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En injection intraventriculaire Utero et l'électroporation d'embryons de rat E16

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

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Cite this Article: En injection intraventriculaire Utero et l'électroporation d'embryons de rat E16

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: En injection intraventriculaire Utero et l'électroporation d'embryons de rat E16

In utero in vivo par injection et l'électroporation du néocortex du rat embryonnaire fournit un outil puissant pour la manipulation des progéniteurs individuelles le long des parois du ventricule latéral. Cette technique est maintenant largement utilisée pour étudier les processus impliqués dans corticogenesis par une sur-exprimant ou faire tomber les gènes et l'observation des effets sur la prolifération cellulaire, la migration et la différenciation. En comparaison avec les stratégies KO traditionnelle, in utero électroporation fournit un moyen rapide de manipuler une population de cellules pendant une fenêtre temporelle spécifique. Dans ce protocole vidéo, nous exposons la méthodologie expérimentale pour la préparation de rats pour la chirurgie, ce qui expose les cornes utérines travers laporatomy, l'injection d'ADN dans les ventricules latéraux de l'embryon en développement, électroporation d'ADN dans le progéniteurs tapissant la paroi latérale, et le soin des animaux d'après- chirurgie. Notre laboratoire utilise ce protocole pour les chirurgies sur des rats E15-E21, cependant il est plus couramment réalisée à E16 comme le montre cette vidéo.

Disclosures: En injection intraventriculaire Utero et l'électroporation d'embryons de rat E16

Ask the Author: En injection intraventriculaire Utero et l'électroporation d'embryons de rat E16

7 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

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

Try different methods of purifying the DNA.

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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?

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Posted by: Chris CunninghamOctober 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.

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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.

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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

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

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

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

what are the medicine for motor neuron disease?

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Posted by: saraSeptember 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?

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Posted by: Julie CardinNovember 10, 2009, 4:19 PM


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

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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!

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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!

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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

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Posted by: Shelley RussekNovember 30, 2011, 11:22 PM

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

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

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

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

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

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

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