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في الرحم وحقن داخل البطيني Electroporation الجنين الجرذ E16

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

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Cite this Article: في الرحم وحقن داخل البطيني Electroporation الجنين الجرذ 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: في الرحم وحقن داخل البطيني Electroporation الجنين الجرذ E16

داخل الرحم في الجسم الحي ، وحقن electroporation من القشرة المخية الحديثة الفئران الجنينية يوفر أداة قوية للتلاعب من أسلاف الفردية التي تبطن جدران البطين الجانبي. الآن هذه التقنية المستخدمة على نطاق واسع لدراسة العمليات التي ينطوي عليها corticogenesis بالإعراب عن الإفراط أو هدمت الجينات ومراقبة الآثار المترتبة على انتشار الخليوي ، والهجرة ، والتمايز. في مقارنة لاستراتيجيات خروج المغلوب التقليدية ، في الرحم ، electroporation يوفر وسيلة سريعة لمعالجة مجموعة من الخلايا خلال نافذة زمنية محددة. في هذا البروتوكول الفيديو ، ونحن الخطوط العريضة لمنهجية تجريبية لاعداد الفئران للجراحة ، ويعرض من خلال أبواق الرحم laporatomy ، حقن الحمض النووي في البطينات الجانبية للجنين النامي ، electroporating أسلاف الحمض النووي في بطانة الجدار الجانبي ، ورعاية للحيوانات في مرحلة ما بعد الجراحة. مختبرنا يستخدم هذا البروتوكول لعملية جراحية في E15 - E21 الفئران ، ولكن هو الأكثر شيوعا تنفيذ ذلك في E16 كما هو مبين في هذا الفيديو.

Disclosures: في الرحم وحقن داخل البطيني Electroporation الجنين الجرذ E16

Ask the Author: في الرحم وحقن داخل البطيني Electroporation الجنين الجرذ 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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