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 JoVE Biology

In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos

1, 1, 1

1Institute for Regeneration Medicine, University of California, San Francisco - UCSF

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    Cite this Article

    Walantus, W., Elias, L., Kriegstein, A. In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos. J. Vis. Exp. (6), e236, doi:10.3791/236 (2007).

    Abstract

    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.

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    16 Comments

    Very nice work. I do this routinely in Mouse but have little luck with Rat. I get poor survival of pups. If I do a sham experiment, I get fine survival with electroporation only but little survival if I inject only. I've used a variety of anesthetics. Any suggestions particular to Rat? Thank you much
    Reply

    Posted by: AnonymousOctober 24, 2007, 2:50 PM

    Try different methods of purifying the DNA.
    Reply

    Posted by: AnonymousOctober 29, 2007, 5:19 PM

    I am having poor survival as well. Any ideas about different methods of purification? I am currently using a Promega MIDI/Maxi kit. Have you noticed with purification methods are better?
    Reply

    Posted by: AnonymousOctober 18, 2010, 1:53 PM

    hi barry. are you beveling your injection pipettes enough? if they are not sharp enough embryos will have high mortality. have a look at the fourth chapter of the video again.
    Reply

    Posted by: Aaron K.October 31, 2007, 2:12 AM

    Thanks, will try, but mice go just fine without problems except that B6 mice don't make the best of moms.
    Reply

    Posted by: AnonymousOctober 31, 2007, 8:53 AM

    Hi, I would appreciate if you could send me the protocol for mice ...Thanks a lot !
    Martine
    m.barkats@institut-myologie.org
    Reply

    Posted by: Martine B.May 28, 2010, 11:40 AM

    Where can we find the tools used to make the syrine injector tip?
    Reply

    Posted by: AnonymousJuly 17, 2008, 2:59 PM

    what are the medicine for motor neuron disease?
    Reply

    Posted by: AnonymousSeptember 8, 2009, 1:23 AM

    From which company can I buy the electrods for the electroporation. The same as you have. Could you tell me their cat. # also?
    Reply

    Posted by: AnonymousNovember 10, 2009, 4:19 PM


    BTX Harvard Apparatus Tweezertrodes, you can get them from Fisher and other vendors. -William
    Reply

    Posted by: AnonymousNovember 10, 2009, 4:40 PM

    I was not able to find the volume for these intraventricular infusions. Also, what is the maximum infusion volume for these kinds of procedures using newborn rats? Thanks!
    Reply

    Posted by: mark b.August 7, 2010, 8:49 AM

    I was not able to find the volume for these intraventricular infusions. Also, what is the maximum infusion volume for these kinds of procedures using newborn rats? Thanks!
    Reply

    Posted by: mark b.August 7, 2010, 8:50 AM

    Have you ever delivered DNA earlier into the lateral ventricles (E1².5 or E13)?

    Thank you.

    Shelley
    Reply

    Posted by: AnonymousNovember 30, 2011, 11:22 PM

    No. Sorry, There are some who do though. You can use ultrasound or IR videomicroscopy.
    Reply

    Posted by: AnonymousDecember 1, 2011, 4:17 PM

    Thank you...any one in particular that you know has a technique working or a paper published? Thanks!
    Reply

    Posted by: AnonymousDecember 1, 2011, 4:56 PM

    Thank you...any one in particular that you know has a technique working or a paper published? Thanks!
    Reply

    Posted by: AnonymousDecember 1, 2011, 4:57 PM

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