<p class="jove_step">Dissection of reproductive organs from a female mouse</p>
<ol>
<li>Pregnant animals are euthanized by CO<sub>2</sub> asphyxiation using compressed gas. </li>
<li>Lay the animal supine on absorbent pad and soak it in 70% ethanol to reduce the risk of contaminating the dissection with mouse hair. </li>
<li>Pinch the skin and make a small lateral incision at the midline with regular surgical scissors. Hold the skin firmly above and below the incision and pull the skin apart toward the head and tail to expose the abdomen. </li>...

<ol>
	<li>Nagy, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>Manipulating the mouse embryo: A laboratory manual third edition</em>. , (2003).</li></ol>

<p class="jove_content">Postimplantation in the mouse occurs between 4.5 dpc and birth. This dissection method can be applied, essentially as described, to isolate earlier or later stage embryos at 5.5-8.5 dpc.</p>

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Micro Dissecting Forceps serrated, straight</td>
<td>Tool</td>
<td>Roboz</td>
<td>RS-5130 </td>
<td>2 pairs</td>
</tr>
<tr>
<td>Micro Dissecting Tweezers Pattern #55 INOX</td>
<td>Tool</td>
<td>Roboz</td>
<td>RS-5010 </td>
<td>2 pairs</td>
</tr>
<tr>
<td>Dulbecco's Phosphate Buffered Saline (1X) </td>
<td>Reagent</td>
<td>Invitrogen</td>
<td>14190 </td>
<td>&nbsp;</td>
<tr>
<td>Fisherbrand Sterile Polysytrene Petri Dish</td>
<td>Tool</td>
<td>Fisher Scientific</td>
<td>08-757-12 </td>
<td>&nbsp;</td>
<tr>
<td>4% Paraformaldehyde in PBS</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Straight Sharp-Pointed Dissecting Scissors</td>
<td>Tool</td>
<td>Fisher Scientific</td>
<td>08-940 </td>
<td>&nbsp;</td>
<tr>
<td>Pregnant female mice </td>
<td>Animal</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Dissecting Microscope</td>
<td>Microscope</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
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dissection of 6.5 dpc mouse embryos

<p class="jove_content">Analysis of gene expression patterns during early stages of mammalian embryonic development can provide important clues about gene function, cell-cell interaction and signaling mechanisms that guide embryonic patterning. However, dissection of the mouse embryo from the decidua shortly after implantation can be a challenging procedure, and detailed step-by-step documentation of this process is lacking.</p>

<p class="jove_content">Here we demonstrate how post-implantation (6.5 dpc) embryos are isolated by first dissecting the uterus of a pregnant mouse (detection of the vaginal plug was designated day 0.5 poist coitum) and subsequently dissecting the embryo from maternal decidua.  The dissection of Reichert's membrane is described as well as the removal of the ectoplacental cone.</p>

Watch this Scientific Journal Video about Dissection of 6.5 dpc Mouse Embryos at JoVE.com

Dissection of 6.5 dpc Mouse Embryos

<p class="jove_content">Isolation of postimplantation-stage embryos allows one to study gene patterning and analyze cell-lineage decision making processes during embryonic development, but proper dissection of the early embryo can be challenging. This protocol describes a method for isolating early primitive-streak-stage embryos (~6.5 days post coitum [dpc]).</p>

Analysis of gene expression patterns during early stages of mammalian embryonic development can provide important clues about gene function, cell-cell ...

Research

JoVE Journal

Biology

37.9K Görüntüleme.  Harvard Medical School. Postimplantation-dönem embriyoların İzolasyon bir gen desenlendirme hücre soy karar verme süreçlerini embriyonik gelişim sırasında çalışma ve analiz etmek için izin verir, fakat erken embriyo uygun diseksiyonu zor olabilir. Bu protokol erken ilkel bir çizgi-dönem embriyoların izole etmek için bir yöntem açıklanır (~ 6.5 gün coitum [DPC).