Molecular Genetics Institute, Russian Academy of Sciences, Moscow;
Acta Naturae. 2009 Apr;1(1):109-14.
The influence that the expression of the human (glial-derived neurotrophic factor (GDNF)) neurotrophic factor has on the morphology and proliferative activity of embryonic stem cells (SC) of a mouse with R1 lineage, as well as their ability to form embroid bodies (EB), has been studied. Before that, using a PCR (polymerase chain reaction) coupled with reverse transcription, it was shown that, in this very lineage of the embryonic SC, the expression of the receptors' genes is being fulfilled for the neurotropfic RET and GFRα1 glia factor. The mouse's embryonic SC lineage has been obtained, transfected by the human GDNF gene, and has been fused with the "green" fluorescent protein (GFP) gene. The presence of the expression of the human GDNF gene in the cells was shown by northern hybridization and the synthesis of its albuminous product by immunocitochemical coloration with the use of specific antibodies. The reliable slowing-down of the embriod-body formation by the embryonic SC transfected by the GDNF gene has been shown. No significant influence of the expression of the GDNF gene on the morphology and the proliferative activity of the transfected embryonic SCs has been found when compared with the control ones.
研究了人（胶质细胞源性神经营养因子（GDNF））神经营养因子的表达对 R1 系小鼠胚胎干细胞（SC）的形态和增殖活性以及它们形成胚状体（EB）的能力的影响。在此之前，通过聚合酶链反应（PCR）与反转录相结合，表明在这种胚胎 SC 谱系中，神经营养 RET 和 GFRα1 胶质因子的受体基因表达得到了满足。获得了经人 GDNF 基因转染的小鼠胚胎 SC 系，并与“绿色”荧光蛋白（GFP）基因融合。通过 northern 杂交显示了细胞中人 GDNF 基因的表达，并通过使用特异性抗体进行免疫细胞化学染色显示其白蛋白产物的合成。结果表明，转染 GDNF 基因的胚胎 SC 形成胚状体的速度明显减慢。与对照组相比，未发现 GDNF 基因的表达对转染的胚胎 SC 的形态和增殖活性有显著影响。
