In utero intraventricular injection and electroporation of E16 rat embryos.
In-utero in-vivo injection and electroporation of the embryonic rat neocortex provides a powerful tool for the manipulation of individual progenitors lining the walls of the lateral ventricle. This technique is now widely used to study the processes involved in corticogenesis by over-expressing or knocking down genes and observing the effects on cellular proliferation, migration, and differentiation. In comparison to traditional knockout strategies, in-utero electroporation provides a rapid means to manipulate a population of cells during a specific temporal window. In this video protocol we outline the experimental methodology for preparing rats for surgery, exposing the uterine horns through laporatomy, injecting DNA into the lateral ventricles of the developing embryo, electroporating DNA into the progenitors lining the lateral wall, and caring for animals post-surgery. Our laboratory uses this protocol for surgeries on E15-E21 rats, however it is most commonly performed at E16 as shown in this video.
Walantus
William
W
Institute for Regeneration Medicine, University of California, San Francisco, USA. walantusw@stemcell.ucsf.edu
Elias
Laura
L
Kriegstein
Arnold
A
D000818:Animals; D004247:DNA / Q000008:administration & dosage* / Q000493:pharmacokinetics; D018274:Electroporation / Q000379:methods*; D005260:Female; D005865:Gestational Age; D007276:Injections, Intraventricular / Q000379:methods*; D007813:Laparotomy; D020547:Lateral Ventricles / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D011247:Pregnancy; D051381:Rats / Q000196:embryology*; D013234:Stem Cells / Q000378:metabolism; D014599:Uterus
D004247:DNA