Genetic manipulation of the mouse developing hypothalamus through In utero electroporation

Genetic modification of specific regions of the developing mammalian brain is a very powerful experimental approach. However, generating novel mouse mutants is often frustratingly slow. It has been shown that access to the mouse brain developing in utero with reasonable post-operatory survival is po...

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Main Authors: Haddad-Tóvolli, Roberta (Author) , Zhou, Xunlei (Author) , Alvarez-Bolado, Gonzalo (Author)
Format: Article (Journal) Video
Language:English
Published: 7/24/2013
In: JoVE. Video journal
Year: 2013, Issue: 77
ISSN:1940-087X
DOI:10.3791/50412
Subjects:
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.3791/50412
Verlag, lizenzpflichtig, Volltext: https://www.jove.com/v/50412/genetic-manipulation-mouse-developing-hypothalamus-through-utero
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Author Notes:Roberta Haddad-Tóvolli, Nora-Emöke Szabó, Xunlei Zhou, Gonzalo Alvarez-Bolado

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520 |a Genetic modification of specific regions of the developing mammalian brain is a very powerful experimental approach. However, generating novel mouse mutants is often frustratingly slow. It has been shown that access to the mouse brain developing in utero with reasonable post-operatory survival is possible. Still, results with this procedure have been reported almost exclusively for the most superficial and easily accessible part of the developing brain, i.e. the cortex. The thalamus, a narrower and more medial region, has proven more difficult to target. Transfection into deeper nuclei, especially those of the hypothalamus, is perhaps the most challenging and therefore very few results have been reported. Here we demonstrate a procedure to target the entire hypothalamic neuroepithelium or part of it (hypothalamic regions) for transfection through electroporation. The keys to our approach are longer narcosis times, injection in the third ventricle, and appropriate kind and positioning of the electrodes. Additionally, we show results of targeting and subsequent histological analysis of the most recessed hypothalamic nucleus, the mammillary body. 
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