E16大鼠胚胎在子宫内侧脑室注射和电穿孔 (Translated to Chinese)

Cite this Article: E16大鼠胚胎在子宫内侧脑室注射和电穿孔

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: E16大鼠胚胎在子宫内侧脑室注射和电穿孔

在子宫内，在体内注射和胚胎大鼠大脑皮层电提供了一个强大的工具操纵个别祖衬侧脑室壁。这项技术现在被广泛用于研究过度表达或撞倒基因，并观察细胞的增殖，迁移和分化的影响，在corticogenesis参与的过程。在比较传统的淘汰赛战略，在子宫内电提供了一个快速的手段操纵人口的细胞，在特定的时间窗口。在此视频协议中，我们概括为准备手术的老鼠，通过laporatomy暴露子宫角注射到发育中的胚胎侧脑室的DNA，电穿孔进入的衬里，侧壁的祖细胞的DNA，和爱护动物的实验方法后手术。我们的实验室使用这个协议对E15 - E21大鼠手术，但它是最常用在E16在这个视频所示。

Disclosures: E16大鼠胚胎在子宫内侧脑室注射和电穿孔

Ask the Author: E16大鼠胚胎在子宫内侧脑室注射和电穿孔

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

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

Try different methods of purifying the DNA.

1.1

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?

1.1.1

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.

1.2

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.

1.2.1

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

1.3

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

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

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

what are the medicine for motor neuron disease?

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?

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

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

4.1

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!

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!

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

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

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

7.1

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!

7.1.1

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!

7.1.2

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

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Date Published	7/19/2007
JoVE Section	General
JoVE Issue	6
Comments	7 Comments
View Count	Loading... (Details…)
DOI	doi:10.3791/236

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E16大鼠胚胎在子宫内侧脑室注射和电穿孔

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