Name: investigating mammalian axon regeneration: in vivo electroporation of adult mouse dorsal root ganglion
Uploaded: 2018-09-01T14:30:00
Description: Watch this Scientific Journal Video about Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion at JoVE.com

这项研究获得资助 (授予 F Q。Z.) 由 NIH (R01NS064288, R01NS085176, R01GM111514, R01EY027347), 克雷格 h 尼尔森基金会和 BrightFocus 基金会。

所有动物实验都是按照约翰霍普金斯机构动物保育和使用委员会批准的动物协议进行的。
1. 材料和试剂
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<li>用六周大的雌性 CF1 小鼠称量 30–35 g 进行实验。 注: 在单独通风灭菌笼中, 小鼠组内 (每笼5只小鼠), 1/4 "玉米芯寝具和 2" 方形 nestlets 用于筑巢。笼保持在一个控制的12小时的光暗循环和室温是介于19.4 摄氏度和25摄氏度之间。用全球啮齿动...

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	<li>Saijilafu, , 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=PI3K-GSK3+signalling+regulates+mammalian+axon+regeneration+by+inducing+the+expression+of+Smad1.">PI3K-GSK3 signalling regulates mammalian axon regeneration by inducing the expression of Smad1.</a> Nature Communications. 4, 2690 (2013).</li><li>Zhang, B. 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=Akt-independent+GSK3+inactivation+downstream+of+PI3K+signaling+regulates+mammalian+axon+regeneration.">Akt-independent GSK3 inactivation downstream of PI3K signaling regulates mammalian axon regeneration.</a> Biochemical and Biophysical Research Communications. 443 (2), 743-748 (2014).</li><li>Jiang, J. 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=MicroRNA-26a+supports+mammalian+axon+regeneration+in+vivo+by+suppressing+GSK3beta+expression.">MicroRNA-26a supports mammalian axon regeneration in vivo by suppressing GSK3beta expression.</a> Cell Death &amp; Disease. 6, 1865 (2015).</li><li>Krames, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+dorsal+root+ganglion+in+the+development+of+neuropathic+pain.">The role of the dorsal root ganglion in the development of neuropathic pain.</a> Pain Medicine. 15 (10), 1669-1685 (2014).</li><li>Salimzadeh, L., Jaberipour, M., Hosseini, A., Ghaderi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-viral+transfection+methods+optimized+for+gene+delivery+to+a+lung+cancer+cell+line.">Non-viral transfection methods optimized for gene delivery to a lung cancer cell line.</a> Avicenna Journal of Medical Biotechnology. 5 (2), 68-77 (2013).</li><li>Keeler, A. M., ElMallah, M. K., Flotte, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+Therapy+2017:+Progress+and+Future+Directions.">Gene Therapy 2017: Progress and Future Directions.</a> Clinical and Translational Science. 10 (4), 242-248 (2017).</li><li>Neumann, E., Schaeferridder, M., Wang, Y., Hofschneider, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene-Transfer+into+Mouse+Lyoma+Cells+by+Electroporation+in+High+Electric-Fields.">Gene-Transfer into Mouse Lyoma Cells by Electroporation in High Electric-Fields.</a> The EMBO Journal. 1 (7), 841-845 (1982).</li><li>Liu, C. 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=MicroRNA-138+and+SIRT1+form+a+mutual+negative+feedback+loop+to+regulate+mammalian+axon+regeneration.">MicroRNA-138 and SIRT1 form a mutual negative feedback loop to regulate mammalian axon regeneration.</a> Genes &amp; Development. 27 (13), 1473-1483 (2013).</li><li>Saito, T., Nakatsuji, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+gene+transfer+into+the+embryonic+mouse+brain+using+in+vivo+electroporation.">Efficient gene transfer into the embryonic mouse brain using in vivo electroporation.</a> Developmental Biology. 240 (1), 237-246 (2001).</li><li>Saijilafu, E. M., Hur, F. Q., Zhou, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+dissection+of+axon+regeneration+via+in+vivo+electroporation+of+adult+mouse+sensory+neurons.">Genetic dissection of axon regeneration via in vivo electroporation of adult mouse sensory neurons.</a> Nature Communications. 2, 543 (2011).</li><li>Pan, 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=Shrinkage-mediated+imaging+of+entire+organs+and+organisms+using+uDISCO.">Shrinkage-mediated imaging of entire organs and organisms using uDISCO.</a> Nature Methods. 13, 859 (2016).</li><li>Rao, R. D., Bagaria, V. B., Cooley, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Posterolateral+intertransverse+lumbar+fusion+in+a+mouse+model:+surgical+anatomy+and+operative+technique.">Posterolateral intertransverse lumbar fusion in a mouse model: surgical anatomy and operative technique.</a> Spine Journal. 7 (1), 61-67 (2007).</li><li>Dommisse, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+blood+supply+of+the+spinal+cord.+A+critical+vascular+zone+in+spinal+surgery.">The blood supply of the spinal cord. A critical vascular zone in spinal surgery.</a> The Journal of Bone and Joint Surgery. British Volume. 56 (2), 225-235 (1974).</li><li>Sorensen, D. R., Leirdal, M., Sioud, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+Silencing+by+Systemic+Delivery+of+Synthetic+siRNAs+in+Adult+Mice.">Gene Silencing by Systemic Delivery of Synthetic siRNAs in Adult Mice.</a> Journal of Molecular Biology. 327 (4), 761-766 (2003).</li></ol>

一些手术步骤需要特别注意。L4 和 L5 DRGs (somas 的位置), 控制坐骨神经, 需要正确识别和注射的基因结构。否则, 在坐骨神经轴突中, GFP 标记将消失。髂嵴可以被视为有用的解剖标志, 以确定 L4 和 L5 DRGs。在大多数小鼠中, L5 和 L6 椎骨之间的关节与髂嵴12相近。另外, L3 根背根可以选择代替 L5, 特别是对免疫组化或西方印迹13的化验, 因为 L5 背节的手术暴露通常会遇?...

神经损伤或各种神经退行性疾病引起的神经系统损伤通常导致运动、感官和认知功能的缺陷。最近, 大量的努力已经致力于再生效能重建的成人神经元, 以恢复生理 functionsof 受损神经元1,2,3。神经节中的感觉神经元是一群神经细胞, 它传达不同的感官刺激, 如疼痛, 体温, 触觉, 或身体姿势, 对大脑。这些神经元都是伪单极, 包含一个单轴突, 分叉一个分支向周围延伸, 另一个分支朝向脊髓4。DRGs 中的成年感觉神经元是少数成熟的哺乳动物神经元, 在受伤后能积极地再生它们的轴突。因此, 感觉轴突损伤作为研究脑轴突再生机制的重要模型被广泛应用. 
在体内基因转染技术的建立和灵活性比使用转基因动物的时间要少, 在研究神经系统的基因功能和信号通路方面起着至关重要的作用。主要技术可以分为两种方法: 基于仪器的和基于病毒的5。以病毒为基础的成体神经元体内基因传递可以提供精确的时空操纵基因表达6。然而, 劳动密集型过程涉及基于病毒的方法, 如生产和纯化含有所需基因的病毒微粒。此外, 许多病毒载体可以激活宿主的免疫系统, 这可能会干扰数据的采集、数据分析, 并可能误导实验结果的解释。电穿孔是一种典型的以仪器为基础的转染方法, 使用电子脉冲增加细胞和核膜的通透性, 这有利于基因载体或小 RNA 寡核苷酸从细胞外的空间流入7.体外电穿孔被广泛认为是一种瞬态但高效的策略, 用于操纵多种细胞类型的靶向基因表达。虽然在体内电穿孔只会导致短暂的基因表达与病毒载体相比, 转染效率低, 它具有不同的病毒方法的优势。例如, 它可以应用到几乎所有的组织和细胞7,8,9。此外, 对某些转录蛋白编码的基因或小 RNA 寡核苷酸 (例如, siRNAs, microRNAs) 的质粒可以直接注入靶组织, 然后电脉冲, 这使得程序减少了劳动, 并耗时。此外, 转染多质粒和 RNA 寡核苷酸同时进行单电穿孔是可能的。
我们建立了体内电穿孔的方法来操纵成年小鼠感觉神经元中的基因表达, 并成功地应用和验证这种方法在许多先驱研究1,2,3 ,8,10。在这里, 我们提出了一个详细的协议, 以促进使用这种方法, 未来的研究哺乳动物轴突再生。

<table border="0" cellpadding="0" cellspacing="0" style="width:1039px;" width="1037">
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	<tbody>
		<tr height="26">
			<td height="26" style="height:27px;width:335px;">ECM 830 Square wave electroporation system</td>
			<td style="width:232px;">BTX Harvard Apparatus</td>
			<td style="width:121px;">45-0052</td>
			<td style="width:351px;">For in vivo electroporation</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:335px;">Tweezertrodes electrodes</td>
			<td style="width:232px;">BTX Harvard Apparatus</td>
			<td style="width:121px;">45-0524</td>
			<td>For in vivo electroporation, 1 mm flat</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Picospritzer III</td>
			<td style="width:232px;">Parker Instrumentation</td>
			<td style="width:121px;">1096</td>
			<td>Intracellular Microinjection Dispense Systems</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Glass Capillary Puller</td>
			<td style="width:232px;">NARISHIGE</td>
			<td>PC-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Borosilicate Glass Capillaries</td>
			<td>World Precision Instruments, Inc.</td>
			<td style="width:121px;">1902328</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Stereo Dissection Microscope</td>
			<td style="width:232px;">Leica</td>
			<td style="width:121px;">M80</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Microsurgery Rongeur</td>
			<td style="width:232px;">F.S.T</td>
			<td style="width:121px;">16221-14</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Microsurgery Forceps</td>
			<td style="width:232px;">FST by DUMONT, Switzerland</td>
			<td style="width:121px;">11255-20</td>
			<td>Only for sciatic nerve crush</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Glass Capillary</td>
			<td style="width:232px;">World Precision Instruments, Inc.</td>
			<td style="width:121px;">TW100-4</td>
			<td>10 cm, standard wall</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Tape</td>
			<td style="width:232px;">Fisherbrand</td>
			<td style="width:121px;">15-901-30</td>
			<td>For fixing the mouse on the corkboard</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2, 2, 2-Tribromoethanol (Avertin)</td>
			<td style="width:232px;">Sigma-Aldrich</td>
			<td style="width:121px;">T48402</td>
			<td>Avertin stock solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-methyl-2-butanol</td>
			<td style="width:232px;">Sigma-Aldrich</td>
			<td style="width:121px;">152463</td>
			<td>Avertin stock solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">siRNA Fluorescent Universal Negative Control #1</td>
			<td style="width:232px;">Sigma-Aldrich</td>
			<td style="width:121px;">SIC003</td>
			<td>Non-target siRNA with fluorescence</td>
		</tr>
		<tr height="28">
			<td height="28" style="height:28px;width:335px;">microRNA Mimic Transfection Control with Dy547</td>
			<td style="width:232px;">Dharmacon</td>
			<td style="width:121px;">CP-004500-01-05</td>
			<td>Non-target microRNA with fluorescence</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Plasmids preparation kit</td>
			<td style="width:232px;">Invitrogen Purelink</td>
			<td style="width:121px;">K210016</td>
			<td>GFP-coding plasmid preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:335px;">Fast Green Dye</td>
			<td style="width:232px;">Millipore-Sigma</td>
			<td style="width:121px;">F7252</td>
			<td>For better visualization of the DRG outline during injection</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:335px;">Ketamine</td>
			<td style="width:232px;">Putney, Inc</td>
			<td style="width:121px;">NDC 26637-731-51</td>
			<td>Anesthesia induction</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:335px;">Xylazine</td>
			<td style="width:232px;">AnaSed</td>
			<td style="width:121px;">NDC 59399-110-20</td>
			<td>Anesthesia induction</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:335px;">Acetaminophen</td>
			<td style="width:232px;">McNeil Consumer Healthcare</td>
			<td style="width:121px;">NDC 50580-449-36</td>
			<td>Post-surgical pain relief</td>
		</tr>
	</tbody>
</table>

investigating mammalian axon regeneration: in vivo electroporation of adult mouse dorsal root ganglion

电穿孔是一种重要的非病毒基因转染方法, 将 DNA 质粒或小 RNA 分子引入细胞。背根神经节 (DRGs) 中的感觉神经元延伸一个单轴突, 两个分支。一个分支转到外围神经 (外围分支), 另一个分支进入脊髓通过背根 (中央分支)。神经损伤后, 外围分支恢复强劲, 而中心分支不再生。随着再生能力的提高, 感觉轴突再生已被广泛应用于研究哺乳动物轴突再生在周围神经系统 (三七) 和中枢神经系统 (CNS) 中的模型系统。在这里, 我们描述了一个以前建立的方法协议来操纵成熟的感官神经元在体内通过电穿孔的基因表达。在转染质粒或小 RNA 寡核苷酸 (siRNAs 或 microRNAs) 的基础上, 该方法既允许损失和增益功能的实验, 以研究基因利益或 microRNAs 的作用, 调节轴突再生在体内。此外,体内基因表达的操纵在相对较短的时间内可以在空间上和世俗地控制。该模型系统为研究哺乳动物轴突再生在体内调节的分子机制提供了一个独特的工具。

为了量化当前协议的细胞毒性, 并验证体内根节电穿孔的转染率是否足够高, 我们注射并转荧光标记的 microRNA 或 siRNA L4 和 L5 DRGs。分离 DRGs 经低温切片和免疫组化处理 (图 1A B)。在估计注射和电穿孔后的细胞存活率时, L4 和 L5 的完整 DRGs 通过冷冻切片和免疫组化进行了采集和处理。与完整 DRGs 的神经元密度相比, 神经元密度与 Tuj1 ?...

Watch this Scientific Journal Video about Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion at JoVE.com

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

电穿孔是一种有效的方法, 将兴趣基因传递给细胞。本文应用这种方法对成年小鼠背根神经节的神经元进行了研究, 并对其在体内的轴突再生模型进行了描述。

哺乳动物轴突再生: 成年小鼠背根神经节体内电穿孔的研究

Electroporation is an essential non-viral gene transfection approach to introduce DNA plasmids or small RNA molecules into cells. A sensory neuron in the ...

This method provides an in-vivo gene transfection technique to manipulate gene expression in adult mouse sensory neurons for future studies of mammalian exo-regeneration. The main advantage of this technique is that it's less labor and time consuming, and can both overexpress and knock down the target genes simultaneously and separately. To begin this procedure, mark both sides of the iliac crest of an anesthetized mouse with a fine marker.Draw a line connecting two points of the iliac crest to facilitate the identification of L5 DRGs positions. Then, make a three centimeter incision along the midline of the lower back with micro scissors. Afterward, detach the paraspinous muscles, such as the musculus multifidus and the musculus longissimus lumborum, from the L3 to S1 spinous processes, and expose the fascia joint of the L4-5 and L5-6.Next, use a micro rongeur to remove the fascia joints of the L4-5 and L5-6. Subsequently, remove the left neural arch of L4 and L5, to expose the dorsal side of the DRGs. For DRG injection, load the DNA plasmids or RNA oligos into the glass capillary pipette.Then, insert the tip of the capillary glass pipette, carefully, into the DRG, and gradually inject one microliter solution of DNA plasmids or RNA oligos using the intracellular micro injection dispense system. For electroporation, pinch the target DRG with the electrodes gently, and apply five square electric pulses with the electroporation system. Subsequently, close the muscle, then the skin layers with nylon sutures.Two or three days after the DRG electroporation, anesthetize the mouse intraperitoneally, tape its limbs on the cork board, and make a one centimeter incision 0.5 centimeters to the left side along the midline. Next, cut the muscles, such as gluteus maximus muscle and piriformis muscle, longitudinally. Then, expose the segment of the sciatic nerve between the greater sciatic foramen, and the sciatic notch.Crush the nerve with microsurgery forceps for 12 seconds, and mark the crush site with a nylon epineural suture. Afterward, close the muscle and skin layers with nylon suture. After profusion, separate the DRGs together with the nerve roots and sciatic nerve, carefully, with micro scissors and micro forceps under the dissection microscope.Then, transfer the sciatic nerve directly in 4%PFA overnight at four degrees celsius. To image and measure the fluorescence labeled sensory axons under the dissection microscope, strip off the attached tissue and membrane on the fixed sciatic nerve with micro scissors and micro forceps, carefully. Then, wash the nerve with PVS three times.Next, place the sciatic nerve on a slide and keep it straight. Add 80 microliters of anti-fade solution around the nerve, then lay a cover slip on it. Flatten the whole mounted tissue with pressure.Afterward, place the flattened tissue under the inverted epifluorescence microscope, equipped with an accessory for mosaic acquisition and image processing. When measuring the length of regenerated axons, trace all identifiable fluorescently labeled axons in the sciatic nerve from the crush site to the distal axon ends. When applying the current, in-vivo electroporation method to study axonal regeneration, the sciatic nerve was flattened and imaged.Every regenerated axon with distinctive trajectory and recognizable distal axon end, can be traced from the crush site indicated by the suture knot. The white arrowhead, arrow, and star, indicate three distinctive axon ends, all of which extend from the crush site. Additionally, the DRGs were electroporated with EGFP plasmids, and the spinal cord was harvested.The EGFP labeled, dorsal column axons, in the spinal cord, were imaged and identified in both the longitudinal and sagittal projecting images. Here is a detailed view of axons in the dorsal column, and here is a detailed view of axons in the the dorsal root entry zone. This image shows the sagittal projection of the EGFP labeled axons within the dorsal column.Alternatively, the confocal images can be processed with microscopy visualization software to reconstruct a 3D image. While attempting this procedure, it's important to remember to position the L4 and the L5 DRGs correctly, and to make a knot on the dural membrane without impaling the sciatic nerve, while labeling the crush site with a suture knot. After its development, this technique pave the way for researchers in the field of axon regeneration to explore genes that are required for PNS, natural axon regeneration, or genes that can further promote regeneration in adult mouse in-vivo.

Research

JoVE Journal

Neuroscience

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

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 ...

科研

神经科学

Genetic Study of Axon Regeneration with Cultured Adult Dorsal Root Ganglion Neurons

培养成年的背根神经节神经元轴突再生的遗传研究

It is well known that mature neurons in the central nervous system (CNS) cannot regenerate their axons after injuries due to diminished intrinsic ability ...

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

在癌神经侵袭的体外模型：背根神经节型号

One way that solid tumors disseminate is through neural invasion. This route is well-known in cancers of the head and neck, prostate, and pancreas. These ...

An Approach to Enhance Alignment and Myelination of Dorsal Root Ganglion Neurons

一种方法来提高背根神经节的对齐和髓鞘

Axon regeneration is a chaotic process due largely to unorganized axon alignment. Therefore, in order for a sufficient number of regenerated axons to ...

Dorsal Root Ganglion Injection and Dorsal Root Crush Injury as a Model for Sensory Axon Regeneration

背根神经节内注射和背根挤压伤为感官轴突再生模型

Achieving axon regeneration after nervous system injury is a challenging task. As different parts of the central nervous system (CNS) differ from each ...

Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine

Laminotomy 对猪腰背根神经节通路和注射的治疗

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive ...

Analysis of Immune Cells in Single Sciatic Nerves and Dorsal Root Ganglion from a Single Mouse Using Flow Cytometry

应用流式细胞仪分析单侧坐骨神经和背根神经节的免疫细胞

Nerve-resident immune cells in the peripheral nervous system (PNS) are essential to maintaining neuronal integrity in a healthy nerve. The immune cells of ...

Rapid Isolation of Dorsal Root Ganglion Macrophages

多尾根结鱼巨噬细胞的快速隔离

There are growing interests to study the molecular and cellular interactions among immune cells and sensory neurons in the dorsal root ganglia after ...

In Situ Visualization of Axon Growth and Growth Cone Dynamics in Acute Ex Vivo Embryonic Brain Slice Cultures

In Situ Visualization of Axon Growth and Growth Cone Dynamics in Acute Ex Vivo Embryonic Brain Slice Cultures

原位 急性 离体 胚胎脑切片培养物中轴突生长和生长锥动力学的可视化

During neuronal development, axons navigate the cortical environment to reach their final destinations and establish synaptic connections. Growth cones ...

In Vitro Myelination of Peripheral Axons in a Coculture of Rat Dorsal Root Ganglion Explants and Schwann Cells

大鼠背根神经节外植体和雪旺细胞共培养中外周轴突的体外髓鞘形成

The process of myelination is essential to enable rapid and sufficient signal transduction in the nervous system. In the peripheral nervous system, ...