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Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation
Authors:Roberta Haddad-Tóvolli  Nora-Em?ke Szabó  Xunlei Zhou  Gonzalo Alvarez-Bolado
Institution:1.Institute of Anatomy and Cell Biology, University of Heidelberg;2.Institut de recherches cliniques de Montreal
Abstract: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.
Keywords:Neuroscience  Issue 77  Neurobiology  Genetics  Cellular Biology  Molecular Biology  Biomedical Engineering  Developmental Biology  Anatomy  Physiology  Embryo  Mammalian  Brain  Diencephalon  Hypothalamus  Genetic Techniques  Transfection  anesthesia  development  electrodes  electroporation    in utero  mammillary body  mouse  animal model
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