The spinal cord is an important part of the central nervous system (CNS). At present, the expression of the exogenous gene in the spinal cord of the embryonic mouse needs in utero spinal cord electroporation, but the success rate of this technique is very low. In this study, we have demonstrated the expression of an exogenous gene on one side of the spinal cord by combining two methods-in vitro electroporation of embryonic mouse spinal cord and organ spinal cord slices culture. We took 12-day embryonic mice, injected the green fluorescent protein (pCAGGS-GFP) plasmid into the spinal cord cavity in vitro, and then electroporated. The spinal cord was cut into 300-μm slices using a vibratory microtome. After cultured for 48 h, GFP-positive neurons were clearly observed on one side of the spinal cord, indicating that the exogenous gene was successfully transferred. The axon projection direction is basically unanimous from the inside to the lateral edge of the spinal cord. Compared to neurons in vivo, a single neuron in the culturing section has more complete neurites and is conducive to studying changes in the structure and behavior of individual neurons. Based on the above results, we have successfully established a convenient and efficient method for expressing the exogenous gene in the spinal cord of the mouse.

中文翻译：

---

体外电穿孔和切片培养在小鼠胚胎脊髓基因功能分析中的应用

脊髓是中枢神经系统 (CNS) 的重要组成部分。目前，外源基因在胚胎小鼠脊髓中的表达需要宫内脊髓电穿孔，但该技术的成功率很低。在这项研究中，我们通过结合两种方法 - 胚胎小鼠脊髓的体外电穿孔和器官脊髓切片培养证明了外源基因在脊髓一侧的表达。我们取了12天的胚胎小鼠，将绿色荧光蛋白（pCAGGS-GFP）质粒体外注射到脊髓腔中，然后进行电穿孔。使用振动切片机将脊髓切成 300 微米的切片。培养 48 h 后，脊髓一侧可见 GFP 阳性神经元，说明外源基因转移成功。轴突投射方向从脊髓内侧向外侧缘基本一致。与体内神经元相比，培养切片中的单个神经元具有更完整的神经突，有利于研究单个神经元的结构和行为变化。基于以上结果，我们成功建立了一种方便高效的外源基因在小鼠脊髓中表达的方法。