Name: electroporation method for in vivo delivery of plasmid dna in the adult zebrafish telencephalon
Uploaded: 2019-09-13T15:00:00
Description: Watch this Scientific Journal Video about Electroporation Method for In Vivo Delivery of Plasmid DNA in the Adult Zebrafish Telencephalon at JoVE.com

特别感谢詹姆斯·科普蒂编辑手稿。我们还感谢德国研究基金会 （DFG） 向 JN 提供的资金，由 SFB 870 和 SPP"嗅觉综合分析"和 SPP 1738"非编码 RNA 在神经系统开发、可塑性和疾病中的新作用"，SPP1757在慕尼黑系统神经学集群（EXC 2145 SyNergy – ID 390857198）框架内的胶质异质性"和卓越战略。

本议定书中使用的所有动物均按标准饲养条件饲养，实验均按照欧盟和上巴伐利亚州政府的处理准则和条例进行（AZ 55.2-1-54-2532-0916）。
1. 用于电穿孔的质粒混合物的制备
<ol><li>稀释无菌水中感兴趣的质粒，并加入快速的绿色染色剂溶液[1毫克/mL]。确保质粒的最终浓度为±1 μg/μL。以不超过3%的浓度添加污渍，因为它的唯一目的是给溶液着色和可视化心室注射。</li>
	<li>制备?...

<ol>
	<li>Kaslin, J., Ganz, J., Brand, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proliferation,+neurogenesis+and+regeneration+in+the+non-mammalian+vertebrate+brain.">Proliferation, neurogenesis and regeneration in the non-mammalian vertebrate brain.</a> Philosophical Transactions of the Royal Society of London Series B Biological Sciences. 363 (1489), 101-122 (2008).</li><li>Baumgart, E. V., Barbosa, J. S., Bally-Cuif, L., Gotz, M., Ninkovic, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stab+wound+injury+of+the+zebrafish+telencephalon:+a+model+for+comparative+analysis+of+reactive+gliosis.">Stab wound injury of the zebrafish telencephalon: a model for comparative analysis of reactive gliosis.</a> Glia. 60 (3), 343-357 (2012).</li><li>Barbosa, 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=Live+imaging+of+adult+neural+stem+cell+behavior+in+the+intact+and+injured+zebrafish+brain.">Live imaging of adult neural stem cell behavior in the intact and injured zebrafish brain.</a> Science. 346 (6236), 789-793 (2015).</li><li>Kishimoto, N., Shimizu, K., Sawamoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+regeneration+in+a+zebrafish+model+of+adult+brain+injury.">Neuronal regeneration in a zebrafish model of adult brain injury.</a> Disease Model and Mechanisms. 5 (2), 200-209 (2012).</li><li>Kroehne, V., Freudenreich, D., Hans, S., Kaslin, J., Brand, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+of+the+adult+zebrafish+brain+from+neurogenic+radial+glia-type+progenitors.">Regeneration of the adult zebrafish brain from neurogenic radial glia-type progenitors.</a> Development. 138 (22), 4831-4841 (2011).</li><li>Marz, M., Schmidt, R., Rastegar, S., Strahle, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regenerative+response+following+stab+injury+in+the+adult+zebrafish+telencephalon.">Regenerative response following stab injury in the adult zebrafish telencephalon.</a> Developmental Dynamics. 240 (9), 2221-2231 (2011).</li><li>Ayari, B., Elhachimi, K. H., Yanicostas, C., Landoulsi, A., Soussi-Yanicostas, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prokineticin+2+expression+is+associated+with+neural+repair+of+injured+adult+zebrafish+telencephalon.">Prokineticin 2 expression is associated with neural repair of injured adult zebrafish telencephalon.</a> Journal of Neurotrauma. , (2010).</li><li>Skaggs, K., Goldman, D., Parent, J. 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A., Talbot, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial+cell+development+and+function+in+zebrafish.">Glial cell development and function in zebrafish.</a> Cold Spring Harbor Perspectives in Biology. 7 (2), a020586 (2014).</li><li>Obermann, 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=The+Surface+Proteome+of+Adult+Neural+Stem+Cells+in+Zebrafish+Unveils+Long-Range+Cell-Cell+Connections+and+Age-Related+Changes+in+Responsiveness+to+IGF.">The Surface Proteome of Adult Neural Stem Cells in Zebrafish Unveils Long-Range Cell-Cell Connections and Age-Related Changes in Responsiveness to IGF.</a> Stem Cell Reports. 12 (2), 258-273 (2019).</li><li>Progatzky, F., Dallman, M. J., Lo Celso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+seeing+to+believing:+labelling+strategies+for+in+vivo+cell-tracking+experiments.">From seeing to believing: labelling strategies for in vivo cell-tracking experiments.</a> Interface Focus. 3 (3), 20130001 (2013).</li><li>Kusumanto, Y. 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=Improvement+of+in+vivo+transfer+of+plasmid+DNA+in+muscle:+comparison+of+electroporation+versus+ultrasound.">Improvement of in vivo transfer of plasmid DNA in muscle: comparison of electroporation versus ultrasound.</a> Drug Delivery. 14 (5), 273-277 (2007).</li><li>Zou, M., De Koninck, P., Neve, R. L., Friedrich, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+gene+transfer+into+the+adult+zebrafish+brain+by+herpes+simplex+virus+1+(HSV-1)+and+electroporation:+methods+and+optogenetic+applications.">Fast gene transfer into the adult zebrafish brain by herpes simplex virus 1 (HSV-1) and electroporation: methods and optogenetic applications.</a> Frontiers in Neural Circuits. 8 (41), (2014).</li><li>Van Tendeloo, V. F., 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=Highly+efficient+gene+delivery+by+mRNA+electroporation+in+human+hematopoietic+cells:+superiority+to+lipofection+and+passive+pulsing+of+mRNA+and+to+electroporation+of+plasmid+cDNA+for+tumor+antigen+loading+of+dendritic+cells.">Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells.</a> Blood. 98 (1), 49-56 (2001).</li><li>Mars, 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=Electrotransfection+and+lipofection+show+comparable+efficiency+for+in+vitro+gene+delivery+of+primary+human+myoblasts.">Electrotransfection and lipofection show comparable efficiency for in vitro gene delivery of primary human myoblasts.</a> The Journal of Membrane Biology. 248 (2), 273-283 (2015).</li><li>Barbosa, J. S., Di Giaimo, R., Gotz, M., Ninkovic, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-cell+in+vivo+imaging+of+adult+neural+stem+cells+in+the+zebrafish+telencephalon.">Single-cell in vivo imaging of adult neural stem cells in the zebrafish telencephalon.</a> Nature Protocols. 11 (8), 1360-1370 (2016).</li><li>Di Giaimo, R., 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+Aryl+Hydrocarbon+Receptor+Pathway+Defines+the+Time+Frame+for+Restorative+Neurogenesis.">The Aryl Hydrocarbon Receptor Pathway Defines the Time Frame for Restorative Neurogenesis.</a> Cell Reports. 25 (12), 3241-3251 (2018).</li><li>Breunig, C. 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=One+step+generation+of+customizable+gRNA+vectors+for+multiplex+CRISPR+approaches+through+string+assembly+gRNA+cloning+(STAgR).">One step generation of customizable gRNA vectors for multiplex CRISPR approaches through string assembly gRNA cloning (STAgR).</a> PLoS One. 13 (4), e0196015 (2018).</li><li>Barbosa, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> In vivo single cell analysis reveals distinct behavior of neural stem and progenitor cells in homeostasis and regeneration in the adult brain. , (2014).</li><li>Kelsh, R. N., 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=Zebrafish+pigmentation+mutations+and+the+processes+of+neural+crest+development.">Zebrafish pigmentation mutations and the processes of neural crest development.</a> Development. 123, 369-389 (1996).</li><li>Torper, O., 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=In+Vivo+Reprogramming+of+Striatal+NG2+Glia+into+Functional+Neurons+that+Integrate+into+Local+Host+Circuitry.">In Vivo Reprogramming of Striatal NG2 Glia into Functional Neurons that Integrate into Local Host Circuitry.</a> Cell Reports. 12 (3), 474-481 (2015).</li><li>Nguyen, L. T., Atobe, K., Barichello, J. M., Ishida, T., Kiwada, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complex+formation+with+plasmid+DNA+increases+the+cytotoxicity+of+cationic+liposomes.">Complex formation with plasmid DNA increases the cytotoxicity of cationic liposomes.</a> Biological and Pharmaceutical Bulletin. 30 (4), 751-757 (2007).</li><li>Chapouton, P., Godinho, L., Detrich, H. W., Westerfield, M., Zon, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Neurogenesis. 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这种电穿孔方案是一种可靠的体内方法，用于标记单个细胞。该协议可能需要进一步适应，以标记其他细胞类型，如神经元或寡核苷酸细胞。为了获得成功的标签，可以使用含有不同促进剂的质粒。鸡-β蛋白促进剂，eF1α，CMV和泛素促进剂以前曾被用来驱动不同转基因在ependyglia及其后代23的表达。然而，观察到了不同转基因表达的动力学，这应当加以考虑。例如，CMV启动子驱动?...

斑马鱼是优秀的动物模型，用于检查刺伤后的大脑再生。与哺乳动物相比，在进化阶梯上，斑马鱼等进化较少的物种通常表现出较高的形成性神经发生率和成人神经干细胞栖息的更广区域，导致在整个过程中不断产生新的神经元成人生活中的大多数大脑区域。与哺乳动物1相比，这一特征似乎与斑马鱼的再生能力显著提升有关，因为斑马鱼在所研究的大多数脑损伤模型中具有显著的潜力，能够有效地产生新的神经元。 3，4，5，6，7，8 。在这里，斑马鱼端脑研究，因为它是一个大脑区域与突出的神经发生在成年。这些成年神经发生区与成年哺乳动物脑中神经原发生区9、10、11同源。
由于存在径向胶质细胞或脑细胞等径向胶质，斑马鱼端脑素中存在丰富的神经原发生区。Ependymoglial细胞作为常驻的成人神经干细胞，负责在完整和再生的大脑3，5中产生新的神经元。系骨追踪实验表明，心室叶膜对损伤有反应，然后增殖并产生新的神经细胞，迁移到病变部位5。由于斑马鱼端脑的电化性质，腹腔细胞排列在心室表面，并建造心室壁。背叶心室壁由背端阴膜细胞层形成（图1A）。重要的是，斑马鱼叶黄体体现了哺乳动物径向胶质和阴叶细胞的特征。长径向过程是径向胶质细胞的典型特征，而细胞延伸和紧密结（及其心室位置）是阴囊细胞12、13、14的典型特征。因此，这些细胞被称为阴叶细胞。
为了在再生过程中遵循单个阴叶细胞的体内行为，需要可靠地标记它们。荧光显微镜的体内细胞标记方法已经描述过，如内源性报告剂或亲脂染料15。与电穿孔相比，这些方法可能需要较长的时间，而且通常无法提供单细胞标记或永久长期跟踪的可能性。然而，电穿孔（除了单细胞标记）提供了将新的DNA引入宿主细胞的可能性。此外，与其他DNA移植到细胞的方法相比，电穿孔已被证明是最有效的方法之一16，17，18，19。
这里介绍的是一个电穿孔协议，已细化，以标记在成年斑马鱼端脑中的单一脑细胞。该协议允许标记单个细胞，以便长期跟踪它们20或操纵特定途径在细胞自主的方式21，22。

<table>
	<tbody>
		<tr>
			<td>Reagent/Material</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fast Green</td>
			<td>Sigma-Aldrich</td>
			<td>F7258-25G</td>
			<td>For coloring plasmid solution</td>
		</tr>
		<tr>
			<td>MS222</td>
			<td>Sigma-Aldrich</td>
			<td>A5040-25G</td>
			<td>MS222 should be stored at RT (up to two weeks) and protected from light</td>
		</tr>
		<tr>
			<td>Ultrasound gel</td>
			<td>SignaGel, Parker laboratories INC.</td>
			<td>15-60</td>
			<td>Electrode Gel</td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Air pump</td>
			<td>TetraTec APS 50, 10l-60l</td>
			<td></td>
			<td>Can be bought in the pet shops</td>
		</tr>
		<tr>
			<td>BTX Tweezertrodes Electrodes</td>
			<td>Platinum Tweezertrode, BTX Harvard Apparatus</td>
			<td>45-0486</td>
			<td>1mm diameter</td>
		</tr>
		<tr>
			<td>Electroporation device</td>
			<td>BTX ECM830 Square Wave Electroporation System, BTX Harvard Apparatus</td>
			<td>45-0662</td>
			<td></td>
		</tr>
		<tr>
			<td>Injection device</td>
			<td>FemtoJet 4i, Eppendorf</td>
			<td>5252000013</td>
			<td></td>
		</tr>
		<tr>
			<td>Standard Wall Borosillicate Glass Capillary</td>
			<td>Warner Instruments</td>
			<td>64-0766</td>
			<td>Model No: G100-4</td>
		</tr>
		<tr>
			<td>Microloader tips</td>
			<td>Eppendorf</td>
			<td>5242956003</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-knife</td>
			<td>Fine Science Tools</td>
			<td>10056-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Joystick micromanipulator</td>
			<td>Narishige Japan</td>
			<td>MN - 151</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>FemtoJet 4i, Eppendorf</td>
			<td>5252000013</td>
			<td>Needle holder comes together with the injection device</td>
		</tr>
		<tr>
			<td>Needle pulling device</td>
			<td>Narishige Japan</td>
			<td>Model No: PC-10</td>
			<td>The PC-10 was discontinued by Narishige in 2017 and replaced by the PC-100</td>
		</tr>
		<tr>
			<td>Petri dishes</td>
			<td>Greiner Bio-One International</td>
			<td>633161</td>
			<td></td>
		</tr>
	</tbody>
</table>

electroporation method for in vivo delivery of plasmid dna in the adult zebrafish telencephalon

电穿孔是一种转染方法，其中电场应用于细胞，在细胞膜中产生临时孔隙，提高渗透性，从而允许将不同的分子引入细胞。本文采用电穿孔技术将质粒引入脑膜细胞，使成年斑马鱼的腹室区域划线。这些细胞的一小部分显示了干细胞的特性，并在斑马鱼大脑中产生新的神经元;因此，研究他们的行为对于确定他们在神经发生和再生中的作用至关重要。通过电穿孔引入质粒，可以长期标记和跟踪单个细胞。此外，Cre重组酶或Cas9等质粒可输送到单个细胞，从而实现基因重组或基因编辑，并为在受控、自然中评估细胞的自主基因功能提供了独特的机会环境。最后，采用这种详细的分步电穿孔方案，成功地将质粒引入大量单细胞。

所述电穿孔方法允许将质粒DNA输送到脑细胞中，这些细胞表面位于斑马鱼端脑素和背端脑细胞层（图1A）。
如果电穿孔的结果为阳性，可以在其他阴极细胞中观察到标记的单阴白细胞（图2A，B中的红细胞）（图2A，B中为白色）。根据电穿孔过程的效率，可以标记更高（<strong class=...

Watch this Scientific Journal Video about Electroporation Method for In Vivo Delivery of Plasmid DNA in the Adult Zebrafish Telencephalon at JoVE.com

Electroporation Method for In Vivo Delivery of Plasmid DNA in the Adult Zebrafish Telencephalon

这里介绍的是一种在成年斑马鱼端脑中进行质粒DNA传递和脑细胞标记的电穿孔方法。该协议是一种快速而有效的方法，用于可视化和跟踪单个细胞，并开辟了将电穿孔应用于各种基因操作的新的可能性。

成年斑马鱼端脑中疟原虫DNA在活体传递的电穿孔方法

Electroporation is a transfection method in which an electrical field is applied to cells to create temporary pores in a cell membrane and increase its ...

该协议允许在斑马鱼电头对单个环状细胞进行标记，以便进行长期监测，以及以细胞自主的方式操作特定通路。该技术的主要优点是，它允许以快速、高效的方式进行单细胞标记，以及以细胞自主方式进行基因编辑和信号通路操作。该方法允许研究基本的细胞生物学问题，例如，关于细胞分层，上皮平衡的维持，和细胞分裂对称性。首先使用拔针器具来准备玻璃毛细管。使用微加载器尖端，用十微升质粒溶液填充准备好的玻璃毛细管。然后按菜单改变注射装置上的毛细管"，让针头加载到针架上。然后将注射装置从更换毛细管切换到注射模式。然后，使用放大倍率为四倍的立体显微镜和 Finden 钳子，只切割毛细管的尖端。然后对脚踏板施加压力，确认质粒溶液很容易从针头中跑出来，不受阻碍。当针头准备好时，将斑马鱼从饲养罐转移到麻醉液容器中。等几分钟，直到刺的运动消退。将静静鱼背侧压在立体显微镜下的预湿海绵上，使用具有 40 毫米切削刃和 0.5 毫米厚度的不锈钢解剖小刀，在遥脑后侧的鱼头骨上小心地创建一个小孔，就在配镜边界旁边。必要时倾斜鱼，将玻璃毛细管的尖端以正确的角度向头骨倾斜，以方便毛细管尖端穿过孔。然后，非常小心地将毛细管的尖端插入孔中，通过背膜状细胞层，直到它到达心灵心室，并施加压力到脚踏板约10秒，以提供约1微升的质粒溶液。通过观察绿色液体在整个心室的扩散，可以确认分娩的成功。用少量的超声波凝胶覆盖鱼电脑。对于质粒注射动物的电穿孔，从注射装置上取出海绵，将电尖的内侧浸入超声波凝胶中。将鱼头放在电极之间，正极在鱼头的腹侧，负极放在后侧。然后，在海绵中仍然将鱼体固定，轻轻精确地按压电脑，然后用脚踏板管理电流，将电极固定到位，直到所有五个脉冲都完成。如果电穿孔成功，可以在非质粒标记的电镀细胞中观察到标有单质肌细胞的质粒标记。根据电穿孔过程的效率，可以标记更高或更低的倍数电镀细胞。然而，所演示的协议产生的标签单元格数量比之前发布的协议多。请注意，标记细胞的最高密度往往主要出现在两个半球的内心室侧，因为注射的质粒液体在半球之间分布的方式。在这段视频中，斑马鱼的一个半球以3D显示，其中具有无线电过程的斑马细胞在从侧面观察到时具有品红色。通过与另一个质粒（标记核）进行共电，可以观察到细胞分裂。不成功的电穿孔导致非常低的数量或没有标记的环质细胞。然而，多生细胞最有可能被标记。它们的母体在长方体中更大，并且它们不具有从 ependymoglial 细胞层的侧视图看的无线电拉长过程。tdTomato-mem标记的细胞很可能是 ependym 细胞，它们位于 ependymoglia 层之上。相比之下，在单个细胞中引入表示质粒的 tdTomato-mem，除了初始细胞标记外，还会导致 tdTomato-mem 表达。要记住的最重要的事情是只切毛细管的尖端，并在注射时穿透腹皮层，使液体在整个心室扩散。该技术通过体内成像技术可以长期监测单个细胞，从而确立其在大脑再生中的作用及其庞大的体内遗传联系。

Research

JoVE Journal

Developmental Biology

Kidney Regeneration in Adult Zebrafish by Gentamicin Induced Injury

肾再生的成年斑马鱼庆大霉素性损伤

The kidney is essential for fluid homeostasis, blood pressure regulation and filtration of waste from the body. The fundamental unit of kidney function is ...

科研

发育生物学

Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye

斑马鱼幼虫骨骼肌诚信与伊文思蓝分析

The zebrafish model is an emerging system for the study of neuromuscular disorders.  In the study of neuromuscular diseases, the integrity of the muscle ...

Gene Transfer into the Chicken Auditory Organ by In Ovo Micro-electroporation

Gene Transfer into the Chicken Auditory Organ by In Ovo Micro-electroporation

基因转移到鸡听觉器官通过<em&gt;在卵</em&gt;微电

Chicken embryos are ideal model systems for studying embryonic development as manipulations of gene function can be conducted with relative ease in ovo. ...

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 ...

In Ovo Electroporation in the Chicken Auditory Brainstem

In Ovo Electroporation in the Chicken Auditory Brainstem

在奥运电击鸡听觉脑干中

Electroporation is a method that introduces genes of interest into biologically relevant organisms like the chicken embryo. It is long established that ...

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

成人果蝇脑内蘑菇体和感光神经元的解剖和荧光染色

Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the ...

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 ...

Nanoparticle-mediated siRNA Gene-silencing in Adult Zebrafish Heart

纳米粒子介导的 siRNA 基因沉默在斑马鱼心脏中的表达

Mammals have a very limited capacity to regenerate the heart after myocardial infarction. On the other hand, the adult zebrafish regenerates its heart ...

Visualizing the Node and Notochordal Plate In Gastrulating Mouse Embryos Using Scanning Electron Microscopy and Whole Mount Immunofluorescence

用扫描电镜和全贴免疫荧光技术可视化小鼠胚胎中的节点和外型板

The post-implantation mouse embryo undergoes major shape changes after the initiation of gastrulation and morphogenesis. A hallmark of morphogenesis is ...

Adult Zebrafish Injury Models to Study the Effects of Prednisolone in Regenerating Bone Tissue

成人斑马鱼损伤模型研究泼尼松对再生骨组织的影响

Zebrafish are able to regenerate various organs, including appendages (fins) after amputation. This involves the regeneration of bone, which regrows ...