title: Induction of protein deletion through in utero electroporation to define deficits in neuronal migration in transgenic models.
abstract: Genetic deletion using the Cre-Lox system in transgenic mouse lines is a powerful tool used to study protein function. However, except in very specific Cre models, deletion of a protein throughout a tissue or cell population often leads to complex phenotypes resulting from multiple interacting mechanisms. Determining whether a phenotype results from disruption of a cell autonomous mechanism, which is intrinsic to the cell in question, or from a non-cell autonomous mechanism, which would result from impairment of that cell's environment, can be difficult to discern. To gain insight into protein function in an in vivo context, in utero electroporation (IUE) enables gene deletion in a small subset of cells within the developing cortex or some other selected brain region. IUE can be used to target specific brain areas, including the dorsal telencephalon, medial telencephalon, hippocampus, or ganglionic eminence. This facilitates observation of the consequences of cell autonomous gene deletion in the context of a healthy environment. The goal of this protocol is to show how IUE can be used to analyze a defect in radial migration in a floxed transgenic mouse line, with an emphasis on distinguishing between the cell autonomous and non-cell autonomous effects of protein deletion. By comparing the phenotype resulting from gene deletion within the entire cortex versus IUE-mediated gene deletion in a limited cell population, greater insight into protein function in brain development can be obtained than by using either technique in isolation.
authors: Department of Cellular and Molecular Medicine, University of Ottawa; ssvob026@uottawa.ca.
authors: Devon S
authors: DS
authors: Svoboda
authors: Department of Cellular and Molecular Medicine, University of Ottawa.
authors: Alysen
authors: A
authors: Clark
authors: Department of Cellular and Molecular Medicine, University of Ottawa.
authors: David S
authors: DS
authors: Park
authors: Department of Cellular and Molecular Medicine, University of Ottawa; rslack@uottawa.ca.
authors: Ruth S
authors: RS
authors: Slack
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D018274:Electroporation / Q000379:methods*; D004622:Embryo, Mammalian; D005260:Female; D017353:Gene Deletion; D055785:Gene Knockdown Techniques / Q000379:methods*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D011247:Pregnancy; D016160:Retinoblastoma Protein / Q000235:genetics / Q000502:physiology*; D050720:Retinoblastoma-Like Protein p107 / Q000235:genetics / Q000502:physiology*
chemical_list: D016160:Retinoblastoma Protein; D050720:Retinoblastoma-Like Protein p107
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