Name: In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos (Video) | JoVE Journal
Uploaded: 2007-07-19T00:00:00.000+00:00
Duration: 19 min 4 s
Description: Learn in utero intraventricular injection and electroporation in E16 rat embryos for developmental neuroscience research. This method enables genetic manipulation in a model where traditional techniques are challenging.

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in utero intraventricular injection and electroporation of e16 rat embryos

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.

In-utero in-vivo injection and electroporation of the embryonic rat neocortex provides a powerful tool for the manipulation of individual progenitors ...

Hi, my name is William Ti.I'm from the Arnold Creek Stein Lab. Today, myself and others are gonna be showing you intrauterine injection of E 16 rat embryos. Hi, my name is Laura Elias.Today I'm gonna be showing you how to prepare the microinjection pipettes and load the pipettes for the int ventricular injections and electroporation that we will be doing today. I'll also be showing you how to perform the interventricular injections and electroporation in E 16 rat embryos. So in utero, eject injections and electroporation in the rat are particularly useful because the red is a system where you cannot easily do genetic Manipulations.Okay, so there's a number Of surgical tools that we use for this procedure. We use a freer, a couple of addin forceps, one, one pair that's serrated, and another pair that has a little tooth on the end. We use a pair of hemostats.This is a pair of scissors, and then we use these two staplers. This is the larger ones that, that we use for rats. This is the smaller one that we use for mice, uses a size seven staple, and this is used as a size nine staple and they're just spring loaded.Some of the things that we use here to assist with the surgeries, we just

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JoVE Journal

Biology

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Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse

Periventricular nodular heterotopia (PNH) is the most common form of malformation of cortical development (MCD) in adulthood but its genetic basis remains unknown in most sporadic cases. We have recently developed a strategy to identify novel candidate genes for MCDs and to directly confirm their causative role in vivo.

A Novel Strategy Combining Array-CGH, Whole-exome Sequencing and In Utero Electroporation in Rodents to Identify Causative Genes for Brain Malformations

Learn in utero intraventricular injection and electroporation in E16 rat embryos for developmental neuroscience research. This method enables genetic manipulation in a model where traditional techniques are challenging.

In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos

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