Name: in utero electroporation of multiaddressable genome-integrating color (magic) markers to individualize cortical mouse astrocytes
Uploaded: 2020-05-21T15:00:00
Description: Watch this Scientific Journal Video about In Utero Electroporation of Multiaddressable Genome-Integrating Color MAGIC Markers to Individualize Cortical Mouse Astrocytes at JoVE.com

我们感谢 S. Fouquet 以及蒙彼利埃视觉和神经科学协会 （MRI 和 RAM） 的成像和动物核心设施提供的技术援助。这项工作得到了法国雷吉翁·伊勒-德-法兰西基金会和.C癌症康复基金会以及巴黎-萨克莱大学（跨学科博士倡议）到洛杉矶的奖学金的支持，欧洲研究理事会（ERC-SG 336331， PI J. 瓦莱特） 到 E. h.， 由国家重建局根据合同 ANR-10-LABX-65 （实验室生命感官）， ANR-11-EQPX-0029 （Equipex 莫福镜 2）， ANR-10-INBS-04 （法国生物成像），由欧洲研究理事会（ERC-CoG 649117，PI J. Livet）和ATIP-Avenir项目（PI K.Loulier）的雷切·梅迪卡莱基金会（DBI20141231328）。

此处描述的所有动物程序都是按照机构准则进行的。动物协议已得到查尔斯·达尔文动物实验伦理委员会（CEEACD/N+5）的批准。
1. 子宫电镀中魔法标记物的无毒素质粒制备
<ol><li>细菌转化<ol>
<li>在冰上，解冻DH5阿尔法能力细胞存储在-70°C。</li>
		<li>在 37 °C 下加热含有适当抗生素（100μg/mL 氨基林或 50μg/mL 卡纳霉素）的琼脂板。</li>
		<li>在解冻的DH5α能力细胞中加入1 μL的5-50克...

<ol>
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在皮质祖细胞的魔法标记的子宫电冲（IUE）中（图1）使星形细胞在产后大脑皮层的标记在不同的产后阶段（P4-P7-P21，图2）。有趣的是，IUE 的阶段并不关键，因为从 E13.5 到 E15.5 执行的电冲产生与皮质星形细胞14类似的标记模式。然而，标记的金字塔神经元在皮质帕伦奇马的位置随电扑阶段而变化。事实上，IUE执行在E15标记层2-3神经元...

天体细胞在大脑发育和生理学中起着许多重要作用。除了在调节营养吸收和血液流动的血脑屏障中发挥作用外，他们还积极促进突触的形成和功能，同时产生神经调节器，可以改变神经元活动和行为2。此外，天体细胞功能障碍导致各种神经系统疾病3.位于大脑皮层的星形细胞显示一个精心设计的形态，能够广泛接触神经元过程。这些接触，对电路功能至关重要，也通过细胞粘附蛋白4控制星形细胞形态生成和突触发生。神经科学家需要方便和可靠的工具来研究天体细胞发育和形态生成在他们的神经模型的兴趣。然而，由于星形细胞与邻居的紧密对接和均匀的三维平铺，因此，挑出皮质星形细胞并使用免疫标志物全面评估其形态是具有挑战性的。
目前，两种主要的基因工程策略使皮质天体在原位进行标记和个性化：利用记者质粒的电聚化，在转基因小鼠系或体细胞变代中稀疏的记者激活。第一个策略依赖于繁殖一个飞毛腿记者小鼠线与小鼠表示一种诱导形式的Cre重组酶激活专门在天体细胞后塔莫西芬交付（如Aldh1l1-CreERT25）。此策略存在几个缺点。首先，繁殖转基因小鼠需要大量的动物，通常需要多次检测来确定适当剂量的塔莫西芬，以提供皮质星形细胞的足够稀疏标签。分析感兴趣的遗传小鼠模型中的皮质星体表型将需要更多的繁殖和小鼠消费。此外，在子宫塔莫西芬注射已知干扰分体，使这一策略难以应用于研究天体细胞发育的最初阶段。体内DNA电聚是一种替代的无塔莫西芬策略，它依赖于最少数量的动物6。这种方法在胚胎阶段或产后阶段进行，包括在啮齿动物的横向心室中注射报告质粒，然后用电脉冲在细胞膜中产生毛孔，从而允许DNA进入心室内的祖细胞。随后，记者携带的电聚合质粒的转基因由目标细胞机械处理，并表达7。此前曾描述过两种电冲洗方法，为小鼠皮质星形细胞贴上标签：1）产后星形细胞标签（PALE），它依赖于产后早期4-2个单色偶发性报告质粒的电冲洗：2）星轨策略基于在子宫电镀（IUE）的多色综合记者质粒8，9，10。虽然这两种技术在大脑皮层中有效地标记了PrA，但它们也存在一些局限性。在其初始版本中，这两种方法都依赖于胶质纤维酸性蛋白 （GFAP） 促进剂来驱动星形细胞中的表达，这可能将标签偏向径向胶质细胞，以及表达 GFAP 比正常休息 PrA11、12更强烈的皮亚和活性星形细胞。关于PALE，其他缺点是电冲洗的后期阶段，它防止标记早产的PrA（或那些源于早期脱脂的祖先）和分析天体发育的早期阶段，以及使用偶发性病媒，在PrA在第13、14周经历的大规模增殖期间通过连续的分裂被稀释。与 PALE 不同的是，StarTrack 基于集成记者质粒的胚胎电聚，允许跟踪胚胎和产后祖先对 PrA 的贡献。依靠泛素C促进剂（UbC-StarTrack）的更新星轨方案实现了荧光记者在神经祖祖15、16、17的神经元和胶质下降（包括星形细胞）的更广泛表达。然而，在其当前版本中，这种方法的实施是复杂的，因为它依赖于12种不同质粒的等效混合物，表达6种荧光蛋白（FP），部分激发和发射光谱重叠。
这里介绍的是一个直接的子宫电镀基多色标签方法使用综合记者结构驱动的强大和广泛活跃的促进者挑出皮质星形细胞14。此外，还分别提供了使用许可（如 Imaris）和开放访问（Vaa3D18、19、20）图像分析软件的简单图像分析管道，用于分割天体细胞的面积体积和树干。与之前描述的方法相比，此策略仅依赖于 1-2 个多色集成转基因多易处理基因组集成颜色标记（MAGIC 标记或 MM21），这些标记指向细胞质和（可选）核细胞隔间，其表达由由环状体增强剂组成的合成CAG促进器驱动， 鸡β蛋白推广器，兔β-红蛋白拼接接受网站22。这使得皮质星形细胞的标签和跟踪，从胚胎阶段到产后后期阶段，独立于GFAP表达14，23。这些转基因中的每一种都具有以下四种不同的FP：eBFP、绿松石2/mCerulean、EYFP和tdTomato/mCherry，它们显示最小的光谱重叠，可以很容易地绕过1）顺序通道采集：2） 优化激发能力和收集增益：和3）特定的二氯气过滤器，以收集狭窄的FP排放窗口。MM 策略使用Cre/lox重组与自切除的 Cre 重组 （seCre） 来驱动蜂窝群中 FP 的随机表达。MM转基因的单个副本以相互排斥的方式表达FP，而多个转基因则产生FP组合，产生几十种不同的色调。转基因的基因组整合是由小猪巴克（PB）或托尔2转位系统24，25，26驱动的。因此，通过子宫电冲，MM工具包和多色"马赛克"，它产生启用多个相邻的皮质祖先的同步标记和跟踪其胶质下降，包括皮质星形细胞，在很长一段时间内。颜色对比产生于对 PrA 轮廓的明显 FP 允许线的表达，随后提取有关其领土体积（使用 IMARIS）和复杂形态（使用 Vaa3D）的关键信息。这里详细介绍的多色策略是一种方便而可靠的方法，可在各种发育阶段快速、便捷地访问野生型小鼠的皮质星形细胞表面和形态，并且无需使用转基因记者线即可轻松适应研究神经系统疾病小鼠模型中的天体细胞解剖特征。

<table>
	<tbody>
		<tr>
			<td>1.1 Bacteria transformation</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ampicillin</td>
			<td>Euromedex</td>
			<td>EU0400-C</td>
			<td></td>
		</tr>
		<tr>
			<td>DH5 alpha competent cells</td>
			<td>Fisher Scientitic</td>
			<td>11563117</td>
			<td></td>
		</tr>
		<tr>
			<td>Ice box</td>
			<td>Dutscher</td>
			<td>139959</td>
			<td></td>
		</tr>
		<tr>
			<td>Kanamycin</td>
			<td>Sigma</td>
			<td>60615</td>
			<td></td>
		</tr>
		<tr>
			<td>LB Agar</td>
			<td>Sigma</td>
			<td>L2897</td>
			<td></td>
		</tr>
		<tr>
			<td>SOC medium</td>
			<td>Fisher Scientitic</td>
			<td>11563117</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile petri dish- 10 cm</td>
			<td>Thermo Fisher</td>
			<td>150350</td>
			<td></td>
		</tr>
		<tr>
			<td>Water bath</td>
			<td>VWR</td>
			<td>462-0556H</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1.2 Plasmid culture</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>14 ml culture tube</td>
			<td>Dutscher</td>
			<td>187262</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass erlenmeyer- 2L</td>
			<td>Fisher Scientitic</td>
			<td>11383454</td>
			<td></td>
		</tr>
		<tr>
			<td>LB medium</td>
			<td>Sigma</td>
			<td>L3522</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1.3 Plasmid DNA preparation</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NucleoBond Xtra Maxi Plus EF</td>
			<td>Macherey-Nagel</td>
			<td>740426.10</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2.1 Preparation of the solutions</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>26 G x 1/2 needle</td>
			<td>Terumo</td>
			<td>8AN2613R1</td>
			<td></td>
		</tr>
		<tr>
			<td>30 G x 1/2 needle</td>
			<td>Terumo</td>
			<td>8AN3013R1</td>
			<td></td>
		</tr>
		<tr>
			<td>Fast Green</td>
			<td>Sigma Aldrich</td>
			<td>F7272</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>VWR</td>
			<td>27810.295</td>
			<td></td>
		</tr>
		<tr>
			<td>Single-use polypropylene syringe, 1 mL</td>
			<td>Dutscher</td>
			<td>50002</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2.2 Preparation of the surgery material</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Adson Forceps - DeBakey Pattern- 12.5 cm</td>
			<td>FST</td>
			<td>11617-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Arched tip Forceps- 10 cm</td>
			<td>FST</td>
			<td>11071-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass bead sterilizer Steri 250</td>
			<td>Sigma</td>
			<td>Z378569</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass micropipette 1 mm diameter</td>
			<td>FHC</td>
			<td>10-10-L</td>
			<td></td>
		</tr>
		<tr>
			<td>Graefe Forceps - Titanium 1 mm Tips Slight Curve- 10 cm</td>
			<td>FST</td>
			<td>11651-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Graefe Forceps - Titanium 1 mm Tips Straight- 10 cm</td>
			<td>FST</td>
			<td>11650-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Iris Scissors - Delicate Straight- 9 cm</td>
			<td>FST</td>
			<td>14060-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Laboratory tape</td>
			<td>Fisher Scientitic</td>
			<td>11730454</td>
			<td></td>
		</tr>
		<tr>
			<td>Microinjector</td>
			<td>INJECT+MATIC</td>
			<td>No catalog number</td>
			<td></td>
		</tr>
		<tr>
			<td>Olsen-Hegar Needle Holder - 12 cm</td>
			<td>FST</td>
			<td>12002-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Optical fiber</td>
			<td>VWR</td>
			<td>631-1806</td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic-coated white paper</td>
			<td>Distrimed</td>
			<td>700103</td>
			<td></td>
		</tr>
		<tr>
			<td>Signagel electrode gel</td>
			<td>Free-Med</td>
			<td>15-60</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Petri dish- 35 mm</td>
			<td>Dutscher</td>
			<td>056714</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterilizer, glass dry bead, Steri 250</td>
			<td>Sigma</td>
			<td>Z378569</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2.3 Preparation of the pregnant female mouse</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol pad</td>
			<td>Alcomed</td>
			<td>1731000</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprecare</td>
			<td>Axience</td>
			<td>0.3 mg/ml</td>
			<td></td>
		</tr>
		<tr>
			<td>Compress</td>
			<td>tRAFFIN</td>
			<td>70189</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Merial</td>
			<td>Imalgene 1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Ocular gel</td>
			<td>tvm lab</td>
			<td>Ocry-gel</td>
			<td></td>
		</tr>
		<tr>
			<td>RjOrl:SWISS mice</td>
			<td>Janvier Labs</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vetadine, 10% solution</td>
			<td>Vetoquinol</td>
			<td>4337400113B</td>
			<td></td>
		</tr>
		<tr>
			<td>Warming pad</td>
			<td>Harvard Apparatus</td>
			<td>72-0493</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Bayer</td>
			<td>Rompun 2%</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>3.2 Electroporation</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbable suture Size 4-0 45 cm Suture 1-Needle 19 mm Length 3/8 Circle Reverse</td>
			<td>Novosyn</td>
			<td>C0068220</td>
			<td></td>
		</tr>
		<tr>
			<td>Electroporateur Sonidel</td>
			<td>Sonidel</td>
			<td>NEPA 21</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile transfer pipets (individually wrapped)</td>
			<td>Dutscher</td>
			<td>043202S</td>
			<td></td>
		</tr>
		<tr>
			<td>Tweezers with 3 mm platinium disk electrodes</td>
			<td>Sonidel</td>
			<td>CUY650P3</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>4.1 Tissue harvesting and sectioning</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>24-well plate</td>
			<td>Falcon</td>
			<td>353047</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>Lonza</td>
			<td>50004</td>
			<td></td>
		</tr>
		<tr>
			<td>Antigenfix</td>
			<td>Microm Microtech</td>
			<td>U/P0014</td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>Dutscher</td>
			<td>100266</td>
			<td></td>
		</tr>
		<tr>
			<td>Dolethal</td>
			<td>Vetoquinol</td>
			<td>DOL202</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS (10X), no calcium, no magnesium</td>
			<td>Fisher Scientific</td>
			<td>11540486</td>
			<td></td>
		</tr>
		<tr>
			<td>Nail polish</td>
			<td>EMS</td>
			<td>72180</td>
			<td></td>
		</tr>
		<tr>
			<td>Slide</td>
			<td>Dutscher</td>
			<td>100001</td>
			<td></td>
		</tr>
		<tr>
			<td>Vectashield</td>
			<td>Vectorlabs</td>
			<td>H-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Vibratome</td>
			<td>Leica</td>
			<td>VT1000S</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>5. Multichannel confocal imaging</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>20X oil NA 0.85</td>
			<td>Olympus</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Confocal Laser Scanning Microscope</td>
			<td>Carl Zeiss</td>
			<td>LSM880</td>
			<td></td>
		</tr>
		<tr>
			<td>Confocal Laser Scanning Microscope</td>
			<td>Olympus</td>
			<td>FV1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Plan Apochromat 20x/0.8 M27</td>
			<td>Carl Zeiss</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>6. Astrocyte territorial volume segmentation</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>IMARIS 8.3 and later versions</td>
			<td>Bitplane</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>7. Astrocyte arborization tracing</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>3D Visualization-Assisted Analysis software suite (Vaa3D)</td>
			<td>HHMI - Janelia Research Campus /Allen Institute for Brain Science</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

in utero electroporation of multiaddressable genome-integrating color (magic) markers to individualize cortical mouse astrocytes

位于小鼠大脑皮层的原质天体细胞（PrA）紧密并列，在成人阶段形成明显连续的三维矩阵。到目前为止，没有任何免疫控制策略可以挑出它们，并在成熟动物和皮质遗传过程中分割它们的形态。皮质PrA起源于位于鼻腔姑息的祖先，在综合载体的子宫电渗透中很容易被瞄准。这里提出了一个协议，为这些细胞贴上多地址基因组整合颜色（MAGIC）标记策略的标签，该策略依赖于小猪/Tol2的转换和Cre/lox 重组，以固态地表达特定的亚细胞隔间的不同荧光蛋白（蓝色、青色、黄色和红色）。这种多色命运映射策略能够在胶质发生之前用颜色标记的组合在附近的皮质祖先进行原位标记，并在单个细胞水平上跟踪其后代，包括星形细胞，从胚胎阶段到成人阶段。通过调整多地址基因组集成颜色标记（MAGIC 标记或 MM）提供的电冲积载体的浓度和颜色对比度实现的半稀疏标记，能够个性化星形细胞，并挑出其区域和复杂的形态，尽管它们具有密集的解剖排列。这里介绍的是一个全面的实验工作流程，包括电化过程的细节，多通道图像堆栈通过胸腔显微镜采集，以及计算机辅助的三维分割，使实验者能够评估单个PrA体积和形态。总之，MAGIC 标记器的电冲控提供了一种方便的方法，可以单独标记许多星形细胞，并在不同的发育阶段访问其解剖特征。这项技术将可用于分析各种小鼠模型中的皮质星形体形态特性，而无需使用具有转基因记者线的复杂十字架。

胚胎皮质祖细胞中魔法标记的电化允许从大脑皮层发育的早期到后期标记星形细胞（图1）。这些星形细胞在产后各阶段（P4、P7、P21）的所有皮质层中被发现，因为它们广泛分散到整个大脑皮层中。他们用以 20x 0.8 NA（或更高 NA）目标获得的瓷砖组合图像进行评估，并组装为 Z 堆栈重建（图 2）。MAGIC 标记组合标记启用皮质星形细胞的个性化，并提取有?...

Watch this Scientific Journal Video about In Utero Electroporation of Multiaddressable Genome-Integrating Color MAGIC Markers to Individualize Cortical Mouse Astrocytes at JoVE.com

In Utero Electroporation of Multiaddressable Genome-Integrating Color MAGIC Markers to Individualize Cortical Mouse Astrocytes

星形细胞均匀地平铺大脑皮层，使得对其复杂形态的分析在细胞水平上具有挑战性。此处提供的协议使用基于子宫电镀的多色标签，挑出皮质星形细胞，并使用用户友好的图像分析管道分析其体积和形态。

在多地址基因组集成颜色 （MAGIC） 标记的 Utero 电冲中， 以个性化皮质小鼠星形细胞

Protoplasmic astrocytes (PrA) located in the mouse cerebral cortex are tightly juxtaposed, forming an apparently continuous three-dimensional matrix at ...

Research

JoVE Journal

Neuroscience

In Utero Electroporation of Multiaddressable Genome-Integrating Color (MAGIC) Markers to Individualize Cortical Mouse Astrocytes

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 ...

科研

神经科学

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

小鼠视网膜神经节细胞的转染在体内电穿孔

The targeting and refinement of RGC projections to the midbrain is a popular and powerful model system for studying how precise patterns of neural ...

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 ...

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 ...

Isolation and Culture of Mouse Cortical Astrocytes

小鼠脑皮层星形胶质细胞的分离和培养

Astrocytes  are an abundant cell type in the mammalian brain, yet much remains to be  learned about their molecular and functional characteristics. In ...

Ex utero Electroporation and Whole Hemisphere Explants: A Simple Experimental Method for Studies of Early Cortical Development

Ex utero Electroporation and Whole Hemisphere Explants: A Simple Experimental Method for Studies of Early Cortical Development

前子宫穿孔和整个半球的外植体早期皮质发育的研究：一个简单的实验方法

Cortical development involves complex interactions between neurons and non-neuronal elements including precursor cells, blood vessels, meninges and ...

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 ...

Subtype-selective Electroporation of Cortical Interneurons

皮质的interneurons的亚型选择性电穿孔

The study of central nervous system (CNS) maturation relies on genetic targeting of neuronal populations. However, the task of restricting the expression ...

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 ...

在多地址基因组集成颜色 （MAGIC） 标记的 Utero 电冲中， 以个性化皮质小鼠星形细胞

在多地址基因组集成颜色（MAGIC）标记的 Utero 电冲中，以个性化皮质小鼠星形细胞