Rapid and efficient genetic manipulation of gyrencephalic carnivores using in utero electroporation.
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.


CONCLUSIONS
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.
Department of Molecular and Systems Neurobiology, Graduate School of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan. kawasaki@m.u-tokyo.ac.jp
Hiroshi
H
Kawasaki
Lena
L
Iwai
Kaori
K
Tanno
D000818:Animals; D000831:Animals, Newborn; D060435:Carnivory; D002540:Cerebral Cortex / Q000502:physiology*; D018274:Electroporation / Q000379:methods*; D005260:Female; D005289:Ferrets / Q000235:genetics*; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D011247:Pregnancy; D013997:Time Factors; D014599:Uterus / Q000378:metabolism*
D049452:Green Fluorescent Proteins