Name: a method for labeling vasculature in embryonic mice
Uploaded: 2011-10-07T13:00:00
Description: Watch this Scientific Journal Video about A Method for Labeling Vasculature in Embryonic Mice at JoVE.com

这项工作是支持授权号码R01AI055001和R01AI082127从NIAID的自然资源管理和SREB论文奖学金奖JLB。

1。荧光标记的葡聚糖和金思朗我isolectin乙面部静脉注射标签胚胎血管<ol><li>准备FITC -葡聚糖（50ug/mL）在磷酸盐缓冲液（PBS）或金思朗1 - isolectin乙4（20ug/200uL）在1.5ml的Eppendorf管中，并在PBS温暖至37 ° C 。股票1.25MM快速绿/ PBS 100uL FITC -葡聚糖溶液（总体积毫升）和股票1.25MM快速180uL绿/ PBS金思朗1 - isolectin乙4（总体积200uL），这样的解决方案是明显的蓝色。 </li><li>解剖E14.5 - E18.5胚胎...

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	<li>Cuddihy, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VEGF-mediated+cross-talk+within+the+neonatal+murine+thymus.">VEGF-mediated cross-talk within the neonatal murine thymus.</a> Blood. 113, 2723-2731 (2009).</li><li>Muller, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+targeting+of+VEGF-A+in+thymus+epithelium+disrupts+thymus+blood+vessel+architecture.">Gene targeting of VEGF-A in thymus epithelium disrupts thymus blood vessel architecture.</a> Proc. Natl. Acad. Sci. U. S. A. 102, 10587-10592 (2005).</li><li>Muller, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+crest+origin+of+perivascular+mesenchyme+in+the+adult+thymus.">Neural crest origin of perivascular mesenchyme in the adult thymus.</a> J. Immunol. 180, 5344-5351 (2008).</li><li>Foster, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+neural+crest-derived+cells+in+the+embryonic+and+adult+thymus.">Contribution of neural crest-derived cells in the embryonic and adult thymus.</a> J. Immunol. 180, 3183-3189 (2008).</li><li>Liu, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coordination+between+CCR7-+and+CCR9-mediated+chemokine+signals+in+prevascular+fetal+thymus+colonization.">Coordination between CCR7- and CCR9-mediated chemokine signals in prevascular fetal thymus colonization.</a> Blood. 108, 2531-2539 (2006).</li><li>Lavine, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibroblast+growth+factor+signals+regulate+a+wave+of+Hedgehog+activation+that+is+essential+for+coronary+vascular+development.">Fibroblast growth factor signals regulate a wave of Hedgehog activation that is essential for coronary vascular development.</a> Genes Dev. 20, 1651-1666 (2006).</li><li>Lavine, K. J., Kovacs, A., Ornitz, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hedgehog+signaling+is+critical+for+maintenance+of+the+adult+coronary+vasculature+in+mice.">Hedgehog signaling is critical for maintenance of the adult coronary vasculature in mice.</a> J. Clin Invest. 118, 2404-2414 (2008).</li><li>Mukouyama, Y. S., Gerber, H. P., Ferrara, N., Gu, C., Anderson, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+nerve-derived+VEGF+promotes+arterial+differentiation+via+neuropilin+1-mediated+positive+feedback.">Peripheral nerve-derived VEGF promotes arterial differentiation via neuropilin 1-mediated positive feedback.</a> Development. 132, 941-952 (2005).</li><li>Mukouyama, Y. S., Shin, D., Britsch, S., Taniguchi, M., Anderson, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+nerves+determine+the+pattern+of+arterial+differentiation+and+blood+vessel+branching+in+the+skin.">Sensory nerves determine the pattern of arterial differentiation and blood vessel branching in the skin.</a> Cell. 109, 693-705 (2002).</li><li>Murphy, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+Notch4+signaling+induces+hallmarks+of+brain+arteriovenous+malformations+in+mice.">Endothelial Notch4 signaling induces hallmarks of brain arteriovenous malformations in mice.</a> Proc. Natl. Acad. Sci. U. S. A. 105, 10901-10906 (2008).</li></ol>

全安装和PECAM - 1（CD31）染色部分标签在胚胎小鼠的血管的常规方法。这些方法需要使用的直接和/或间接免疫荧光和洗涤剂通透小鼠组织。这被证明是一个相当及时处理。在这里，我们有FITC -葡聚糖或isolectin B级4面部静脉注射直接标记的胚胎血管，从而消除了抗体标记的步骤要求。此外，这种方法使血管功能状态的评估，将通过这种方法，而不是由传统抗体染色方法显示“漏水”的表型。...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th></tr><tr><td> FITC -葡聚糖</td><td>西格玛</td><td> FD150S - 1G </td></tr><tr><td>荧光标记金思朗1 - 4 isolectin乙</td><td>媒介实验室</td><td> FL - 1201 </td></tr><tr><td>快绿</td><td> MP生物医学</td><td> 195178 </td></tr><tr><td>煤灰</td><td> Fluka公司</td><td> 76240 </td></tr><tr><td>胎牛血清</td><td>亚特兰大生物</td><td> S11550 </td></tr><tr><td>最佳切削温度化合物（华侨城</td><td> VWR </td><td> 25608-930 </td></tr><tr><td>丙酮</td><td> JT贝克</td><td> 9006-33 </td></tr><tr><td>驴血清</td><td>杰克逊</td><td> 017-000-121 </td></tr><tr><td>大鼠抗小鼠CD31， </td><td> BD Pharmingen公司</td><td> 558736 </td></tr><tr><td>羊抗鼠PDGFR -β </td><td> R＆D系统</td><td> AF1042 </td></tr><tr><td>驴抗大鼠CD31的Alexa的647（Invitrogen公司） </td><td> Biolegend </td><td> 102516 </td></tr><tr><td>驴抗山羊Alexa的594（Invitrogen公司） </td><td> Invitrogen公司</td><td> A11058 </td></tr><tr><td> TRITON X -100 </td><td> Sigma - Aldrich公司</td><td> X - 100 </td></tr><tr><td>低熔点琼脂糖/ PBS </td><td> Sigma - Aldrich公司</td><td> A9414 - 25G </td></tr><tr><td>甲醇</td><td> Fisher Scientific则</td><td> A413 - 4 </td></tr><tr><td>苯甲醇</td><td>阿库罗斯福冈科学</td><td> 148390010 </td></tr><tr><td>苯甲酸苄酯</td><td>阿库罗斯福冈科学</td><td> 105860010 </td></tr><tr><td>抑郁症的幻灯片</td><td> Fisher Scientific则</td><td> S175201 </td></tr><tr><td> Fluorogel </td><td>电子显微镜科学</td><td> 17985-10 </td></tr><tr><td>玻璃盖（22X22）-1.5 </td><td> Thermo Scientific的</td><td> 152222 </td></tr><tr><td>蔡司LSM 510 META共聚焦显微镜</td><td>蔡司</td><td></td></tr><tr><td>微解剖钳</td><td> Roboz </td><td> RS - 5135 </td></tr><tr><td>封口膜号OM992 </td><td> Fisher Scientific则</td><td> 13-374-16 </td></tr><tr><td> 12和24孔板</td><td>长荣科学</td><td> 222 - 8044 - 01F </td></tr><tr><td> Superfrost / PLUS显微镜载玻片</td><td> Fisher Scientific则</td><td> 12-550-15 </td></tr><tr><td> 4ML明确小瓶</td><td>国家科学技术进步奖</td><td> B7800 - 2 </td></tr></tbody></table>

a method for labeling vasculature in embryonic mice

功能的血管网络的建立是一个器官形成的重要组成部分，是最佳的器官功能要求。例如，在胸腺适当的血管形成和图案是胸腺细胞进入到器官和成熟的T细胞向周边出口所必需的。在胸腺血管的空间安排，是依赖于从局部微环境的信号，即胸腺上皮细胞（TEC）。最近的一些报告表明，这些信号结果在胸腺血管缺陷1,2中断。以前的研究已经描述技术用于标签的新生儿和成人的胸腺血管 1,2 。我们在这里展示一个标签血管在胚胎胸腺技术。这种方法结合使用FITC -葡聚糖或加纳（Bandeiraea）Simplicifolia凝集素我（1 - GSL isolectin乙4）面部静脉注射和CD31抗体染色，以确定胸腺的血管结构和PDGFR -β标签胸腺血管周围间质 3-5 。还提供了使用冰冻切片或vibratome部分选项。该协议可用于识别胸腺血管的缺陷，这是定义在胸腺血管形成的TEC -派生分子的角色的关键。作为方法的标签，整个血管，它也可以被用来分析多个器官和整个组织，包括皮肤和心脏 6-10胚胎的血管网络。

Watch this Scientific Journal Video about A Method for Labeling Vasculature in Embryonic Mice at JoVE.com

A Method for Labeling Vasculature in Embryonic Mice

本文介绍了标签胚胎皮肤和胸腺血管的方法。

标签胚胎小鼠血管的方法

The establishment of a functional blood vessel network is an essential part of organogenesis, and is required for optimal organ function. For example, in ...

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