Name: an ectopic chemokine expression model for testing macrophage recruitment in vivo
Uploaded: 2019-09-25T14:30:00
Description: Watch this Scientific Journal Video about An Ectopic Chemokine Expression Model for Testing Macrophage Recruitment In Vivo at JoVE.com

我们感谢彭景荣博士分享Tg（fabp10a：DsRed）转基因线;温子龙博士分享Tg（mpeg1：GFP）转基因线;川口孝一博士提供pTol2向量。这项工作得到了国家自然科学基金（31771594）、广东省科技计划项目（2019A030317001）和中央高校基础研究基金（D2191450）的支持。

注：所有样品均通过苯基尿酸（PTU）蛋水处理，以抑制色素。
1. Tg的生成 （fabp10a： il34） 转基因构造和鱼类注射
<ol><li>将斑马鱼的 2.8 kb fabp10a启动子8和 IL-34 编码区域 （ENSDART00000126460.3） 克隆到 pTol2 矢量中，以生成 fabp10a-il34 构造。将结构与转座酶mRNA一起注入单细胞阶段Tg（mpeg1：GFP）和WT鱼胚胎。 将fabp10a-i...

<ol>
	<li>Howe, K., 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=The+zebrafish+reference+genome+sequence+and+its+relationship+to+the+human+genome.">The zebrafish reference genome sequence and its relationship to the human genome.</a> Nature. 496 (7446), 498-503 (2013).</li><li>Wang, Y., 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=IL-34+is+a+tissue-restricted+ligand+of+CSF1R+required+for+the+development+of+Langerhans+cells+and+microglia.">IL-34 is a tissue-restricted ligand of CSF1R required for the development of Langerhans cells and microglia.</a> Nature Immunology. 13 (8), 753-760 (2012).</li><li>Lin, 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=Discovery+of+a+cytokine+and+its+receptor+by+functional+screening+of+the+extracellular+proteome.">Discovery of a cytokine and its receptor by functional screening of the extracellular proteome.</a> Science. 320 (5877), 807-811 (2008).</li><li>Wei, 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=Functional+overlap+but+differential+expression+of+CSF-1+and+IL-34+in+their+CSF-1+receptor-mediated+regulation+of+myeloid+cells.">Functional overlap but differential expression of CSF-1 and IL-34 in their CSF-1 receptor-mediated regulation of myeloid cells.</a> Journal of leukocyte biology. 88 (3), 495-505 (2010).</li><li>Etienne, D., Foucher, S. B. L. P., Norbert Ifrah, P. G. Y. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-34+Induces+the+Differentiation+of+Human+Monocytes+into+Immunosuppressive+Macrophages.+Antagonistic+Effects+of+GM-CSF+and+IFNc.">IL-34 Induces the Differentiation of Human Monocytes into Immunosuppressive Macrophages. Antagonistic Effects of GM-CSF and IFNc.</a> PLoS One. 8 (2), e56045 (2013).</li><li>Segaliny, A. I., 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=Syndecan-1+regulates+the+biological+activities+of+interleukin-34.">Syndecan-1 regulates the biological activities of interleukin-34.</a> Biochimica et Biophysica Acta. 1853 (5), 1010-1021 (2015).</li><li>Zhou, S. L., 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=miR-28-5p-IL-34-macrophage+feedback+loop+modulates+hepatocellular+carcinoma+metastasis.">miR-28-5p-IL-34-macrophage feedback loop modulates hepatocellular carcinoma metastasis.</a> Hepatology. 63 (5), 1560-1575 (2016).</li><li>Gordon, J. I., 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=Tissue+specific+expression+and+developmental+regulation+of+two+genes+coding+for+rat+fatty+acid+binding+proteins.">Tissue specific expression and developmental regulation of two genes coding for rat fatty acid binding proteins.</a> Journal of Biological Chemistry. 260 (4), 1995-1998 (1985).</li><li>Westerfield, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish (Danio rerio). , (2000).</li><li>N&#252;sslein-Volhard, C., Dahm, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Zebrafish: a practical approach. , (2002).</li><li>Jiang, Y., Chen, J., Yen, K., Xu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ectopically+Expressed+IL-34+Can+Efficiently+Induce+Macrophage+Migration+to+the+Liver+in+Zebrafish.">Ectopically Expressed IL-34 Can Efficiently Induce Macrophage Migration to the Liver in Zebrafish.</a> Zebrafish. 16 (2), 165-170 (2019).</li></ol>

这里描述的协议允许我们研究化学对宏噬菌体体内行为的功能，并且该过程需要一些技术专长。总之，在协议中有几个关键步骤可以避免并发症：1） 选择一条合适的转基因线，显示特定且强的转基因信号，为感兴趣的细胞贴上标签;2） 选择适合成像和转基因基因过度表达的组织;3） 进行敏感和特异性的RNA探针;4） 选择适当的观察时间窗口，以准确捕获细胞行为。
在全坐式荧...

斑马鱼是一种原产于印度的热带硬骨淡水鱼。在基因保护方面，斑马鱼与人类1的相似度为87%。通过研究斑马鱼的基因调节、蛋白质功能和细胞行为，如迁徙、增殖et.al等，我们可以对人类的相关课题进行见解。斑马鱼胚胎可用于观察抑制色素后不同阶段早期胚胎的发育。同时，斑马鱼只需三个月就能发育成性成熟，斑马鱼每4天就能产下数百个卵。体积小，繁殖简单，繁殖能力强，这些优势使斑马鱼养殖非常节省空间，有利于大规模养殖。传统的哺乳动物模型小鼠的维护成本比斑马鱼高，因此限制了小鼠饲养的规模。在早期胚胎发育方面，由于母子宫小鼠胚胎发育的特点，小鼠胚胎在活体条件下难以观察。相反，斑马鱼胚胎在外部发育和透明，因此在显微镜下很容易观察。此外，斑马鱼很容易为相关基因功能研究构建各种转基因线。目前，各种斑马鱼转基因线可用于标记不同类型的细胞。现在，在特定位置构造转基因线来过度表达化学素，并研究化学素对斑马鱼细胞行为的作用是非常方便的。
在这里，我们提供了一个工作流程，使用斑马鱼转基因线来研究IL-34对体内巨噬细胞行为2，3，4，5，6，7的功能。首先，我们构建了基因il34的肝脏特异性过度表达质粒，并将质粒注入单细胞阶段Tg（mpeg1：GFP）鱼胚胎中，该胚胎通过荧光蛋白GFP专门标记巨噬细胞。然后，我们使用全座荧光原位杂交和免疫染色来检测il34表达的图案以及巨噬细胞的数量或位置。注射的WT胚胎被培养成稳定的转基因线。在这些步骤中，我们建立并验证了细胞因子生成线，并直观地评估了对巨噬细胞分布的影响。最后，为了研究巨噬细胞行为对细胞因子的反应，我们使用共聚焦活成像直接观察巨噬细胞迁移，以确认il34对体内巨噬细胞迁移的功能。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Antibody</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Alexa 488-Anti-Goat antibody</td>
			<td>Invitrogen</td>
			<td>A11055</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Digoxigenin-HRP&#160;</td>
			<td>perkinelmer</td>
			<td>NEF832001EA</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat-Anti-GFP antibody</td>
			<td>Abcam</td>
			<td>ab6658</td>
			<td></td>
		</tr>
		<tr>
			<td>Reagent</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CaCl2&#183; 2H2O</td>
			<td>Sigma</td>
			<td>21097</td>
			<td></td>
		</tr>
		<tr>
			<td>Cyanine 3 Plus Amplification Reagent</td>
			<td>perkinelmer</td>
			<td>NEL745001KT</td>
			<td></td>
		</tr>
		<tr>
			<td>E2 solution</td>
			<td></td>
			<td></td>
			<td>15 mM&#160; NaCl +0.5 mM KCl +1.0 mM MgSO4+150 &#181;M&#160; KH2PO4 + 50 &#181;M&#160; Na2HPO4 +1.0 mM CaCl2 + 0.7 mM NaHCO3&#160;</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum(FBS)</td>
			<td>Life</td>
			<td>10099-133</td>
			<td></td>
		</tr>
		<tr>
			<td>formamide</td>
			<td>Diamond</td>
			<td>A100314</td>
			<td></td>
		</tr>
		<tr>
			<td>glycerol&#160;</td>
			<td>Sigma</td>
			<td>V900860</td>
			<td></td>
		</tr>
		<tr>
			<td>heparin sodium</td>
			<td>Sigma</td>
			<td>H3149</td>
			<td></td>
		</tr>
		<tr>
			<td>hybridization buffer(HB)</td>
			<td></td>
			<td></td>
			<td>50% formamide+ 5&#215;SSC+9 mM sodium citrate+50 &#956;g/ml heparin sodium+ 500 &#956;g/ml tRNA+ 0.1% Tween20</td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma</td>
			<td>P5405</td>
			<td></td>
		</tr>
		<tr>
			<td>KH2PO4</td>
			<td>Sigma</td>
			<td>P5655</td>
			<td></td>
		</tr>
		<tr>
			<td>low melting agarose</td>
			<td>Sigma</td>
			<td>A9414</td>
			<td></td>
		</tr>
		<tr>
			<td>methanol</td>
			<td>GHTECH</td>
			<td>1.17112.023</td>
			<td></td>
		</tr>
		<tr>
			<td>methylene blue&#160;</td>
			<td>Sigma</td>
			<td>M9140</td>
			<td></td>
		</tr>
		<tr>
			<td>MgSO4</td>
			<td>Sigma</td>
			<td>M2643</td>
			<td></td>
		</tr>
		<tr>
			<td>Na2HPO4</td>
			<td>Sigma</td>
			<td>S5136</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma</td>
			<td>S5886</td>
			<td></td>
		</tr>
		<tr>
			<td>NaHCO3&#160;</td>
			<td>Sigma</td>
			<td>S5761</td>
			<td></td>
		</tr>
		<tr>
			<td>paraformaldehyde(PFA)</td>
			<td>Sigma</td>
			<td>158127</td>
			<td>Suspend 16 g of PFA in 400 ml of 1x PBS, heat at 60 &#176;C&#160; to dissolve about 30 min. This solution can be prepared in advance and stored at -4 &#176;C. Caution. Manipulate with mask.</td>
		</tr>
		<tr>
			<td>10&#215;PBS</td>
			<td></td>
			<td></td>
			<td>14.2 g Na2HPO4+80 g NaCl+2 g KCl+ 2.4 g KH2PO4 in 1L ddH2O</td>
		</tr>
		<tr>
			<td>phenylthiourea(PTU)</td>
			<td>Sigma</td>
			<td>P7629</td>
			<td></td>
		</tr>
		<tr>
			<td>1&#215;Plus Amplification Diluent</td>
			<td>perkinelmer</td>
			<td>NEL745001KT</td>
			<td></td>
		</tr>
		<tr>
			<td>Proteinase K&#160;</td>
			<td>Fermentas</td>
			<td>E00492</td>
			<td></td>
		</tr>
		<tr>
			<td>20&#215;Saline sodium citrate(SSC)</td>
			<td></td>
			<td></td>
			<td>175.3 g NaCl+ 88.2 g sodium citrate in 1 L ddH2O, PH 7.0</td>
		</tr>
		<tr>
			<td>sodium citrate</td>
			<td>Sigma</td>
			<td>A5040</td>
			<td></td>
		</tr>
		<tr>
			<td>tricaine</td>
			<td>Sigma</td>
			<td>E10521</td>
			<td></td>
		</tr>
		<tr>
			<td>tRNA&#160;</td>
			<td>Sigma</td>
			<td>R6625</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween20</td>
			<td>Sigma</td>
			<td>P2287</td>
			<td></td>
		</tr>
		<tr>
			<td>Plasmid</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>pBLK-fabp10a-il34-sv40</td>
			<td></td>
			<td></td>
			<td>For Tg (fab10a:il34) transgenic line generation</td>
		</tr>
		<tr>
			<td>pBSK-il34</td>
			<td></td>
			<td></td>
			<td>For il34 probe preparation</td>
		</tr>
		<tr>
			<td>Fish</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tg (mpeg1: GFP)</td>
			<td></td>
			<td></td>
			<td>Label macrophages with GFP</td>
		</tr>
		<tr>
			<td>Tg (fabp10a: DsRed)</td>
			<td></td>
			<td></td>
			<td>Label liver cells with DsRed</td>
		</tr>
		<tr>
			<td>Tg (fab10a:il34)</td>
			<td></td>
			<td></td>
			<td>Over-expression IL-34 in liver cells</td>
		</tr>
	</tbody>
</table>

an ectopic chemokine expression model for testing macrophage recruitment in vivo

斑马鱼在基础和生物医学研究中被广泛使用。许多斑马鱼转基因线目前可用于标记各种类型的细胞。由于斑马鱼的胚胎体透明，研究一种化学素对体内某类细胞行为的影响是很方便的。在这里，我们提供了一个工作流程，以调查化学体内巨噬菌体迁移的功能。我们构建了一个组织特异性过度表达质粒，以过度表达IL-34，并将质粒注入单细胞级转基因鱼胚胎中，其巨噬细胞被荧光蛋白专门标记。然后，我们使用全安装荧光原位杂交和免疫染色来检测化学体表达的模式以及巨噬细胞的数量或位置。注射的WT胚胎被培养成稳定的转基因线。最后，利用共聚焦活成像直接观察稳定转基因鱼的巨噬行为，研究IL-34对体内巨噬细胞的功能。

斑马鱼协议中涉及的步骤如图2所示。首先，我们生成了 pBLK-fabp10a-il34-sv40 构造，其中il34由fabp10a启动子驱动（图 2）。该结构被微注入单细胞阶段Tg （mpeg1：GFP）斑马鱼胚胎，可以标记巨噬细胞与GFP和WT胚胎，这些胚胎被培养成人，以产生转基因稳定线（图2）。il34的表达?...

Watch this Scientific Journal Video about An Ectopic Chemokine Expression Model for Testing Macrophage Recruitment In Vivo at JoVE.com

An Ectopic Chemokine Expression Model for Testing Macrophage Recruitment In Vivo

为了测试化学金对体内巨噬细胞招募的影响，使用整个原位杂交检测化学体异位表达，免疫染色用于标记巨噬细胞。实时成像用于实时观察巨噬菌体迁移。

用于测试 Vivo 中巨噬菌体招募的异位化学表达模型

Zebrafish are widely used in basic and biomedical research. Many zebrafish transgenic lines are currently available to label various types of cells. Owing ...

此处描述的协议允许我们研究化疗的功能和巨噬细胞在体内的行为。现有的大多数细胞化疗实验模型都是基于体外细胞实验。但是体外实验有时太简单，不能对体内的复杂环境进行建模。此外，它们可能并不代表体内的化疗能力。这些方法利用斑马鱼的具体优势进行直接细胞行为观察，这对小鼠来说是困难的。首先生成 TGFABP-10A 白细胞间-34 转基因结构。将FABP-10A白细胞素-34构造注入一个细胞阶段Tg（mpeg1：GFP）转基因和野生型鱼胚胎以及转相 MRNA。根据手稿说明饲养和收集胚胎。然后，在4摄氏度下过夜或室温下用4%甲醛固定两小时。固定后，用PBST清洗胚胎三次，每次洗涤五分钟。在PBST中用50%甲醇单独脱水胚胎，然后用100%甲醇，然后换至新鲜的100%甲醇，并在负20摄氏度下储存至少2小时。当准备好探针杂交时，在PBST中50%甲醇中单独补充胚胎的水分。然后，用 PBST 清洗三次，每次洗涤五分钟。在室温下用PBST中的蛋白酶K消化胚胎。然后，丢弃消化液，在室温下用4%甲醛重复固定20分钟。固定后，用PBST清洗胚胎两次，每次洗涤10分钟。在 65 摄氏度下使用加热杂交缓冲液进行预润滑，至少一小时。然后，将白细胞间-34探针预热至65摄氏度，10分钟。将杂交缓冲液回收到原始管中，并在 65 摄氏度下使用预热探头进行杂交。第二天，用50%的甲酰胺和2X SSCT清洗胚胎，然后用2XSSCT清洗，然后在65摄氏度下清洗0.2XSSCT。每次洗涤重复三次，每次洗涤20分钟。然后，用PBST清洗胚胎三次，每次洗涤五分钟。在室温下用600毫升的阻滞缓冲液阻挡样品一小时。然后，加入400微升的抗迪戈西格宁HRP抗体溶液，并在4摄氏度下孵育胚胎过夜。第二天，取出抗体，用PBST洗六次胚胎，每次洗涤20分钟。最后一次洗涤后，用30微升1X加扩增稀释剂冲洗每个样品5分钟。在黑暗中孵育氟磷泰胺工作溶液中的样品5至15分钟。如果信号较弱，将潜伏时间延长到30分钟。然后，用PBST清洗胚胎三次，每次洗涤10分钟。并在一夜之间用四摄氏度的原抗体孵育它们。第二天，用PBST洗胚5次，每次洗涤30分钟，并在4摄氏度下用二级抗体孵育。第二天，在PBST中清洗胚胎三次，每次洗涤10分钟，并在4摄氏度或负20摄氏度的黑暗中储存在70%甘油中，时间更长。在成像之前，使用荧光显微镜选择 DS 红色和 GFP 双阳性胚胎。要安装鱼，使用金属浴将1%低熔化的气糖加热至90摄氏度以上。然后，冷却到体温，混合在50微升0.2%三丁。将麻醉胚胎转移到底部盖滑梯上的小盘子。去除周围的水，慢慢将低熔化的气糖滴在胚胎上。在加糖凝固之前，请仔细设置鱼的位置，使肝脏区域靠近盖滑梯。一旦阿加罗斯凝固，小心地用另一层加糖盖住它，以加固它。将盘子放在共和显微镜载体桌上，用三辛的 E2 溶液盖住鱼，然后开始成像。要操作共体显微镜，请打开 Zen Black 2.3 软件并单击定位，然后孵育，然后将温度设置为 29 摄氏度。单击采集菜单，在智能设置菜单中选择所需的扫描模式和激光。然后选择 Z 堆栈和位置。单击实验设计器菜单并选择在第一个块中启用多块实验，以在低放大率下查找样本。然后，切换到高放大率，让观测区域位于视场的中心。设置位置和 Z 堆栈信息。然后，选择适当的激光强度、扫描层和成像速度。设置所有块后，设置适当的循环数并开始录制。该协议用于将肝脏特异性白细胞-34通过表达质粒注射到转基因鱼胚胎中，转基因鱼的巨噬细胞被贴上了GP的标签。利用全安装荧光在原位杂交和免疫污染分析白细胞间-34和GGFP标记巨噬细胞的表达。在未注射的细胞数量上对巨噬细胞数进行了定量分析，并构建了注射胚胎的肝和尾区。活成像用于直接观察巨噬细胞，标有绿色，通过控制鱼的肝脏，并迁移到白细胞间-34的肝脏，过度表达鱼。该协议的重要步骤包括选择合适的转基因生产线来标记感兴趣的细胞。并适当的组织成像您的转基因基因表达以及适当的观察时间窗口。该技术可用于研究其他化疗素对体内其他细胞（如T细胞和嗜中性粒细胞）行为的功能。

Research

JoVE Journal

Developmental Biology

Live Imaging of Innate Immune and Preneoplastic Cell Interactions Using an Inducible Gal4/UAS Expression System in Larval Zebrafish Skin

先天免疫和癌前细胞间的相互作用使用可诱导的Gal4 / UAS表达系统在斑马鱼幼体皮肤实时成像

Here we describe a method to conditionally induce epithelial cell transformation by the use of the 4-Hydroxytamoxifen (4-OHT) inducible KalTA4-ERT2/UAS ...

科研

发育生物学

An Optogenetic Approach for Assessing Formation of Neuronal Connections in a Co-culture System

评定神经连接的形成在共培养体系的光遗传学方法

Here we describe a protocol to generate a co-culture consisting of 2 different neuronal populations. Induced pluripotent stem cells (iPSCs) are ...

Development of an Ethanol-induced Fibrotic Liver Model in Zebrafish to Study Progenitor Cell-mediated Hepatocyte Regeneration

乙醇引起的肝纤维化模型的斑马鱼发展研究祖细胞介导的​​肝细胞的再生

Sustained liver fibrosis with continuation of extracellular matrix (ECM) protein build-up results in the loss of cellular competency of the liver, leading ...

Drug Treatment and In Vivo Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

Drug Treatment and In Vivo Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

戒毒治疗和<em&gt;在体内</em&gt;在青鳉鱼骨质疏松模型成骨破骨细胞相互作用的成像

Bone-forming osteoblasts interact with bone-resorbing osteoclasts to coordinate the turnover of bone matrix and to control skeletal homeostasis. Medaka ...

An Ex Vivo Method for Time-Lapse Imaging of Cultured Rat Mesenteric Microvascular Networks

An Ex Vivo Method for Time-Lapse Imaging of Cultured Rat Mesenteric Microvascular Networks

一个<em&gt;离体</em&gt;方法培养大鼠肠系膜微血管网络的时间推移成像

Angiogenesis, defined as the growth of new blood vessels from pre-existing vessels, involves endothelial cells, pericytes, smooth muscle cells, immune ...

In Vivo Imaging of Transgenic Gene Expression in Individual Retinal Progenitors in Chimeric Zebrafish Embryos to Study Cell Nonautonomous Influences

In Vivo Imaging of Transgenic Gene Expression in Individual Retinal Progenitors in Chimeric Zebrafish Embryos to Study Cell Nonautonomous Influences

在嵌合斑马鱼胚胎的个别视网膜祖细胞转基因表达的体内成像来研究细胞非自治的影响

The genetic and technical strengths have made the zebrafish vertebrate a key model organism in which the consequences of gene manipulations can be traced ...

Establishment of Larval Zebrafish as an Animal Model to Investigate Trypanosoma cruzi Motility In Vivo

Establishment of Larval Zebrafish as an Animal Model to Investigate Trypanosoma cruzi Motility In Vivo

斑马鱼幼虫动物模型的建立研究锥虫运动性体内

Chagas disease is a parasitic infection caused by Trypanosoma cruzi, whose motility is not only important for localization, but also for cellular binding ...

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans

rna 干扰和热休克诱导转录因子表达在重编程生殖细胞到神经元中的应用C. 线虫

Studying the cell biological processes during converting the identities of specific cell types provides important insights into mechanism that maintain ...

Culturing of Retinal Pigment Epithelial Cells on an Ex Vivo Model of Aged Human Bruch's Membrane

Culturing of Retinal Pigment Epithelial Cells on an Ex Vivo Model of Aged Human Bruch's Membrane

老年人刷膜的体外模型对视网膜色素上皮细胞的培养

Aside from vitamins and antioxidants recommended by the Age-Related Eye Disease Study, there is no effective therapy for &#34;dry,&#34;&#160;or atrophic ...

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues

平行板灌注系统的体外模型研究了细菌对移植组织的粘附性

Various valved conduits and stent-mounted valves are used for right ventricular outflow tract (RVOT) valve replacement in patients with congenital heart ...