Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, via Morego 30, 16163 Genova, Italy.
Nat Commun. 2012 Jul 17;3:960. doi: 10.1038/ncomms1961.
In utero electroporation is a powerful tool to transfect and manipulate neural-precursor cells of the rodent parietal cortex and their progeny in vivo. Although this technique can potentially target numerous brain areas, reliability of transfection in some brain regions is low or physical access is limited. Here we present a new in utero electroporation configuration based on the use of three electrodes, the relative position and polarities of which can be adjusted. The technique allows easy access and exceedingly reliable monolateral or bilateral transfection at brain locations that could only be sporadically targeted before. By improvement in the efficiency of the electrical field distribution, demonstrated here by a mathematical simulation, the multi-electrode configuration also extends the developmental timeframe for reliable in utero electroporation, allowing for the first time specific transfection of Purkinje cells in the rat cerebellum.
子宫内电穿孔是一种强大的工具,可在体内转染和操作啮齿动物顶皮质层的神经前体细胞及其后代。尽管该技术有可能靶向许多脑区,但在某些脑区的转染可靠性低或物理接触受限。在这里,我们提出了一种新的基于使用三个电极的子宫内电穿孔配置,其相对位置和极性可以调节。该技术允许在以前只能零星靶向的脑区进行易于接近和极其可靠的单侧或双侧转染。通过本文中的数学模拟证明了电场分布效率的提高,多电极配置还扩展了可靠子宫内电穿孔的发育时间框架,首次允许在大鼠小脑浦肯野细胞中进行特异性转染。