ABSTRACT In utero electroporation (IUE) is a useful technique for gene delivery in embryonic mouse brain. IUE technique is used to investigate the mammalian brain development in vivo. However, according to recent studies, IUE methodology has some limitations like the formation of artificial ectopias and heterotopias at the micro-injection site. Thus far, the artificial heterotopias generated by physical trauma during IUE are rarely reported. Here, we reported the artificial heterotopias and ectopias generated from surgical damages of micropipette in detail, and moreover, we described the protocol to avoid these phenotypes. For the experimental purpose, we transferred empty plasmids (pCAGIG-GFP) with green fluorescent-labelled protein into the cortical cortex by IUE and then compared the structure of the cortex region between the injected and un-injected cerebral hemispheres. The coronary section showed that ectopias and heterotopias were appeared on imperfect-injected brains, and layer maker staining, which including Ctip2 and TBR1 and laminin, can differentiate the physical damage, revealing the neurons in artificial ectopic and heterotopic area were not properly arranged. Moreover, premature differentiation of neurons in ectopias and heterotopias were observed. To avoid heterotopias and ectopias, we carefully manipulated the method of IUE application. Thus, this study might be helpful for the in utero electroporator to distinguish the artificial ectopias and heterotopias that caused by the physical injury by microneedle and the ways to avoid those undesirable circumstances.

中文翻译：

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子宫电穿孔过程中手术损伤引起的小鼠新皮质神经细胞迁移中异位的识别和预防

摘要 子宫内电穿孔 (IUE) 是一种在小鼠胚胎脑中进行基因传递的有用技术。IUE 技术用于研究哺乳动物的体内大脑发育。然而，根据最近的研究，IUE 方法有一些局限性，如在显微注射部位形成人工异位和异位。迄今为止，IUE期间身体创伤产生的人工异位很少见报道。在这里，我们详细报告了微量移液器手术损伤产生的人工异位和异位，此外，我们还描述了避免这些表型的协议。出于实验目的，我们通过 IUE 将带有绿色荧光标记蛋白的空质粒 (pCAGIG-GFP) 转移到皮层皮层，然后比较注射和未注射大脑半球之间的皮层区域结构。冠脉切片显示不完全注射的脑组织出现异位和异位，层标记染色，包括Ctip2和TBR1和层粘连蛋白，可以区分物理损伤，揭示人工异位和异位区域的神经元排列不正确。此外，观察到异位和异位中神经元的过早分化。为了避免异位和异位，我们仔细操作了IUE应用的方法。因此，