Name: using a whole-mount immunohistochemical method to study the innervation of the biliary tract in suncus murinus
Uploaded: 2017-06-15T12:30:00
Description: Watch this Scientific Journal Video about Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus at JoVE.com

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所有实验程序均由东京都大学体育动物护理与使用委员会（IACUC）批准。根据“实验动物护理和使用 指南”和 “加拿大动物护理委员会实验动物护理和使用指南”，将动物饲养和处理。我们在日本东京都大学前沿健康科学系功能形态学实验室养殖和维护动物。 动物护理和住房<ol><li>从封闭的繁殖群中获得7只小鼠 （3只雌性和4只雄性，重70-90克）。...

<ol>
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Q., 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=Surgical+anatomy+of+the+innervation+of+the+pylorus+in+humans+and+Suncus+murinus,+in+relation+to+the+surgical+technique+for+pylorus-preserving+pancreatoduodenectomy.">Surgical anatomy of the innervation of the pylorus in humans and Suncus murinus, in relation to the surgical technique for pylorus-preserving pancreatoduodenectomy.</a> World J. Gastroenterol. 12 (14), 2209-2216 (2006).</li><li>Yi, S. 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这项工作描述了使用抗神经丝蛋白的抗体对肝外胆道神经支配的可视化的实验程序。该协议根据Kuratani和Tanaka 1，2，3描述的方案进行了改编。 对于这个实验，计划每个实验的时间表，因为整体染色方法持续2-3周。包含组织的容器必须始终在营养器上旋转，除非在冻融过程中。特别是在与一级和二级抗体一起孵育的同时，将样品置于冷藏室中，同时在养分器?...

房子麝香rew，，，属于食虫纲和家Sor科。这种小型哺乳动物分布在整个东南亚，栖息在靠近人类栖息地或牛栏的房屋和草地上。与其他实验室动物（如小鼠，大鼠和兔5）相比，该物种具有与人类相似的一般形态特征。以前，使用具有S. murinus神经丝蛋白（NFP）的外周神经元标记的全蛋白免疫染色来标记胰腺6 ，主要十二指肠乳头7 ，幽门8 ，胆囊5和肝外胆道9中的外周神经。 NFP是成熟神经元细胞骨架的一部分的大分子复合物。神经丝相关蛋白介导NFP和Zygosome之间的相互作用。酶功能和接头蛋白的结构均受NFP 10调控。 NFP复合物由三种多肽组成：NF-L（70kDa），NF-M（150-160kDa）和NF-H（200kDa）。 NFP可以在中枢神经系统和外周神经系统的神经元轴突中发现。已经显示抗人NFP抗体标记中枢和周围神经系统的轴突。解剖和电生理调查表明，括约肌，胆囊和近端胃肠道连接11,12,13,14。然而，他们的神经元连接的形态特征尚未被定义。 这项工作表明使用全套免疫组织化学染色方法来标记耻骨球菌的神经支配使用NFP抗体。

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">orthoperiodic acid</td>
			<td style="width:71px;">Wako</td>
			<td style="width:119px;">162-00732</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">papain</td>
			<td style="width:71px;">Wako</td>
			<td style="width:119px;">164-00172</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">bovine serum albumin (BSA)</td>
			<td style="width:71px;">Wako</td>
			<td style="width:119px;">010-15131</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">sodium azide</td>
			<td style="width:71px;">Nacalai Tesque</td>
			<td style="width:119px;">312-33</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">neurofilament protein (NFP) antibody</td>
			<td style="width:71px;">Dako</td>
			<td style="width:119px;">M0762, lot 089, clone: 2F11</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">peroxidase-conjugated affinity-purified sheep anti-mouse IgG-HRP</td>
			<td style="width:71px;">MBL</td>
			<td style="width:119px;">Code 330</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">3,3&#39;-diaminobenzidine tetrahydrochloride (DAB)</td>
			<td style="width:71px;">Wako</td>
			<td style="width:119px;">349-00903</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">imidazole</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">I-0125</td>
			<td></td>
		</tr>
	</tbody>
</table>

using a whole-mount immunohistochemical method to study the innervation of the biliary tract in suncus murinus

本文详细描述了全基因免疫组织化学染色方法，使用神经丝蛋白抗体标记阳性丝猴（ S. murinus）胆道神经支配 &#160; ）。首先从S. murinus中切下标本并固定在4％多聚甲醛（PFA）中。其次，进行酶处理和潜在的内源性过氧化物酶失活。然后将标本暴露于一抗，抗神经丝蛋白抗体3-6天。然后与与辣根过氧化物酶缀合的二抗孵育。通过使样品与3,3&#39;-二氨基联苯胺（DAB）底物反应来显示显色反应。该方法可用于分析所有内脏器官的神经支配。此外，该方案还可以适应于测试其他神经元抗体，但优化抗体应先做好。这种方法最初由Kuratani和Tanaka 1，2，3引入。

图 1-4显示了S. murinus肝外胆道中NFP阳性神经纤维的典型结果。抗NFP的抗体可重复标记肝外胆管（ 图1 ），胆囊（ 图2 ），上胆管（ 图3 ）和十二指肠乳头区域（ 图4 ）的整个图像中的神经支配，具有高特异性和最小背景。 <p class="jove_content" fo:keep-to...

Watch this Scientific Journal Video about Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus at JoVE.com

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus

一种全身免疫组织化学方法，可以显示Suncus murinus肝外胆道中的神经丝蛋白表达。在这里介绍。该方案可用于分析小鼠或其他物种中所有内脏器官的神经支配。

使用全身免疫组织化学方法研究胆道的神经支配<em&gt; Suncus murinus</em

This work describes the whole-mount immunohistochemistry staining method in detail, using neurofilament protein antibody to label the innervation of the ...

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JoVE Journal

Biology

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus

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Testing visual sensitivity in any species provides basic information regarding behaviour, evolution, and ecology. However, testing specific features of ...

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Whole mount in situ hybridization is one of the most widely used techniques in developmental biology.  Here, we present a high-resolution double ...

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The three-dimensional folding of chromosomes compartmentalizes the genome and and can bring distant functional elements, such as promoters and enhancers, ...

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Whole mount in situ hybridization is a very informative approach for defining gene expression patterns in embryos. The in situ hybridization procedures ...

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Development of the lower urogenital tract (LUT) is an intricate process. This complexity is evidenced during formation of the prostate from the fetal male ...

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This article focuses on whole-mount in situ hybridization (WISH) of zebrafish embryos. The WISH technology facilitates the assessment of gene expression ...