Department of Molecular and Systems Neurobiology, Graduate School of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan.
Mol Brain. 2012 Jun 20;5:24. doi: 10.1186/1756-6606-5-24.
Higher mammals such as primates and carnivores have highly developed unique brain structures such as the ocular dominance columns in the visual cortex, and the gyrus and outer subventricular zone of the cerebral cortex. However, our molecular understanding of the formation, function and diseases of these structures is still limited, mainly because genetic manipulations that can be applied to higher mammals are still poorly available.
Here we developed and validated a rapid and efficient technique that enables genetic manipulations in the brain of gyrencephalic carnivores using in utero electroporation. Transgene-expressing ferret babies were obtained within a few weeks after electroporation. GFP expression was detectable in the embryo and was observed at least 2 months after birth. Our technique was useful for expressing transgenes in both superficial and deep cortical neurons, and for examining the dendritic morphologies and axonal trajectories of GFP-expressing neurons in ferrets. Furthermore, multiple genes were efficiently co-expressed in the same neurons.
Our method promises to be a powerful tool for investigating the fundamental mechanisms underlying the development, function and pathophysiology of brain structures which are unique to higher mammals.
高等哺乳动物（如灵长类动物和食肉动物）具有高度发达的独特脑结构，如视觉皮层中的眼优势柱、大脑皮层的回和外室下带。然而，我们对这些结构的形成、功能和疾病的分子理解仍然有限，主要是因为可应用于高等哺乳动物的遗传操作仍然很少。
我们开发并验证了一种快速有效的技术，可利用子宫内电穿孔对具有脑回的食肉动物进行基因操作。电穿孔后几周内即可获得表达转基因的雪貂幼崽。电转后胚胎中可检测到 GFP 表达，出生后至少 2 个月仍可观察到 GFP 表达。我们的技术可用于在浅层和深层皮质神经元中表达转基因，并可用于检查 GFP 表达神经元的树突形态和轴突轨迹。此外，还可以有效地在同一神经元中共同表达多个基因。
我们的方法有望成为研究高等哺乳动物特有的脑结构发育、功能和病理生理学基础机制的有力工具。
