In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos

Published 7/19/2007
16 Comments
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Biology

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Summary

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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.

Comments

16 Comments

  1. 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: Anonymous
    October 24, 2007 - 2:50 PM
  2. Try different methods of purifying the DNA.

    Reply
    Posted by: Anonymous
    October 29, 2007 - 5:19 PM
  3. 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: Anonymous
    October 18, 2010 - 1:53 PM
  4. 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
  5. Thanks, will try, but mice go just fine without problems except that B6 mice don't make the best of moms.

    Reply
    Posted by: Anonymous
    October 31, 2007 - 8:53 AM
  6. 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
  7. Where can we find the tools used to make the syrine injector tip?

    Reply
    Posted by: Anonymous
    July 17, 2008 - 2:59 PM
  8. what are the medicine for motor neuron disease?

    Reply
    Posted by: Anonymous
    September 8, 2009 - 1:23 AM
  9. 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: Anonymous
    November 10, 2009 - 4:19 PM

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

    Reply
    Posted by: Anonymous
    November 10, 2009 - 4:40 PM
  11. 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
  12. 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
  13. Have you ever delivered DNA earlier into the lateral ventricles (E1².5 or E13)?

    Thank you.

    Shelley

    Reply
    Posted by: Anonymous
    November 30, 2011 - 11:22 PM
  14. No. Sorry, There are some who do though. You can use ultrasound or IR videomicroscopy.

    Reply
    Posted by: Anonymous
    December 1, 2011 - 4:17 PM
  15. Thank you...any one in particular that you know has a technique working or a paper published? Thanks!

    Reply
    Posted by: Anonymous
    December 1, 2011 - 4:56 PM
  16. Thank you...any one in particular that you know has a technique working or a paper published? Thanks!

    Reply
    Posted by: Anonymous
    December 1, 2011 - 4:57 PM

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