDA, Suwon, Korea.
Mol Reprod Dev. 2008 Mar;75(3):464-71. doi: 10.1002/mrd.20785.
This study examined whether high osmolarity of culture medium at the early culture stage affects development and expression of apoptosis related genes (Bax-alpha and Bcl-xl) of porcine nuclear transfer (NT) and in vitro fertilization (IVF) embryos. NT and IVF embryos were divided into three groups and the basic medium was PZM-3 (260-270 mOsmol, control group). The control group of embryos was cultured in PZM-3 for whole culture period. Other two groups of embryos were cultured in a modified PZM-3 with 0.05 M sucrose (300-320 mOsmol, sucrose group) or increased NaCl to 138 mM (300-320 mOsmol, NaCl group) for the first 2 days, and then cultured in PZM-3 for 4 days. NT embryos cultured in NaCl group showed a significantly higher developmental rate to the blastocyst stage with a decreased apoptosis rate compared to the control (P < 0.05). There was no difference in blastocyst formation and apoptosis incidence among the three culture treatments for IVF-derived embryos. Bax-alpha mRNA expression was significantly higher in the control than sucrose or NaCl group for both NT and IVF embryos (P < 0.05). Moreover, the relative abundance of Bax-alpha/Bcl-xl was higher in the control than the treatment groups. These results indicate that the higher osmolarity at the early embryonic stage of porcine NT and IVF embryos can improve the in vitro development with reduced apoptosis through regulating the Bax-alpha/Bcl-xl gene expression.
本研究检测了早期培养阶段培养基的高渗透压是否会影响猪核移植(NT)胚胎和体外受精(IVF)胚胎的发育以及凋亡相关基因(Bax-α和Bcl-xl)的表达。将NT胚胎和IVF胚胎分为三组,基础培养基为PZM-3(260 - 270 mOsmol,对照组)。对照组胚胎在整个培养期均培养于PZM-3中。另外两组胚胎在最初2天培养于添加0.05 M蔗糖的改良PZM-3(300 - 320 mOsmol,蔗糖组)或添加NaCl至138 mM的改良PZM-3(300 - 320 mOsmol,NaCl组)中,然后在PZM-3中培养4天。与对照组相比,在NaCl组中培养的NT胚胎发育至囊胚阶段的发育率显著更高,凋亡率降低(P < 0.05)。对于IVF来源的胚胎,三种培养处理之间的囊胚形成和凋亡发生率没有差异。对于NT胚胎和IVF胚胎,对照组中Bax-α mRNA表达均显著高于蔗糖组或NaCl组(P < 0.05)。此外,对照组中Bax-α/Bcl-xl的相对丰度高于处理组。这些结果表明,猪NT胚胎和IVF胚胎早期胚胎阶段的较高渗透压可通过调节Bax-α/Bcl-xl基因表达来改善体外发育并减少凋亡。