Methodology
Rapid and efficient genetic manipulation of gyrencephalic carnivores using in utero electroporation
1 Department of Molecular and Systems Neurobiology, Graduate School of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo, 113-0033, Japan
2 Global COE Program "Comprehensive Center of Education and Research for Chemical Biology of the Diseases", The University of Tokyo, Bunkyo-ku, Tokyo, 113-0033, Japan
3 PRESTO, Japan Science and Technology Agency, Chiyoda-ku, Tokyo, 102-0076, Japan
Molecular Brain 2012, 5:24 doi:10.1186/1756-6606-5-24
Published: 20 June 2012Abstract
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.
