Name: double in utero electroporation to target temporally and spatially separated cell populations
Uploaded: 2020-06-14T13:00:00
Description: Watch this Scientific Journal Video about Double In Utero Electroporation to Target Temporally and Spatially Separated Cell Populations at JoVE.com

作者感谢克里斯蒂娜·安德烈斯·卡内尔和瓦伦西亚大学动物护理设施的成员提供技术援助。我们还要感谢伊莎贝尔·法里亚斯和萨克拉门托·费伦的试剂，并与我们分享他们的设备。I.M.W由瓦伦西亚大学（GJIDI/2018/A/221）的加兰迪亚·尤韦尼尔合同供资，D.dA.D由伊斯兰大学（国际大学）部长资助。"FPI-PRE2018-086150"。C.Gil-Sanz 持有西班牙法语省法语省（MICINN） 的拉蒙和卡哈尔赠款（RYC-2015-19058）。这项工作资助了RYC-2015-19058和SAF2017-82880-R（MICINN）。

此处所述的程序已获得负责实验、瓦伦西亚大学动物福利和农业学院道德委员会的批准， 巴伦西亚纳大学（巴伦西亚纳大学）的德萨罗罗罗农村，《特兰西纳大学》和《欧洲实验室动物科学委员会》的准则，遵守西班牙立法第53/23/2013号《真实声明》以及欧洲议会和理事会第2010/63/欧盟指令中审查的国际实验室动物科学理事会（ICLAS）的指导方针。
注：该协议涉及两个不同的?...

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研究大脑皮层等细胞密度高的区域体内细胞相互作用是一项复杂的任务。传统方法，包括使用抗体来标记神经衰弱是不适合的，因为缺乏针对不同细胞群的特定标记。使用转基因鼠模型（特定细胞类型表示荧光蛋白）有助于可视化神经元过程，但这取决于此类模型的可用性。当试图想象两种特定细胞类型之间在感兴趣的基因扰动时相互作用的可能差异时，这项任务就更加复杂了，因为它涉及使用?...

大脑皮层是一个非常复杂和复杂的组织结构。为了实现这种程度的组织，皮质投影神经元经历复杂的发育过程，需要其时间生成，迁移到皮质板的最终目的地，以及建立短和远程连接11，2。2长期以来，研究皮质成因的经典方法基于对感兴趣的基因的敲除或敲模模型的使用。然而，这种策略，特别是使用有条件的挖空小鼠，既费时又昂贵，有时在存在遗传冗余或缺乏具体的CRE驱动因素等方面提出了其他问题。试图解决这些问题的方法之一，现在被广泛用于研究皮质发育的方法之一是在子宫电穿孔33，4。4在子宫电穿孔是一种用于体细胞转变的技术，允许在体内瞄准神经干细胞及其后代。该方法可用于通过表达荧光蛋白55、66来标记细胞，用于体内基因操作（即功能测定的增益或丧失）7、8、9，8,用于在体外分离7电质皮质和培养细胞98,8、10。此外，在电体电穿孔中，允许对目标区域进行时间和区域控制。该技术具有多种应用，已广泛应用于研究神经元迁移、干细胞分裂、神经元连通性等学科88、9、11、12。9,11,12
目前的手稿描述了在子宫电穿孔中使用一种称为双质电穿孔的电穿孔变体，用于分析不同时间和空间起源的大脑皮层细胞的相互作用。这些研究在使用鼠型时完成极其复杂，因为它们需要组合使用几种转基因系。本文中描述的协议的一些应用包括研究相邻细胞之间的紧密相互作用，以及通过远程投影研究远程细胞之间的相互作用。该方法要求在子宫电穿孔手术中进行两次独立的手术，在时间和空间上分离，以针对感兴趣的不同细胞群。这种方法的优点是有可能利用野生动物操纵一种或两种神经元的基因功能。此外，这些功能实验可以结合细胞质或膜标记荧光蛋白的表达相结合，以可视化靶细胞（包括树突和斧头）的精细形态，并分析与对照组相比细胞相互作用的可能差异（即仅标记荧光蛋白的细胞）。
此处划定的协议侧重于研究新皮质内的细胞相互作用，但这一策略也可用于研究与可用于子宫电穿孔的外皮质区域的相互作用，如亚皮质或丘脑13、14,14或其他结构中的细胞-细胞相互作用，如小脑15。不同区域的瞄准基于电极的方向和注入DNA的心室（横向、第三或第四）。通过此处描述的策略，我们可以标记大量细胞，这对于评估功能实验中连接/内伸的一般变化非常有用。然而，为了研究连接性的变化，人们可以使用子宫电穿孔的修改版本来获得更稀疏的标签和识别单细胞16。总之，双电镀是一种通用方法，它允许瞄准时空分离的细胞群，并在控制条件下或与功能实验相结合，详细研究它们的相互作用，大大降低了时间和经济成本。

<table>
	<tbody>
		<tr>
			<td>Ampicillin sodium salt</td>
			<td>Sigma-Aldrich</td>
			<td>A9518-25G</td>
			<td></td>
		</tr>
		<tr>
			<td>Aspirator tube</td>
			<td>Sigma-Aldrich</td>
			<td>A5177-5EA</td>
			<td></td>
		</tr>
		<tr>
			<td>Baby-mixter hemostat (perfusion)</td>
			<td>Fine Science Tools (FST)</td>
			<td>13013-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Borosilicate glass capillary</td>
			<td>WPI</td>
			<td>1B100-6</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine (BUPREX 0,3 mg/ml)</td>
			<td>Rb Pharmaceuticals Limited</td>
			<td>921425</td>
			<td></td>
		</tr>
		<tr>
			<td>CAG-BFP plasmid</td>
			<td>Kindly provided by U.M&#252;ller Lab</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CAG-EGFP plasmid</td>
			<td>Kindly provided by U.M&#252;ller Lab</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CAG-mCherry plasmid</td>
			<td>Kindly provided by U.M&#252;ller Lab</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CAG-mtdTomato-2A-nGFP plasmid</td>
			<td>Kindly provided by U.M&#252;ller Lab</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Confocal microscope</td>
			<td>Olympus</td>
			<td>FV10i</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Swabs</td>
			<td>BFHCVDF</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cyanoacrylate glue</td>
			<td>B. Braun Surgical</td>
			<td>1050044</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting scope</td>
			<td>Zeiss</td>
			<td>stemi 305</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont Forceps #5 Fine Forceps</td>
			<td>Fine Science Tools (FST)</td>
			<td>11254-20</td>
			<td></td>
		</tr>
		<tr>
			<td>ECM830 Square Wave Electroporator</td>
			<td>BTX</td>
			<td>45-0052</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric Razor</td>
			<td>Oster</td>
			<td>76998</td>
			<td></td>
		</tr>
		<tr>
			<td>Endotoxin-free TE buffer</td>
			<td>QIAGEN</td>
			<td>1018499</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol wipes</td>
			<td>BFHCVDF</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Extra Fine Graefe Forceps</td>
			<td>Fine Science Tools (FST)</td>
			<td>11150-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Eye ointment</td>
			<td>Alcon</td>
			<td>682542.6</td>
			<td></td>
		</tr>
		<tr>
			<td>Fast Green dye</td>
			<td>Sigma-Aldrich</td>
			<td>F7252-5G</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Scissors</td>
			<td>Fine Science Tools (FST)</td>
			<td>14069-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence LEDs</td>
			<td>CoolLED</td>
			<td>pE-300-W</td>
			<td></td>
		</tr>
		<tr>
			<td>Genopure Plasmid Maxi Kit</td>
			<td>Roche</td>
			<td>3143422001</td>
			<td></td>
		</tr>
		<tr>
			<td>Halsted-Mosquito Hemostats (suture)</td>
			<td>Fine Science Tools (FST)</td>
			<td>91308-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating Pad</td>
			<td>UFESA</td>
			<td>AL5514</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted epifluorescence microscope</td>
			<td>Nikon</td>
			<td>Eclipse TE2000-S</td>
			<td></td>
		</tr>
		<tr>
			<td>Iodine wipes</td>
			<td>Lorsoul</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isofluorane vaporizer</td>
			<td>Flow-Meter</td>
			<td>A15B5001</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Karizoo</td>
			<td>586259</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine (Anastemine)</td>
			<td>Fatro Ib&#233;rica SL</td>
			<td>583889-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Kimtech precision wipes</td>
			<td>Kimberly-Clark</td>
			<td>7252</td>
			<td></td>
		</tr>
		<tr>
			<td>LB (Lennox) Agar GEN</td>
			<td>Labkem</td>
			<td>AGLB-00P-500</td>
			<td></td>
		</tr>
		<tr>
			<td>LB (Lennox) broth GEN</td>
			<td>Labkem</td>
			<td>LBBR-00P-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Low-melting point agarose</td>
			<td>Fisher Scientific</td>
			<td>BP165-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Medetomidine (Sedator)</td>
			<td>Dechra</td>
			<td>573749.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope coverslips</td>
			<td>Menel-Gl&#228;ser</td>
			<td>15747592</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope Slides</td>
			<td>Labbox</td>
			<td>SLIB-F10-050</td>
			<td></td>
		</tr>
		<tr>
			<td>Mounting medium</td>
			<td>Electron Microscopy Sciences</td>
			<td>17984-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Mutiwell plates (24)</td>
			<td>SPL Life Sciences</td>
			<td>32024</td>
			<td></td>
		</tr>
		<tr>
			<td>Mutiwell plates (48)</td>
			<td>SPL Life Sciences</td>
			<td>32048</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl (for saline solution)</td>
			<td>Fisher Scientific</td>
			<td>10112640</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle 25 G (BD Microlance 3)</td>
			<td>Becton, Dickinson and Company</td>
			<td>300600</td>
			<td></td>
		</tr>
		<tr>
			<td>Orbital incubator S150</td>
			<td>Stuart Scientific</td>
			<td>5133</td>
			<td></td>
		</tr>
		<tr>
			<td>P Selecta Incubator</td>
			<td>J. P. Selecta, s.a.</td>
			<td>0485472</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>PanReac AppliedChem</td>
			<td>A3813</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin</td>
			<td>Sigma -Aldrich</td>
			<td>P4333</td>
			<td></td>
		</tr>
		<tr>
			<td>Peristaltic perfusion pump</td>
			<td>Cole-Parmer</td>
			<td>EW-07522-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Platinum Tweezertrode, 5 mm Diameter</td>
			<td>Btx</td>
			<td>45-0489</td>
			<td></td>
		</tr>
		<tr>
			<td>Reflex Skin Closure System - 7mm Clips, box of 100</td>
			<td>AgnThos</td>
			<td>203-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Reflex Skin Closure System - Clip Applyer, 7mm</td>
			<td>AgnThos</td>
			<td>204-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring Forceps</td>
			<td>Fine Science Tools (FST)</td>
			<td>11103-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium azide</td>
			<td>PanReac AppliedChem</td>
			<td>122712-1608</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical absorbent pad (steryle)</td>
			<td>HK Surgical</td>
			<td>PD-M</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture (Surgicryl PGA 6-0)</td>
			<td>SMI Suture Materials</td>
			<td>BYD11071512</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe 1ml (BD plastipak)</td>
			<td>Becton, Dickinson and Company</td>
			<td>303172</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue Culture Dish 100 x 20 mm</td>
			<td>Falcon</td>
			<td>353003</td>
			<td></td>
		</tr>
		<tr>
			<td>Vertical Micropipette Puller</td>
			<td>Sutter Instrument Co</td>
			<td>P-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Vertical microscope</td>
			<td>Nikon</td>
			<td>Eclipse Ni</td>
			<td></td>
		</tr>
		<tr>
			<td>Vibratome</td>
			<td>Leica</td>
			<td>VT1200S</td>
			<td></td>
		</tr>
	</tbody>
</table>

double in utero electroporation to target temporally and spatially separated cell populations

在子宫电穿孔是一种体内DNA转移技术，广泛用于研究哺乳动物皮质成基的分子和细胞机制。此过程利用大脑心室，允许引入感兴趣的DNA，并使用一对电极引导遗传物质进入心室内衬的细胞，神经干细胞。这种方法允许研究人员标记所需的细胞和/或操作这些细胞中感兴趣的基因的表达。它有多种应用，包括针对神经元迁移、系系跟踪和斧头路径查找的测定。这种方法的一个重要特点是它的时间和区域控制，允许规避与胚胎杀伤性或缺乏特定的CRE驱动小鼠相关的潜在问题。这项技术的另一个相关方面是，它有助于减少涉及产生新的小鼠线的经济和时间限制，这在研究不同发育年龄时起源于大脑遥远区域的细胞类型之间的相互作用方面变得尤为重要。在这里，我们描述了一个双电镀策略，能够瞄准空间和时间分离的细胞群。通过这种方法，我们可以用选定的荧光蛋白标记不同位置的不同亚型细胞，以可视化它们，和/或者我们可以在适当的时候操纵这些不同细胞表达感兴趣的基因。这一策略增强了子宫电穿孔的潜力，并提供了一个强大的工具，用于研究迁移以建立密切接触的时空和空间分离的细胞群的行为，以及通过xxon投影进行远程相互作用，从而降低时间和经济成本。

相邻细胞之间的相互作用起源于远端位置和不同时间：卡贾尔-Retzius细胞（CR细胞）和早期迁移皮质投影神经元（策略A）
CR细胞和早期皮质投影神经元的相互作用先前被描述为有必要通过nectin和地籍粘附分子使用双电穿孔策略8来调节肌质易位。CR细胞起源于皮质边缘的神经上皮，并切向迁移，以填充皮层最表面的部分，边缘区域17，18，19，而皮?...

Watch this Scientific Journal Video about Double In Utero Electroporation to Target Temporally and Spatially Separated Cell Populations at JoVE.com

Double In Utero Electroporation to Target Temporally and Spatially Separated Cell Populations

子宫电穿孔的双倍允许瞄准空间和时间分离的细胞群。该技术可用于在正常条件下使用荧光蛋白可视化这些细胞群之间的相互作用，而且在功能实验后也可用于干扰感兴趣的基因。

双电穿孔，以定时和空间分离的细胞群

In utero electroporation is an in vivo DNA transfer technique extensively used to study the molecular and cellular mechanisms underlying mammalian ...

双子宫电穿孔是研究哺乳动物皮质生成的重要方面，如神经元细胞之间的相互作用，在不同位置和发育年龄产生的有力方法。这种技术的主要优点是能够以快速、详细和廉价的方式检查这些不同细胞群之间的不受干扰的相互作用。我们方法的一个显著含义是，它有助于更好地了解我们的接线在孤独症和精神分裂症等神经元疾病中是如何变化的。这项研究不仅限于新皮质，但它也可用于分析细胞与细胞的相互作用，在外皮质区域，如囊性或小脑。最具挑战性的步骤之一是将DNA注射到早期胚胎的横向心室。使用明亮的光源对于成功注射至关重要。对于每个手术，首先准备一个10微升溶液，其中包含一微升的快速绿色染料，每微升质粒DNA溶液1微克，每个质粒感兴趣的，以及适当的体积内毒素无TE缓冲液。并预装玻璃微胶囊移液器与适当准备的DNA溶液。对于子宫电穿孔手术中的第一个，在适当时间麻醉的怀孕小鼠的眼睛上涂抹软膏，并使用剃须刀剃光腹部。在腹部右侧进行切口之前，使用连续的70%乙醇和碘湿巾对裸露的皮肤进行两次消毒。一旦子宫被访问使用环钳仔细提取器官。使用口腔控制的吸气管，将大约0.5微升溶液注射到E11.5胚胎左半球的侧心室，用于策略 A.或注射到E13.5胚胎右半球的侧心室，用于策略B，直到心室内可见快速的绿色染料。当所有溶液都注射后，将钳子在注射胚胎头部的侧向周围放置钳子，将正极定向到中壁，以定位策略 A 的皮质下摆，或定向到策略 B 的侧皮层，注意避开心脏和胎盘。使用方波电波电波器为每个胚胎年龄应用适当的电脉冲序列，如表中所示。当所有胚胎都经过注射和电穿孔后，使用钳子小心地将子宫返回到腹腔，并在腹部充满温暖的盐水。使用针6-0缝合，关闭腹壁，并小心关闭皮肤切口。然后将动物返回到其家庭笼子进行监测，直到完全恢复。在最初的手术两天后，确认小鼠表现出正常行为，并且没有疼痛或不适的迹象。在准备动物的第二个子宫电穿孔之前， 刚才证明。将所需DNA溶液的0.5微升注入E13.5胚胎的左侧脑心室，用于策略B;每次注射后，将正极的电极向注射半球的侧皮层放置，用于策略B和策略B。以及表所示的电相胚胎。电穿孔完成后，将子宫返回到腹腔，并完成手术和手术后护理，如证明。在实施策略 A 时，在第二次电穿孔后四天，将胚胎日17.5怀孕雌性小鼠的子宫收获成冰上 PBS 的培养皿。在解剖显微镜下，用钳子将胚胎移植到PBS的新盘中。使用钳子，小心地抓住每个胚胎的头部，以方便去除头部和头骨上的皮肤。使用钳子或铲子将暴露的大脑抬出。然后用勺子将每个大脑转移到一个48孔板的单个孔中，每个井含有大约600微升的固定溶液。当所有的大脑被收集起来后，在轨道摇床上一夜之间将大脑固定在摄氏四度，然后用PBS进行三次十分钟的洗涤。上次洗涤后，将大脑嵌入4%的低熔点，在PBS中约10分钟。凝固后，使用氰丙烯酸酯胶将每个大脑固定到振动组织支架嗅球侧向上。将支架放在振动容器内，然后用 PBS 填充容器。然后使用振动器和刷子获得 100 微米厚的日冕序列段。每孔最多转移7个部分，每个胚胎6个孔，在一个新的48孔板中，每孔含有600微升的新鲜PBS，在收集时辅以抗真菌防腐剂。获得所有部分后，使用细毛刷将每个截面安装到玻璃显微镜幻灯片上，并用玻璃盖玻片盖住它们，以评估直立荧光显微镜上的电穿孔效果。在实施策略B时，在产后第15天，在确认对针头缺乏反应后，将鼠标固定在上针位置，并使用剪刀制作中线皮肤切口，露出肋骨和隔膜，并切开隔膜以进入心脏。使用精细剪刀，在右心房内做一个切口，并使用连接到柔性管的25个精袋针到渗透灌注泵穿透左心室。当针头就位时，通过转心灌注，以每分钟5.5毫升的恒定通量，提供至少25毫升4%的对甲醛。大约五分钟后，停止灌注，并使用剪刀去除头部的皮肤。取出头骨，用铲子取出脑组织。将大脑放入含有1.5毫升新鲜4%甲醛的24井板的一个井中，在轨道摇床上4摄氏度的夜间孵育。然后获得40微米的产后大脑部分，正如刚刚证明的。对于胚胎和产后脑片成像，将包含感兴趣的安装大脑部分的幻灯片放在显微镜幻灯片支架上。在共体显微镜中引入滑块支架，并选择实验中使用的氟化物的适当通道。执行样本扫描，以获取两种不同波长的每个大脑部分的地图图像，以便对双电穿孔输出进行一般查看。完成后，选择 10 倍镜头，在多区域 Z 堆栈时间差观察模式下。在每个选择的 XY 区域中，根据样品的平面设置适当的成像周长以及沿 Z 轴的扫描深度，其中荧光可见。获取所有选定区域的低放大倍率图像，并在显微镜查看器软件中将图像从 OIF 导出为 TIFF 格式。然后选择 60 倍镜头，并像刚才演示的一样捕获高放大倍率图像，以更详细的方式观察细胞到细胞的交互。子宫电穿孔手术的双重时间差距使CAG在胚胎第11.5天的目标细胞到达横向新皮质的边际区域，包括躯体感觉区，及时与皮质投影神经元建立接触，标记在胚胎日13.5。投影神经元表达增强的GP在皮层边缘区域大量上升，并与M-cherry表达猫部细胞的过程进行交联。在胚胎第13.5天的子宫电穿孔中，使用BFP表达质粒有助于不同皮质层投影神经元的标签。胚胎第15.5天反侧的第二次电穿孔针对的是上层细胞投影神经元亚群，而不是早期发育的下层投影神经元。上层目标神经元将其轴突扩展到反边半球，具有特征的轴化模式。高放大倍率允许更详细地可视化反边半球内目标投影神经元的角轴体内化。在尝试此协议时，必须根据要定位的细胞群规划电穿孔时间、注射半球以及电极的位置。遵循这个程序的所有方法，如电穿孔是有趣的一个。对于两种细胞类型，可以执行研究体内可能的电路变化。使用这种技术，可以研究两个不同的靶细胞之间的精细相互作用。包括体内新的科学信息，通过荧光突触蛋白的电穿孔。

Research

JoVE Journal

Neuroscience

Ultrasound-Guided Microinjection into the Mouse Forebrain In Utero at E9.5

Ultrasound-Guided Microinjection into the Mouse Forebrain In Utero at E9.5

超声引导下显微注射到小鼠前脑在子宫内在E9.5

In utero survival surgery in mice permits the molecular manipulation of gene expression during development.  However, because the uterine wall is opaque ...

科研

神经科学

In utero Electroporation followed by Primary Neuronal Culture for Studying Gene Function in Subset of Cortical Neurons

In utero Electroporation followed by Primary Neuronal Culture for Studying Gene Function in Subset of Cortical Neurons

在子宫内的电穿孔其次原发性神经文化为研究基因的功能在皮层神经元的一个子集

In vitro study of primary neuronal cultures allows for quantitative analyses of neurite outgrowth. In order to study how genetic alterations affect ...

Efficient Gene Delivery into Multiple CNS Territories Using In Utero Electroporation

Efficient Gene Delivery into Multiple CNS Territories Using In Utero Electroporation

高效的基因传递到多个中枢神经系统使用新界在子宫内电穿孔

The ability to manipulate gene expression is the cornerstone of modern day experimental embryology, leading to the elucidation of multiple developmental ...

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

在子宫内电穿孔：鼠标控制时空基因Transefection

In order to understand the function of genes expressed in specific region of the developing brain, including signaling molecules and axon guidance ...

Expansion of Embryonic and Adult Neural Stem Cells by In Utero Electroporation or Viral Stereotaxic Injection

Expansion of Embryonic and Adult Neural Stem Cells by In Utero Electroporation or Viral Stereotaxic Injection

扩展胚胎及成体神经干细胞<em&gt;在子宫内</em电穿孔或病毒立体定向注射

Somatic stem cells can divide to generate additional stem cells (expansion) or more differentiated cell types (differentiation), which is fundamental for ...

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

可视化和遗传操作在小鼠大脑皮层和使用海马树突和棘<em&gt;在子宫内</em&gt;电穿孔

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date ...

Generation of Topically Transgenic Rats by In utero Electroporation and In vivo Bioluminescence Screening

Generation of Topically Transgenic Rats by In utero Electroporation and In vivo Bioluminescence Screening

外用转基因大鼠通过代<em&gt;在子宫内</em&gt;电穿孔和<em&gt;在体内</em&gt;生物发光筛选

 In utero electroporation (IUE) is a technique which allows genetic modification of cells in the brain for investigating neuronal development. So far, the ...

Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation

Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation

遗传操纵鼠标的发展下丘脑通过<em&gt;在子宫内</em&gt;电穿孔

Genetic  modification of specific regions of the developing mammalian brain is a very  powerful experimental approach. However, generating novel mouse ...

Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse

Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse

Cortex-，Hippocampus-，Thalamus-，下丘脑 - ，横向隔核 - 纹状体专用<em&gt;在子宫内</em&gt;电穿孔的C57BL / 6小鼠

In utero electroporation is a widely used technique for fast and efficient spatiotemporal manipulation of various genes in the rodent central nervous ...

A Novel Strategy Combining Array-CGH, Whole-exome Sequencing and In Utero Electroporation in Rodents to Identify Causative Genes for Brain Malformations

A Novel Strategy Combining Array-CGH, Whole-exome Sequencing and In Utero Electroporation in Rodents to Identify Causative Genes for Brain Malformations

一种新的结合阵列全息、全外测序和宫内电穿孔的方法来识别脑畸形的致病基因

Birth defects that involve the cerebral cortex &#8212; also known as malformations of cortical development (MCD) &#8212; are important causes of ...