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In vivo and in utero electroporation

Electroporation of nerve cells in embryonic slices or whole brain is a powerful tool to study the fate of selected cells with given gene expression or repression. Quick screening of targeting and rescue experiments for critical genes in cell differentiation and activity ranges from short time cellular imaging (time-lapse microscopy) to long-term behavioural adaptation. Transfection of expression vectors encoding the gene of interest is performed by focal electroporation with a Nepagene apparatus applied to organotypic slices or in utero. In the latter case, the DNA solution is injected into the lateral ventricle of isolated embryos within the intact uterine wall of timed-pregnancy mice and each embryo placed between tweezer-type electrodes of the electroporation apparatus.
See Lebrand, Hornung.

Electroporation is also used to facilitate analysis of mechanisms of synaptic transmission. Using electroporation of dextran-coupled dyes, long-range projections (0.5-1 mm) are labelled rapidly and selectively in the slice. This technique sets the ground for the identification and characterization of novel synaptic afferents from distant projection systems. The Lüthi group is currently interested in afferent systems projecting into higher-order thalamus. See Luthi


GAD67-derived GFP+ neurons expressing Dsred2 are observed within the corpus callosum after electroporating ganglionic eminence neuronal precursors



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