Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior
Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior
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DOI：
10.1073/pnas.0308560101
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发表时间：
2004-04-20
影响因子：
11.1
通讯作者：
Gutiérrez-Adán, A
中科院分区：
文献类型：
--
作者：
Fernández-Gonzalez, R;Moreira, P;Gutiérrez-Adán, A
哺乳动物胚胎培养的长期发育和行为后果尚不清楚。通过添加FCS改变培养基，我们想确定在次优条件下培养的小鼠胚胎在胚泡阶段是否会产生印迹基因的异常mRNA表达，以及成年小鼠的胎儿发育、生长和行为是否受到影响。将从超数排卵的雌性B6 CBAF（1）小鼠获得的单细胞胚胎在K+改良的单纯优化培养基中培养4天，培养基中含有10% FCS或1 g/L BSA。胚胎移植后，出生的动物接受了几项发育和行为测试。胎牛血清对囊胚中某些印迹基因的mRNA表达有显著影响。断奶前发育的八项指标中的两项和神经运动发育的一些具体指标，如行走活动，在FCS组中延迟。34周后，在FCS存在下体外培养的雌性小鼠的体重显著高于对照组。此外，小鼠的运动活动在5个月和15个月时发生了改变。对20个月大的动物进行解剖病理学和组织学分析，发现一些大器官和病理学增加。我们发现，胚胎培养的小鼠在焦虑方面表现出特定的行为改变，并表现出内隐记忆的缺陷。我们的数据表明，在胚胎发育的着床前阶段，次优的体外培养会不可逆地影响出生后发育、生长和生理的长期编程。
The long-term developmental and behavioral consequences of mammalian embryo culture are unknown. By altering the culture medium with the addition of FCS, we wanted to determine whether mouse embryos cultured under suboptimal conditions develop aberrant mRNA expression of imprinting genes at the blastocyst stage and whether fetal development, growth, and behavior of adult mice are affected. One-cell embryos obtained from superovulated female B6CBAF(1) mice were cultured for 4 days in K+-modified simplex optimized medium in the presence of either 10% FCS or 1 g/liter BSA. After embryo transfer, born animals were submitted to several developmental and behavior tests. The mRNA expression of some imprinting genes was significantly affected in blastocysts cultured in the presence of FCS. Two of the eight measures of preweaning development and some specific measures of neuromotor development, such as the walking activity, were delayed in the group originated with FCS. After 34 weeks, the weight of female mice cultured in vitro in the presence of FCS was significantly higher than controls. in addition, the locomotion activity of mice was altered at 5 and 15 months. Anatomopathological and histological analysis of animals at 20 months of age showed some large organs and an increase in pathologies. We have found that mice derived from embryos cultured with FCS exhibited specific behavioral alterations in anxiety and displayed deficiencies in implicit memories. Our data indicate that long-term programming of postnatal development, growth, and physiology can be affected irreversibly during the preimplantation period of embryo development by suboptimal in vitro culture.