Name: glomerular outgrowth as an ex vivo assay to analyze pathways involved in parietal epithelial cell activation
Uploaded: 2020-08-19T13:00:00
Description: Watch this Scientific Journal Video about Glomerular Outgrowth as an Ex Vivo Assay to Analyze Pathways Involved in Parietal Epithelial Cell Activation at JoVE.com

这项研究得到了荷兰肾脏基金会（赠款14A3D104）和荷兰科学研究组织（NWO VIDI赠款：016.156.363）的支持。

所有动物实验都是按照拉德布德大学尼梅根大学动物伦理委员会的指导方针进行的。
注：未经治疗的、健康的野生型小鼠（n = 4） 和cd44-/--（n = 4）小鼠在12~16周龄被牺牲。使用雄性小鼠和雌性小鼠。所有小鼠都在C57Bl/6背景。
1. 小鼠肾脏解剖
<ol><li>通过宫颈脱位牺牲健康的WT小鼠或基因改变的小鼠。</li>
	<li>牺牲老鼠后直接解剖整个小鼠肾脏。为?...

<ol>
	<li>Fatima, H., 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=Parietal+epithelial+cell+activation+marker+in+early+recurrence+of+FSGS+in+the+transplant.">Parietal epithelial cell activation marker in early recurrence of FSGS in the transplant.</a> Clinical journal of the American Society of Nephrology: CJASN. 7 (11), 1852-1858 (2012).</li><li>Dijkman, H. B., 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=Proliferating+cells+in+HIV+and+pamidronate-associated+collapsing+focal+segmental+glomerulosclerosis+are+parietal+epithelial+cells.">Proliferating cells in HIV and pamidronate-associated collapsing focal segmental glomerulosclerosis are parietal epithelial cells.</a> Kidney International. 70 (2), 338-344 (2006).</li><li>Kuppe, C., 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=Common+histological+patterns+in+glomerular+epithelial+cells+in+secondary+focal+segmental+glomerulosclerosis.">Common histological patterns in glomerular epithelial cells in secondary focal segmental glomerulosclerosis.</a> Kidney International. 88 (5), 990-998 (2015).</li><li>Dijkman, H., Smeets, B., van der Laak, J., Steenbergen, E., Wetzels, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+parietal+epithelial+cell+is+crucially+involved+in+human+idiopathic+focal+segmental+glomerulosclerosis.">The parietal epithelial cell is crucially involved in human idiopathic focal segmental glomerulosclerosis.</a> Kidney International. 68 (4), 1562-1572 (2005).</li><li>Smeets, B., 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=Parietal+epithelial+cells+participate+in+the+formation+of+sclerotic+lesions+in+focal+segmental+glomerulosclerosis.">Parietal epithelial cells participate in the formation of sclerotic lesions in focal segmental glomerulosclerosis.</a> Journal of the American Society of Nephrology: JASN. 22 (7), 1262-1274 (2011).</li><li>Smeets, B., 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=Tracing+the+origin+of+glomerular+extracapillary+lesions+from+parietal+epithelial+cells.">Tracing the origin of glomerular extracapillary lesions from parietal epithelial cells.</a> Journal of the American Society of Nephrology: JASN. 20 (12), 2604-2615 (2009).</li><li>Smeets, B., Moeller, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parietal+epithelial+cells+and+podocytes+in+glomerular+diseases.">Parietal epithelial cells and podocytes in glomerular diseases.</a> Seminars in Nephrology. 32 (4), 357-367 (2012).</li><li>Ryu, M., 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=Plasma+leakage+through+glomerular+basement+membrane+ruptures+triggers+the+proliferation+of+parietal+epithelial+cells+and+crescent+formation+in+non-inflammatory+glomerular+injury.">Plasma leakage through glomerular basement membrane ruptures triggers the proliferation of parietal epithelial cells and crescent formation in non-inflammatory glomerular injury.</a> The Journal of Pathology. 228 (4), 482-494 (2012).</li><li>Eymael, J., Smeets, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Origin+and+fate+of+the+regenerating+cells+of+the+kidney.">Origin and fate of the regenerating cells of the kidney.</a> European Journal of Pharmacology. 790, 62-73 (2016).</li><li>Smeets, B., 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=Renal+progenitor+cells+contribute+to+hyperplastic+lesions+of+podocytopathies+and+crescentic+glomerulonephritis.">Renal progenitor cells contribute to hyperplastic lesions of podocytopathies and crescentic glomerulonephritis.</a> Journal of the American Society of Nephrology: JASN. 20 (12), 2593-2603 (2009).</li><li>Eymael, J., 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=CD44+is+required+for+the+pathogenesis+of+experimental+crescentic+glomerulonephritis+and+collapsing+focal+segmental+glomerulosclerosis.">CD44 is required for the pathogenesis of experimental crescentic glomerulonephritis and collapsing focal segmental glomerulosclerosis.</a> Kidney International. 93 (3), 626-642 (2018).</li><li>Roeder, S. S., 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=Activated+ERK1/2+increases+CD44+in+glomerular+parietal+epithelial+cells+leading+to+matrix+expansion.">Activated ERK1/2 increases CD44 in glomerular parietal epithelial cells leading to matrix expansion.</a> Kidney International. 91 (4), 896-913 (2017).</li><li>Appel, D., 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=Recruitment+of+podocytes+from+glomerular+parietal+epithelial+cells.">Recruitment of podocytes from glomerular parietal epithelial cells.</a> Journal of the American Society of Nephrology: JASN. 20 (2), 333-343 (2009).</li><li>Yaoita, E., 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=Visceral+epithelial+cells+in+rat+glomerular+cell+culture.">Visceral epithelial cells in rat glomerular cell culture.</a> European Journal of Cell Biology. 67 (2), 136-144 (1995).</li><li>Kuppe, C., 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=Investigations+of+Glucocorticoid+Action+in+GN.">Investigations of Glucocorticoid Action in GN.</a> Journal of the American Society of Nephrology: JASN. 28 (5), 1408-1420 (2017).</li><li>Ohse, T., 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=Establishment+of+conditionally+immortalized+mouse+glomerular+parietal+epithelial+cells+in+culture.">Establishment of conditionally immortalized mouse glomerular parietal epithelial cells in culture.</a> Journal of the American Society of Nephrology: JASN. 19 (10), 1879-1890 (2008).</li></ol>

使用本文中描述的协议，可以使用单封装的球状细胞来评估单体上皮细胞增殖，这是单体上皮细胞激活的结果。这个外活体模型将使我们能够详细研究分子通路，其中涉及表皮上皮细胞活化。所述方法依靠肾脏解剖和分离的简单概念来分离和培养封装的球蛋白，并比较不同实验条件下皮皮上皮细胞的增殖和/或迁移。在培养中6天后可以分析的结果是，例如，生长的表面积或直径，或单个帽状球蛋?...

肾球疾病是肾病的重要群体，是最终期肾病（ESRD）的一个根本原因。不幸的是，具体的治疗选择是有限的，进展到ESRD是不可避免的。球状疾病由球状损伤的存在定义，可分组在炎症和非炎症性疾病中。虽然最初的侮辱是不同的，最近的研究表明，一个共同的细胞机制导致球状上皮细胞增生，并最终导致所有球状疾病的球状硬化症，无论根本原因1，2，3，4。2,3,41
具体来说，它表明，球状硬化病变主要由活化的表皮上皮细胞5，6,组成。在生理条件下，表皮上皮细胞是平的静态上皮细胞，与鲍曼的球状胶囊一起。然而，任何由于基因突变（如足细胞特异性或线粒体细胞病变）、炎症或超滤（例如，由肾质量降低、高血压、肥胖或糖尿病肌酸引起的肾球菌损伤）都可能引发表皮上皮细胞的激活。活性的表皮上皮细胞增殖并沉积细胞外基质，导致细胞新月或,硬化病变5、7、87的形成。5这些过程的进展导致肾功能丧失9。因此，在炎症和非炎症性球菌疾病1、2、3、4、10中，皮皮上皮细胞活化是球,2,3,菌硬化的发育和进展的关键因素。
分子过程调解表皮上皮细胞活化仍然在很大程度上未知。最近的研究表明，激活的皮皮上皮细胞d novo快递CD44，一种受体，是重要的激活参与细胞增殖和迁移的不同途径。此外，CD44的抑制作用被证明能抑制表皮上皮细胞活化，并减轻炎症和非炎症性球菌病动物模型中新月形成和球状硬化的进展。,12
由于单体上皮细胞活化是发展球状硬化和新月形成的关键技术，抑制这些细胞可以减缓球状疾病的进展。促进单体上皮细胞活化的分子通路的阐释可能导致特定治疗干预措施的发展，减少球状疾病中超塑料和球状硬化病变的形成。
在实验动物模型中，通常很难提供证据，证明基因表达的改变（淘汰模型或转基因小鼠模型）或药物治疗对表皮上皮细胞的直接影响。在传统的敲出小鼠中，观察到的体内变化可以通过皮皮上皮细胞的直接变化来解释。然而，由于基因表达在小鼠中的其他细胞类型中也发生了变化，因此不能排除由其他细胞类型调解的间接效应。由主要活跃在表皮上皮细胞中的启动子驱动的有条件的 cre-lox 小鼠的发展，在某些情况下提供了一个解决方案。然而，有条件的转基因模型是复杂的，虽然有更多的条件线可用，对于许多传统的淘汰或转基因小鼠线，还没有一个有条件的替代品。
为了研究对单体上皮细胞的直接影响，我们组开发了一种前体外测定，使用从小鼠肾脏分离出的单封装球蛋白来测量和分析表皮上皮细胞增殖和迁移。此方法将使我们能够确定单皮上皮细胞特异性效应，并找到负责任的途径，为单皮上皮细胞激活和测试治疗选项，以抑制这种激活。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr >
			<td >24-well cell culture plate</td>
			<td >Corning Costar</td>
			<td ></td>
		</tr>
		<tr >
			<td >anti-CD31</td>
			<td>BD Pharmingen</td>
			<td>Endothelial cell marker (used concentration 1:200)</td>
		</tr>
		<tr >
			<td >chicken-anti-rat Alexa 647</td>
			<td>Thermo Fisher</td>
			<td>&#160;(used concentration 1:200)</td>
		</tr>
		<tr >
			<td >DAPI-Fluoromount G</td>
			<td>Southern Biotech</td>
			<td>Mounting medium containing DAPI</td>
		</tr>
		<tr >
			<td >Digital inverted light microscope</td>
			<td>Westburg, EVOS fl microscope</td>
			<td></td>
		</tr>
		<tr >
			<td >donkey-anti-goat Alexa 568</td>
			<td>Thermo Fisher</td>
			<td>&#160;(used concentration 1:200)</td>
		</tr>
		<tr >
			<td >donkey-anti-rabbit Alexa 568</td>
			<td>Thermo Fisher</td>
			<td>&#160;(used concentration 1:200)</td>
		</tr>
		<tr >
			<td >Dulbecco&#39;s Modified Eagle&#39;s medium&#160;</td>
			<td>Lonza</td>
			<td></td>
		</tr>
		<tr >
			<td >EBM Medium</td>
			<td>Lonza</td>
			<td></td>
		</tr>
		<tr >
			<td >EBM-MV Single Quots kit</td>
			<td>Lonza</td>
			<td>containing hydrocortisone, hEGF, GA-1000, FBS and BBE</td>
		</tr>
		<tr >
			<td >Fetal Bovine Serum</td>
			<td>Lonza</td>
			<td></td>
		</tr>
		<tr >
			<td >Fetal Calf Serum</td>
			<td>Lonza</td>
			<td></td>
		</tr>
		<tr >
			<td >Fluorescent microscope</td>
			<td>Leica Microsystems GmbH</td>
			<td></td>
		</tr>
		<tr >
			<td >goat-anti-synaptopodin</td>
			<td>Santa Cruz</td>
			<td>Podocyte marker (used concentration 1:200)</td>
		</tr>
		<tr >
			<td >Hanks&#39;Balanced Salt Solution</td>
			<td>Gibco</td>
			<td></td>
		</tr>
		<tr >
			<td >ImageJ software</td>
			<td >FIJI 1.51n</td>
			<td></td>
		</tr>
		<tr >
			<td >petri dish</td>
			<td >Sarstedt</td>
			<td>size 100</td>
		</tr>
		<tr >
			<td >rabbit-anti-claudin1</td>
			<td>Abcam</td>
			<td>Parietal epithelial cell marker (used concentration 1:100)</td>
		</tr>
		<tr >
			<td >rabbit-anti-SSeCKS</td>
			<td>Roswell Park Comprehensive Cancer Center,Buffalo, NY, USA</td>
			<td>kindly provided by Dr. E. Gelman, Parietal epithelial cell marker</td>
		</tr>
		<tr >
			<td >rat-anti-CD44</td>
			<td>BD Pharmingen</td>
			<td>Parietal epithelial cell marker (used concentration 1:200)</td>
		</tr>
		<tr >
			<td >scalpel</td>
			<td >Dahlhausen</td>
			<td>size 10</td>
		</tr>
		<tr >
			<td >Sieves&#160;</td>
			<td >Endecotts Ltd</td>
			<td>size 300 &#181;m, 75 &#181;m, 53 &#181;m, steel</td>
		</tr>
		<tr >
			<td >syringe</td>
			<td >BD Plastipak</td>
			<td>size: 20 ml</td>
		</tr>
		<tr >
			<td >Ultra-Low Attachment Microplates&#160;</td>
			<td>Corning Costar</td>
			<td>6-well plates</td>
		</tr>
	</tbody>
</table>

glomerular outgrowth as an ex vivo assay to analyze pathways involved in parietal epithelial cell activation

表皮上皮细胞（PEC）活化是影响球菌硬化发育和进展的关键因素之一。因此，抑制参与表皮上皮细胞活化的通路可能是减轻肾上腺疾病进展的工具。本文介绍了一种培养和分析从小鼠肾脏分离出的封装球状细胞生长的方法。解剖分离的小鼠肾脏后，组织被切碎，通过分离分离球蛋白。采集包皮球蛋白，培养单个球蛋白6天，获得肾上皮细胞的球状生长。在此期间，可以通过确定外显细胞的细胞数量或表面积来分析表皮上皮细胞增殖和迁移。因此，这种测定可以作为一种工具，用于研究转基因或敲敲小鼠基因表达的改变，或培养条件对表皮上皮细胞生长特性和信号的影响。使用这种方法，可以研究涉及表皮上皮细胞活化过程以及因此在球蛋白硬化过程中的重要通路。

图1显示了执行球状生长分析方法的系统图。图 2A-D显示了在使用光显微镜观察到的不同时间点的封装球状球蛋白的球状增长。在从小鼠肾脏分离后，在培养中的第2天、第4天和6天（图2B-D）显示出树。为了验证外生长的细胞是表皮上皮细胞，斩首的球状体也分离和培养了6天，如图<...

Watch this Scientific Journal Video about Glomerular Outgrowth as an Ex Vivo Assay to Analyze Pathways Involved in Parietal Epithelial Cell Activation at JoVE.com

Glomerular Outgrowth as an Ex Vivo Assay to Analyze Pathways Involved in Parietal Epithelial Cell Activation

本文介绍了一种培养和分析从小鼠肾脏分离出的封装球状上皮细胞生长的方法。此方法可用于研究涉及单骨上皮细胞增殖和迁移的途径。

球状外生长作为前维沃分析分析涉及的路径的表皮上皮细胞激活

Parietal epithelial cell (PEC) activation is one of the key factors involved in the development and progression of glomerulosclerosis. Inhibition of ...

表皮上皮细胞的激活是肾脏球状疤痕组织发育的关键因素。使用这种方法，我们可以研究这种激活所涉及的细胞过程，并希望找到治疗方案来减缓甚至阻止疾病进展。我们技术的主要优点是，我们使用直接从孤立的球状细胞生长的原细胞。释放肾脏后，如文本手稿中概述的，用手术钳握住肾脏，并使用另一对钳子拉出肾胶囊。在此之后，将肾脏放入六孔培养板的井中，每个孔包含两毫升的 HBSS，并放在冰上培养板上。首先，将肾脏转移到100毫米的培养皿中，用两个手术刀把它们切成大约一到两毫米的小碎片。用HSS保持碎肾片湿润。接下来，将肾片放在 300 微米金属筛的顶部，并使用 20 毫升注射器的柱塞压压肾脏。在两者之间用HSS反复冲洗筛子，并使用血清移液器收集流经并转移到干净的培养皿中。使用手术刀刮掉筛子底部的一切，并转移到收集的肾脏同质性。用 HBSS 通过 75 和 53 微米筛冲洗肾脏均质。然后，用HSSS洗洗两个筛子，以去除所有较小的结构。通过用DMEM补充20%胎儿小腿血清清洗每个筛子的上表面，收集留在筛子中的肾脏结构和材料，将材料转移到六孔超低附着微孔板的孔中。将超低附件微孔板带到倒置的光学显微镜上，并使用 20 微升移液器采集单封装或斩首的 glomeruli。在移液器尖端中捕获单个球状物后，将新的 DMEM 介质添加到同一移液器尖端中，以达到 20 微升。将用DMEM介质的单球体转移到24井细胞培养板的中心，在37摄氏度下用5%的二氧化碳孵育3小时，使球状体附着在井的中心。孵育后，球状体将附着在井的中心。仔细添加500微升内皮基底介质，辅以生长因子套件和额外的5%FBS和1%青霉素和链霉素到每个井。培养在37摄氏度的单球与5%的二氧化碳6天。经过六天的孵化，使用数字倒置光显微镜拍摄图像并分析球状生长。使用图像分析软件，通过打开一个带比例线的图像文件来确定光洁的表面积。在比例栏上绘制一条直线，然后单击"分析"，然后测量以确定以像素为单位的距离。要确定比例，请单击"分析并设置比例"，然后输入已知距离（以像素为单位）、已知距离和长度单位。完成后单击"确定"。单击"分析和设置测量"。然后，检查"区域"和"显示标签"选项。完成后单击"确定"。接下来，在球状生长周围绘制一个徒手选择。单击"分析"和"度量"以显示结果表，该表包含先前确定比例中增长的表面积。在这项研究中，从小鼠肾脏分离、培养和分析包状球状上皮细胞的遗传性上皮细胞生长。代表性光显微镜图像显示，球状与小鼠肾脏分离后，在培养过程中不同时间点的球状生长。为了验证外生长细胞是表皮上皮细胞，被斩首的球状糖也被分离并培养六天。在这个潜伏期，被斩首的球蛋白没有细胞生长。然后，针对不同的表皮上皮细胞标记、照片位点特定标记以及内皮细胞标记进行免疫荧光染色。结果证实，外生长的细胞，事实上，是表皮上皮细胞。与从野生型小鼠分离出的球状小鼠相比，CD44敲除小鼠的球状小鼠显示，生长的细胞数量减少，球状生长表面积也减少。这表明CD44在表皮上皮细胞活化中具有重要作用。使用这种技术，它也可以研究对药物对单皮上皮细胞活化的影响。这可以通过用您选择的药物治疗孤立的球菌，并分析细胞生长和细胞迁移的差异。

Research

JoVE Journal

Biology

Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells

Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells

前初级人类输卵管上皮细胞的体外培养

Epithelial ovarian cancer is a leading cause of female cancer mortality in the United States. In contrast to other women-specific cancers, like breast and ...

科研

生物学

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

激活细胞凋亡的细胞色素的细胞质显微注射 C</em

Apoptosis, or programmed cell death, is a conserved and highly regulated pathway by which cells die1.  Apoptosis can be triggered when cells encounter a ...

Chromatin Isolation by RNA Purification (ChIRP)

Chromatin Isolation by RNA Purification ChIRP

RNA纯化的染色质隔离（CHIRP）

Long noncoding RNAs are key regulators of chromatin states for important biological processes such as dosage compensation, imprinting, and developmental ...

Rapid Genetic Analysis of Epithelial-Mesenchymal Signaling During Hair Regeneration

在毛发再生的上皮 - 间充质信号的快速遗传分析

Hair follicle morphogenesis, a complex process requiring interaction between epithelia-derived keratinocytes and the underlying mesenchyme, is an ...

An Ex vivo Culture System to Study Thyroid Development

An Ex vivo Culture System to Study Thyroid Development

一个<em&gt;体外</em&gt;企业文化系统来研究甲状腺发展

The thyroid is a bilobated endocrine gland localized at the base of the neck, producing the thyroid hormones T3, T4, and calcitonin. T3 and T4 are ...

Mouse Fetal Whole Intestine Culture System for Ex Vivo Manipulation of Signaling Pathways and Three-dimensional Live Imaging of Villus Development

Mouse Fetal Whole Intestine Culture System for Ex Vivo Manipulation of Signaling Pathways and Three-dimensional Live Imaging of Villus Development

老鼠胎儿整肠文化系统<em&gt;前体内</em&gt;信号通路和绒毛开发的三维实时成像的操作

Most morphogenetic processes in the fetal intestine have been inferred from thin sections of fixed tissues, providing snapshots of changes over ...

Real Time Analysis of Metabolic Profile in Ex Vivo Mouse Intestinal Crypt Organoid Cultures

Real Time Analysis of Metabolic Profile in Ex Vivo Mouse Intestinal Crypt Organoid Cultures

代谢谱的实时分析<em&gt;离体</em&gt;鼠标肠隐窝类器官培养

The small intestinal mucosa exhibits a repetitive architecture organized into two fundamental structures: villi, projecting into the intestinal lumen and ...

The c-FOS Protein Immunohistological Detection: A Useful Tool As a Marker of Central Pathways Involved in Specific Physiological Responses In Vivo and Ex Vivo

The c-FOS Protein Immunohistological Detection: A Useful Tool As a Marker of Central Pathways Involved in Specific Physiological Responses In Vivo and Ex Vivo

在c-fos蛋白免疫组织学检测：一个有用​​的工具，中央通路在特定的生理反应所涉及的标记<em&gt;在体内</em&gt;和<em&gt;离体</em

Many studies seek to identify and map the brain regions involved in specific physiological regulations. The proto-oncogene c-fos, an immediate early gene, ...

In Vitro and In Vivo Approaches to Determine Intestinal Epithelial Cell Permeability

In Vitro and In Vivo Approaches to Determine Intestinal Epithelial Cell Permeability

肠上皮细胞通透性的体外及体内测定方法

The intestinal barrier defends against pathogenic microorganism and microbial toxin. Its function is regulated by tight junction permeability and ...

Forced Salivation As a Method to Analyze Vector Competence of Mosquitoes

强迫流涎法分析蚊虫的载体能力

Vector competence is defined as the potential of a mosquito species to transmit a mosquito-borne virus (mobovirus) to a vertebrate host. Viable virus ...