Use of in vitro electroporation and slice culture for gene function analysis in the mouse embryonic spinal cord

Abstract

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.

Keywords: Electroporation; Mouse embryo; Spinal cord; Tissue culture.