Spatiotemporal Molecular Approach of in utero Electroporation to Functionally Decipher Endophenotypes in Neurodevelopmental Disorders

Abstract

We have only just begun to decipher the complexity of our brain, including its maturation. Correct brain development and communication among brain areas are crucial for proper cognitive behavior. Brain area-specific genes expressed within a particular time window direct neurodevelopmental events such as proliferation, migration, axon guidance, dendritic arborization, and synaptogenesis. These genes can pose as susceptibility factors in neurodevelopmental disorders eventually resulting in area-specific cognitive deficits. Therefore, in utero electroporation (IUE)-mediated gene transfer can aid in creating valuable animal models in which the regionality and time of expression can be restricted for the targeted gene(s). Moreover, through the use of cell-type-specific molecular constructs, expression can be altered in a particular neuronal subset within a distinct area such that we are now able to causally link the function of that gene in that brain region to the etiology of the disorder. Thus, IUE-mediated gene transfer is an attractive molecular technique to spatiotemporally address the developmental aspects of gene function in relation to neurodevelopmental disorder-associated endophenotypes.

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using <em>In utero</em> Electroporation

“…In addition, the same kind of protocol could be implemented at other stages of embryonic development to target different populations. For example, a developmentally very late cortical electroporation paradigm at E18.5 can be performed to drive expression in astrocytic progenitors 1 . Similarly, while an electroporation of the hippocampus at E14.5 allows to target CA1-CA3 pyramidal neuron progenitors and dentate granule cell progenitor at the same time, a late hippocampal electroporation (E18.5 or early postnatal) would allow to target different dentate granule progenitors 14 .…”

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AbstractIn utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system [1][2][3][4][5] . To date this tool has been widely used to study the regulation of cellular proliferation, differentiation and neuronal migration especially in the developing cerebral cortex [6][7][8] .

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C57BL/6-specific conditions for efficient in utero electroporation of the central nervous system

“…This technique can even be expanded to postnatal stages (Boutin et al, 2008). Different areas of the mouse brain can be manipulated by varying the position of the positive electrode (Kolk et al, 2011;Langevin et al, 2007;Navarro-Quiroga et al, 2007;Saito and Nakatsuji, 2001). The combination of in utero electroporation with transgenic mouse strains additionally enhances the number of possible applications for elucidating the molecular mechanisms of brain development and function (Trimbuch et al, 2009).…”

“…In utero electroporation is a fast an efficient method of specifically addressing gene expression in the murine brain (Fukuchi-Shimogori and Grove, 2001;Saito and Nakatsuji, 2001;Tabata and Nakajima, 2001). In utero electroporation allows for the localized insertion of particular genes and siRNAs into specified brain regions and is therefore a powerful technique for studying the development and function of the brain (Kolk et al, 2011;Nishimura et al, 2012;Saito and Nakatsuji, 2001). Transfected progenitors and their descendant cells express the inserted genes, which enable observation of the resulting cell morphology, proliferation and migration at later stages (Tabata and Nakajima, 2008).…”

MicroRNA-338 modulates cortical neuronal placement and polarity

“…44,45 Timed pregnant (E14.5) mice were anaesthetized with 100 mg/kg Ketamine and 10 mg/kg Xylazine. Following laparotomy, a solution containing the plasmid DNA (2 mg/ml of vector DNA of interest co-injected with pEBFP-N1 (Clontech, Mountain View, CA) in Tris-buffered [pH 7.4] 0.02% Fast Green was injected through the uterine wall into the lateral ventricle of each embryo.…”