Name: placing growth factor-coated beads on early stage chicken embryos
Uploaded: 2007-10-01T00:00:00
Description: Watch this Scientific Journal Video about Placing Growth Factor-Coated Beads on Early Stage Chicken Embryos at JoVE.com

<p class="jove_step"> 1。取出鸡蛋从孵化器</p><p class="jove_content">在E2 - 2.5（〜圣17 HH），去掉鸡蛋和相应准备。他们可工作立即或几个小时的孵化器。鸡蛋可以安全地放置在室温下了一个多小时。鸡蛋可以留出操作的持续时间。有时，这些都需要一个小时或更长时间。 </p><p class="jove_content"> <em>注</em> ：他们在室温下坐的时间越长，不太可能，他们要生存。 </p><p class="jove_step"> 2。印度墨水的制备</p><ol><li>准备新鲜的解决?...

<ol>
	<li>Korn, M. J., Cramer, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Windowing+Chicken+Eggs+for+Developmental+Studies.">Windowing Chicken Eggs for Developmental Studies.</a> <em style='font-style: italic'>Jounal of Visualized Experiments</em>. <b>8</b>, (2007).</li><li>Crossley, P. H., Martinez, S., Ohkubo, Y., Rubenstein, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coordinate+expression+of+Fgf8,+Otx2,+Bmp4,+and+Shh+in+the+rostral+prosencephalon+during+development+of+the+telencephalic+and+optic+vesicles.">Coordinate expression of Fgf8, Otx2, Bmp4, and Shh in the rostral prosencephalon during development of the telencephalic and optic vesicles.</a> <em style='font-style: italic'>Neuroscience</em>. <b>108</b>, 183-206 (2001).</li><li>Alexandre, P., Bachy, I., Marcou, M., Wassef, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Positive+and+negative+regulations+by+FGF8+contribute+to+midbrain+roof+plate+developmental+plasticity.">Positive and negative regulations by FGF8 contribute to midbrain roof plate developmental plasticity.</a> <em style='font-style: italic'>Development </em>. <b>133</b>, 2905-2913 (2006).</li></ol>

<p>根据您要使用的蛋白质，有不同的协议，关于如何准备珠。在这些实验中，我们使用肝素丙烯酸珠，但也可用于阿菲 - 凝胶珠。我们发现，肝素丙烯酸珠更容易操纵在水环境中的卵白蛋白。珠蛋白质在5升浸泡在4 ° C过夜控制珠被放置在无菌PBS，直到放置时间。在使用OVO准备的优点是，胚胎仍然活着，和自由发展<sup>通常</sup>为1。虽然保存在其中的一个外植体的环境，可用于特定变量控制的，活的胚胎允许<sup>的<...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Eggs</td>
<td>Animal</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Fertilized, E2-2.5 (~ St. 17 HH)
</td>
</tr>
<tr>
<td>Incubator</td>
<td>Profi-I</td>
<td>Lyon</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>India Ink</td>
<td>Tool</td>
<td>Higgins</td>
<td>&nbsp;</td>
<td>Prepare a 4% solution in sterile PBS.</td>
</tr>
<tr>
<td>Sterile PBS</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Alternatively, an antibody cocktail can be used</td>
</tr>
<tr>
<td>1ml syringe</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>27G 1/2 needles</td>
<td>Tool</td>
<td>BD</td>
<td>305109</td>
<td>Angled at 90 degrees</td>
</tr>
<tr>
<td>Forceps</td>
<td>Tool</td>
<td>World Precision Instruments</td>
<td>&nbsp;</td>
<td>size 55</td>
</tr>
<tr>
<td>Hemostats</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Dissection scissors</td>
<td>Tool</td>
<td>World Precision Instruments</td>
<td>500086</td>
<td>vannas, 8.5cm</td>
</tr>
<tr>
<td>Parafilm tape</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>to seal eggs</td>
</tr>
<tr>
<td>Tungsten Probe</td>
<td>Tool</td>
<td>Electron Microscopy Sciences</td>
<td>62091</td>
<td>Tool #24.  Handle optional. </td>
</tr>		</tbody>
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placing growth factor-coated beads on early stage chicken embryos

<p class="jove_content">神经管表示在特定的时空模式，在开发过程中的多种蛋白质。这些蛋白质已被证明是对细胞命运的决心，细胞迁移，并形成神经回路至关重要。神经元的诱导和图案涉及骨形态发生蛋白（BMP），刺猬（SHH），成纤维细胞生长因子（FGF），等等。 FGF8的表达模式，尤其是在邻近地区表达BMP4和SHH。这种表达模式与发展的互动，以促进在端脑的图案一致。 </p><p class="jove_content">在这里，我们提供的视觉示范在其中OVO准备一个可以用来测试在形成前脑Fgfs的影响的方法。珠涂有蛋白质和发展中的神经管提供持续的曝光。因为程序使用小，小心地放在珠，这是微创，并允许被放置在一个单一的神经管几个珠子。此外，该方法可以使胚胎可以在一个比较成熟的的阶段分析，以检测在解剖学和神经图案的变化，持续发展。这一简单实用的协议允许实时成像。它提供了一种手段，使内源性蛋白水平的空间和时间有限的修改。 </p>

Watch this Scientific Journal Video about Placing Growth Factor-Coated Beads on Early Stage Chicken Embryos at JoVE.com

Placing Growth Factor-Coated Beads on Early Stage Chicken Embryos

<p class="jove_content">多种生长因子和蛋白质相互作用，从而诱导细胞在发展过程中不同的细胞命运。在这里，我们展示了一个在OVO准备使用，以解决发展中的不同的蛋白质之间可能的相互作用，由珠E2.5鸡胚。</p>

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