Targeted gene delivery to telencephalic inhibitory neurons by directional in utero electroporation
Targeted gene delivery to telencephalic inhibitory neurons by directional in utero electroporation
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DOI：
10.1016/j.jneumeth.2004.09.027
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发表时间：
2005-04-30
影响因子：
3
通讯作者：
Callaway, EM
中科院分区：
文献类型：
--
作者：
Borrell, V;Yoshimura, Y;Callaway, EM
端脑抑制性神经元起源于神经节隆起，在分化并整合到局部回路之前，沿着切线轨迹迁移到大脑皮层。目前对中间神经元发育和功能的研究受益于基因敲除和转基因小鼠的使用，而没有利用子宫内电穿孔的多功能性。在这里，我们展示了如何在子宫内电穿孔可以被定向到神经节隆起，以特异性靶向中间神经元的基因表达。将GFP编码质粒电穿孔到神经节隆起中导致在发育过程中迁移的中间神经元的选择性标记。在电穿孔动物的成年脑中，标记了各种各样的皮质、海马和嗅球中间神经元。我们还表明，GFP表达的中间神经元可以在成人大脑皮层的活体切片中可视化，在那里它们显示正常的电生理特性。使用急性切片的光刺激研究表明，皮质GFP+中间神经元接收正常的层特异性突触输入，表明这些神经元整合在局部皮质电路中。因此，子宫内神经节隆起定向电穿孔是一种有效、快速和多功能的选择性切除端脑中间神经元的方法，对于发育研究和成人功能研究都是最佳的。(c)2004 Elsevier B.V.保留所有权利。
Telencephalic inhibitory neurons originate in the ganglionic eminences and migrate to the cerebral cortex following a tangential trajectory, before they differentiate and integrate within the local circuitry. Current studies of interneuron development and function benefit from the use of knock-out and transgenic mice, whereas none take advantage of the versatility of in utero electroporation. Here, we show how in utero electroporation can be directed to the ganglionic eminences to specifically target gene expression to interneurons. Electroporation of GFP-encoding plasmids into the ganglionic eminences results in selective labeling of migrating interneurons during development. In the adult brain of electroporated animals, a wide variety of cortical, hippocampal and olfactory bulb interneurons are labeled. We also show that GFP-expressing interneurons can be visualized in living slices of adult cerebral cortex, where they display normal electrophysiological properties. Photostimulation studies using acute slices show that cortical GFP+ interneurons receive normal, layer-specific synaptic input, indicating that these neurons integrate within the local cortical circuitry. Ganglionic eminence-directed in utero electroporation is therefore an effective, rapid, and versatile method of selectively transfecting telencephalic interneurons, optimal for both developmental studies and adult functional studies. (c) 2004 Elsevier B.V. All rights reserved.