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Utero Intraventricular और E16 चूहा भ्रूण के इंजेक्शन electroporation में

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

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Cite this Article: Utero Intraventricular और E16 चूहा भ्रूण के इंजेक्शन electroporation में

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: Utero Intraventricular और E16 चूहा भ्रूण के इंजेक्शन electroporation में

-Utero में vivo और भ्रूण चूहे neocortex के इंजेक्शन electroporation व्यक्तिगत पार्श्व वेंट्रिकल की दीवारों अस्तर progenitors के हेरफेर के लिए एक शक्तिशाली उपकरण प्रदान करता है. इस तकनीक को अब व्यापक रूप से है पर व्यक्त या नीचे जीन दस्तक और सेलुलर प्रसार, प्रवास, और भेदभाव पर प्रभाव देख द्वारा corticogenesis में शामिल प्रक्रियाओं का अध्ययन किया है. पारंपरिक पीटा रणनीतियों की तुलना में utero electroporation एक तेजी से एक विशिष्ट अस्थायी खिड़की के दौरान कक्षों की आबादी में हेरफेर करने का साधन प्रदान करता है. इस वीडियो प्रोटोकॉल में, हम शल्य चिकित्सा के लिए तैयारी चूहों, laporatomy के माध्यम से गर्भाशय सींग उजागर, विकासशील भ्रूण के पार्श्व ventricles में डीएनए इंजेक्शन, पार्श्व दीवार अस्तर progenitors में डीएनए electroporating, और जानवरों की देखभाल करने के लिए प्रयोगात्मक पद्धति रूपरेखा पोस्ट - सर्जरी. हमारी प्रयोगशाला E15-E21 चूहों पर सर्जरी के लिए इस प्रोटोकॉल का उपयोग करता है, लेकिन यह सबसे अधिक E16 पर किया जाता है के रूप में इस वीडियो में दिखाया गया है.

Disclosures: Utero Intraventricular और E16 चूहा भ्रूण के इंजेक्शन electroporation में

Ask the Author: Utero Intraventricular और E16 चूहा भ्रूण के इंजेक्शन electroporation में

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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