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בשנת הזרקה תוך חדרי ברחם ו electroporation של E16 עוברים עכברוש

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

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Cite this Article: בשנת הזרקה תוך חדרי ברחם ו electroporation של E16 עוברים עכברוש

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: בשנת הזרקה תוך חדרי ברחם ו electroporation של E16 עוברים עכברוש

ברחם-in-vivo הזרקת electroporation של הניאוקורטקס עכברוש עובריים מספק כלי רב עוצמה עבור מניפולציה של אבות בודדים לאורך הקירות של החדר לרוחב. טכניקה זו היא כעת בשימוש נרחב כדי לחקור את התהליכים המעורבים corticogenesis על ידי יתר להביע או דריסה גנים התבוננות על תופעות התפשטות ההגירה הסלולר, והבחנה. בהשוואה אסטרטגיות בנוקאאוט המסורתית, ב-ברחם electroporation מספק אמצעי מהיר כדי לתפעל אוכלוסייה של תאים במהלך חלון הזמני ספציפיים. בפרוטוקול זה וידאו, אנחנו המתאר את המתודולוגיה ניסיוני להכנת חולדות לניתוח, לחשוף את הקרניים דרך הרחם laporatomy, הזרקת DNA לתוך החדרים רוחבי של העובר המתפתח, electroporating דנ"א לתוך אבות המצפים את הקיר לרוחב, וטיפול בבעלי חיים שלאחר הניתוח. המעבדה שלנו משתמשת בפרוטוקול זה על ניתוחים על E15-E21 חולדות, אולם זה מבוצע לרוב על E16 כפי שמוצג בסרטון זה.

Disclosures: בשנת הזרקה תוך חדרי ברחם ו electroporation של E16 עוברים עכברוש

Ask the Author: בשנת הזרקה תוך חדרי ברחם ו electroporation של E16 עוברים עכברוש

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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