Rapid and efficient genetic manipulation of gyrencephalic carnivores using in utero electroporation
Rapid and efficient genetic manipulation of gyrencephalic carnivores using in utero electroporation
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DOI：
10.1186/1756-6606-5-24
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发表时间：
2012-06-20
期刊：
影响因子：
3.6
通讯作者：
Tanno, Kaori
中科院分区：
文献类型：
--
作者：
Kawasaki, Hiroshi;Iwai, Lena;Tanno, Kaori
背景资料：高等哺乳动物如灵长类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，以及大脑皮层的脑回和外侧脑室下区。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，主要是因为可以应用于高等哺乳动物的遗传操作仍然很差。结果：在这里，我们开发并验证了一种快速有效的技术，使基因操作在脑中的gyrencephalic食肉动物使用子宫内电穿孔。转基因表达的雪貂婴儿在电穿孔后几周内获得。GFP表达在胚胎中可检测到，并且在出生后至少2个月观察到。我们的技术是有用的表达转基因在表层和深层皮层神经元，并检查树突状形态和轴突轨迹的GFP表达神经元在雪貂。此外，多个基因有效地共表达在同一neurons.Conclusion：我们的方法有望成为一个强大的工具，调查的基本机制的发展，功能和病理生理学的大脑结构是独特的高等哺乳动物。
Background: 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.Results: 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.Conclusion: 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.