Name: rapid isolation of dorsal root ganglion macrophages
Uploaded: 2019-09-07T14:30:00
Description: Watch this Scientific Journal Video about Rapid Isolation of Dorsal Root Ganglion Macrophages at JoVE.com

这项研究得到了麻醉教育与研究基金会（XY）的支持;UCSF麻醉和围手术期护理部（XY）;和 1R01NS100801-01 （GZ）。这项研究部分得到了HDFCCC细胞分析共享资源实验室的一部分支持，得到了NIH（P30CA082103）的资助。

所有动物实验均获得加州大学旧金山分校机构动物护理和使用委员会的批准，并根据NIH《实验室动物护理和使用指南》进行。
1. 从实验鼠身上收集腰椎DRG
<ol><li>在开始实验之前，通过将 9 卷介质与 1 卷 Ca+/Mg=-10x HBSS 混合，准备密度梯度介质（例如 Percoll）的工作溶液。把它放在冰上</li>
	<li>用2.5%的Avertin麻醉小鼠。确认动物完全麻醉，由于缺乏对后爪捏的反?...

<ol>
	<li>Ji, R. R., Chamessian, A., Zhang, Y. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pain+regulation+by+non-neuronal+cells+and+inflammation.">Pain regulation by non-neuronal cells and inflammation.</a> Science. 354 (6312), 572-577 (2016).</li><li>Inoue, K., Tsuda, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microglia+in+neuropathic+pain:+cellular+and+molecular+mechanisms+and+therapeutic+potential.">Microglia in neuropathic pain: cellular and molecular mechanisms and therapeutic potential.</a> Nature Reviews Neurosciences. 19 (3), 138-152 (2018).</li><li>Hu, P., McLachlan, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+functional+types+of+macrophage+in+dorsal+root+ganglia+and+spinal+nerves+proximal+to+sciatic+and+spinal+nerve+transections+in+the+rat.">Distinct functional types of macrophage in dorsal root ganglia and spinal nerves proximal to sciatic and spinal nerve transections in the rat.</a> Experimental Neurology. 184 (2), 590-605 (2003).</li><li>Simeoli, R., Montague, K., Jones, H. 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=Exosomal+cargo+including+microRNA+regulates+sensory+neuron+to+macrophage+communication+after+nerve+trauma.">Exosomal cargo including microRNA regulates sensory neuron to macrophage communication after nerve trauma.</a> Nature Communication. 8 (1), 1778 (2017).</li><li>Cobos, E. J., Nickerson, C. A., Gao, 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=Mechanistic+differences+in+neuropathic+pain+modalities+revealed+by+correlating+behavior+with+global+expression+profiling.">Mechanistic differences in neuropathic pain modalities revealed by correlating behavior with global expression profiling.</a> Cell Reports. 22 (5), 1301-1312 (2018).</li><li>Zhang, H., Li, Y., de Carvalho-Barbosa, 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=Dorsal+Root+Ganglion+Infiltration+by+Macrophages+Contributes+to+Paclitaxel+Chemotherapy-Induced+Peripheral+Neuropathy.">Dorsal Root Ganglion Infiltration by Macrophages Contributes to Paclitaxel Chemotherapy-Induced Peripheral Neuropathy.</a> The Journal of Pain. 17 (7), 775-786 (2016).</li><li>Liu, T., van Rooijen, N., Tracey, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depletion+of+macrophages+reduces+axonal+degeneration+and+hyperalgesia+following+nerve+injury.">Depletion of macrophages reduces axonal degeneration and hyperalgesia following nerve injury.</a> Pain. 86 (1-2), 25-32 (2000).</li><li>Peng, J., Gu, N., Zhou, 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=Microglia+and+monocytes+synergistically+promote+the+transition+from+acute+to+chronic+pain+after+nerve+injury.">Microglia and monocytes synergistically promote the transition from acute to chronic pain after nerve injury.</a> Nature Communication. 7, 12029 (2016).</li><li>Barclay, J., Clark, A. K., Ganju, P., 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=Role+of+the+cysteine+protease+cathepsin+S+in+neuropathic+hyperalgesia.">Role of the cysteine protease cathepsin S in neuropathic hyperalgesia.</a> Pain. 130 (3), 225-234 (2007).</li><li>Lim, H., Lee, H., Noh, K., Lee, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IKK/NF-kappaB-dependent+satellite+glia+activation+induces+spinal+cord+microglia+activation+and+neuropathic+pain+after+nerve+injury.">IKK/NF-kappaB-dependent satellite glia activation induces spinal cord microglia activation and neuropathic pain after nerve injury.</a> Pain. 158 (9), 1666-1677 (2017).</li><li>Rutkowski, M. D., Pahl, J. L., Sweitzer, S., van Rooijen, N., DeLeo, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limited+role+of+macrophages+in+generation+of+nerve+injury-induced+mechanical+allodynia.">Limited role of macrophages in generation of nerve injury-induced mechanical allodynia.</a> Physiology and Behavior. (3-4), 225-235 (2000).</li><li>Kwon, M. J., Shin, H. Y., Cui, 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=CCL2+Mediates+Neuron-Macrophage+Interactions+to+Drive+Proregenerative+Macrophage+Activation+Following+Preconditioning+Injury.">CCL2 Mediates Neuron-Macrophage Interactions to Drive Proregenerative Macrophage Activation Following Preconditioning Injury.</a> Journal of Neuroscience. 35 (48), 15934-15947 (2015).</li><li>Zigmond, R. E., Echevarria, F. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Macrophage+biology+in+the+peripheral+nervous+system+after+injury.">Macrophage biology in the peripheral nervous system after injury.</a> Progress in Neurobiology. 173, 102-121 (2019).</li><li>Ghasemlou, N., Chiu, I. M., Julien, J. P., Woolf, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD11b+Ly6G-+myeloid+cells+mediate+mechanical+inflammatory+pain+hypersensitivity.">CD11b+Ly6G- myeloid cells mediate mechanical inflammatory pain hypersensitivity.</a> Proceedings of the National Academy of Science USA. 112 (49), E6808-E6817 (2015).</li><li>Malin, S. A., Davis, B. M., Molliver, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+of+dissociated+sensory+neuron+cultures+and+considerations+for+their+use+in+studying+neuronal+function+and+plasticity.">Production of dissociated sensory neuron cultures and considerations for their use in studying neuronal function and plasticity.</a> Nature Protocols. 2 (1), 152-160 (2007).</li><li>Lin, Y. T., Chen, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dorsal+Root+Ganglia+Isolation+and+Primary+Culture+to+Study+Neurotransmitter+Release.">Dorsal Root Ganglia Isolation and Primary Culture to Study Neurotransmitter Release.</a> Journal of Visualized Experiment. (140), (2018).</li><li>Burnett, S. H., Kershen, E. J., Zhang, 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=Conditional+macrophage+ablation+in+transgenic+mice+expressing+a+Fas-based+suicide+gene.">Conditional macrophage ablation in transgenic mice expressing a Fas-based suicide gene.</a> J Leukoc Biol. 75 (4), 612-623 (2004).</li><li>Lopes, D. M., Malek, N., Edye, 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=Sex+differences+in+peripheral+not+central+immune+responses+to+pain-inducing+injury.">Sex differences in peripheral not central immune responses to pain-inducing injury.</a> Science Reports. 7 (1), 16460 (2017).</li><li>Kim, Y. S., Anderson, M., Park, 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=Coupled+Activation+of+Primary+Sensory+Neurons+Contributes+to+Chronic+Pain.">Coupled Activation of Primary Sensory Neurons Contributes to Chronic Pain.</a> Neuron. 91 (5), 1085-1096 (2016).</li><li>Vicuna, L., Strochlic, D. E., Latremoliere, A., 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+serine+protease+inhibitor+SerpinA3N+attenuates+neuropathic+pain+by+inhibiting+T+cell-derived+leukocyte+elastase.">The serine protease inhibitor SerpinA3N attenuates neuropathic pain by inhibiting T cell-derived leukocyte elastase.</a> Nature Medicine. 21 (5), 518-523 (2015).</li></ol>

在这里，我们介绍了一种新方法，有效地丰富了从小鼠DRG分离的巨噬细胞。分离DRG免疫细胞的传统方法需要酶消化15，18，现在在我们的协议中用机械均质代替，以限制不需要的细胞损伤，提高产量。因此，新协议省时少得多。更重要的是，酶消化可以刺激巨噬细胞并改变分子特征。相比之下，我们的机械方法极大地限制了细胞压力。
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现在有相当多的证据表明，免疫细胞在外周神经损伤1，2后造成神经病变疼痛。外周单细胞，包括成熟的巨噬细胞，已知通过噬菌体、抗原表达和细胞因子释放对组织损伤和全身感染作出反应。平行神经损伤引起的微胶质激活在脊髓背角，巨噬细胞在背根神经神经损伤（DRG）后也显著扩大后神经损伤3，4。值得注意的是，有越来越多的兴趣，以确定宏噬细胞是否有助于神经病变后，周围神经损伤后，与感觉神经元在DRG 5，6，7相互作用8,9,10,11.此外，最近的研究也涉及DRG巨噬细胞在神经损伤12、13后对斧道修复的贡献。另一项研究进一步表明，巨噬细胞亚群（即CD11b+Ly6CHi和CD11b+Ly6C低/-细胞）在机械超敏14中可能扮演不同的角色。因此，在神经损伤的情况下快速对DRG巨噬细胞的反应进行表示，可以帮助我们识别导致神经病痛的神经免疫因素。
按照惯例，在DRG中分离巨噬细胞的协议涉及多个步骤，包括酶消化15，16。该技术通常非常耗时，对于大规模实验来说可能成本高昂。虽然温和的消化与胶原酶II型（4毫克/mL）和消血酶II型（4.7毫克/mL）20分钟建议之前15，可以想象，细胞后暴露于这种酶是容易细胞损坏或细胞死亡，这可能导致低产量。此外，酶质量的差异可能进一步影响这一过程的效率。更重要的是，暴露于酶消化的巨噬细胞可能被无情地刺激，因此可能与体内状态非常不同。这些变化可能使功能性研究的结果复杂化。
在这里，我们描述了一种无酶协议，利用机械解散在4°C下快速分离DRG巨噬细胞。样品保存在冰上，以限制细胞压力。因此，我们的方法提供了保持隔离一致性的优势，并且分离的细胞可能更健康，刺激更少。我们进一步提出证据，通过荧光激活细胞分拣（FACS）分析验证分离细胞的质量。

<table>
	<tbody>
		<tr>
			<td>AP20187</td>
			<td>Clontech</td>
			<td>635058</td>
			<td></td>
		</tr>
		<tr>
			<td>a-mouse CX3CR1-APC antibody</td>
			<td>Biolegend</td>
			<td>149007</td>
			<td></td>
		</tr>
		<tr>
			<td>Avertin</td>
			<td>Sigma</td>
			<td>T48402</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell strainer (70 mm nylon)</td>
			<td>Falcon</td>
			<td>352350</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Eppendorf</td>
			<td>5810R</td>
			<td></td>
		</tr>
		<tr>
			<td>Dounce tissue homogenizer</td>
			<td>Wheaton</td>
			<td>357538 (1ml)</td>
			<td></td>
		</tr>
		<tr>
			<td>FACS tubes (5ml)</td>
			<td>Falcon</td>
			<td>352052</td>
			<td></td>
		</tr>
		<tr>
			<td>Friedman-Pearson Rongeur</td>
			<td>FST</td>
			<td>16121-14</td>
			<td></td>
		</tr>
		<tr>
			<td>HBSS (10x, Ca++/Mg++-free)</td>
			<td>Gibco</td>
			<td>14185-052</td>
			<td></td>
		</tr>
		<tr>
			<td>Noyes Spring Scissor</td>
			<td>FST</td>
			<td>15012-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Percoll</td>
			<td>Sigma</td>
			<td>P4937-500ml</td>
			<td></td>
		</tr>
		<tr>
			<td>Propidium iodide</td>
			<td>Sigma</td>
			<td>P4864-10ml</td>
			<td></td>
		</tr>
	</tbody>
</table>

rapid isolation of dorsal root ganglion macrophages

研究周围神经损伤后背根神经痛的背根神经瘤的免疫细胞和感觉神经元之间的分子和细胞相互作用的兴趣越来越大。外周单细胞，包括巨噬细胞，已知通过噬菌体、抗原表达和细胞因子释放对组织损伤作出反应。新出现的证据已经牵连到背根神经性巨噬细胞对神经损伤背景下的神经病痛发展和斧道修复的贡献。快速的声叫（或"快速隔离"），背根神经质噬细胞在神经损伤的情况下的反应需要识别未知的神经免疫因素。在这里，我们演示了我们的实验室如何使用无酶的机械解离方案快速有效地从背根结节分离巨噬细胞。样品一直保存在冰上，以限制细胞压力。与标准酶协议相比，该协议的耗时要少得多，并常用于荧光激活细胞分拣分析。

为了验证分离的细胞，我们首先选择了巨噬细胞Fas诱导凋亡（MAFIA）转基因小鼠17。此行表示在CSF1受体（CSF1R）的启动子（CSF1R）的调控下，一种药物诱导的FK506结合蛋白（FKBP）-Fas自杀融合基因和绿色荧光蛋白（eGFP），这种蛋白在巨噬细胞和微胶质中具体表达。系统注射FK结合蛋白二聚体，AP20187（AP），诱导表达转基因的细胞凋亡。EGFP的表达还允许我们监测DRG中的巨噬细胞。为?...

Watch this Scientific Journal Video about Rapid Isolation of Dorsal Root Ganglion Macrophages at JoVE.com

Rapid Isolation of Dorsal Root Ganglion Macrophages

在这里，我们提出了一个机械分离协议，以快速分离巨噬细胞从背根结条，用于方位和功能分析。

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 ...

新出现的证据涉及背根结节巨噬细胞对神经病变疼痛发育和神经损伤背景下的斧尾修复的贡献。快速表型的后根结节巨噬细胞的反应是需要识别未知的神经免疫因素。在这里，我们演示我们的实验室如何使用无酶的机械分离协议快速有效地将巨噬细胞从后根根刚体分离。与标准酶协议相比，该协议的耗时要小很多，而且我们经常把它用于荧光激活细胞分选分析。在开始实验之前，准备密度梯度介质的工作溶液，将九卷介质与一卷无钙和无镁的 10X HBSS 混合，并放在冰上。确认鼠标已完全麻醉后爪捏没有反应。首先将麻醉小鼠放在化学烟罩内，用胶带固定四只爪子。使用钳子将皮肤抬到肋骨下方。然后用手术剪刀，做一个小切口，露出肝脏和隔膜。使用相同的剪刀，切割隔膜和肋骨。然后打开胸腔，露出跳动的心脏。接下来用虹膜剪刀，快速剪右心房附属物。一旦出血被注意到，插入一个30规格的针头到左心室的后端，并慢慢注射10毫升预冷却的1XPBS，以穿行动物。要进行后视乳切除术，请将鼠标放在易发位置。使用大小 11 手术刀，使两个纵向横向深切口从胸腔区域开始到神圣区域。然后去除露出背部肌肉层的皮肤。然后与弗里德曼-皮尔森荣弗，剥离结缔组织和肌肉，直到隆椎过程和双边横向过程暴露。首先，使用弗里德曼-皮尔森的脊柱小心地打开后腰脊柱，然后在弗里德曼-皮尔森荣格和诺耶斯弹簧剪刀之间切换，以去除椎骨，露出脊髓与完整的脊髓神经连接。小心解剖异质和反向腰椎 DRG。放入一毫升冰冷钙和无镁1X HBSS中，放入Dounce组织均质器中。在 Dounce 均质器中，用松散的害虫使 DRG 组织均质 20 到 25 次。将无菌 70 微米尼龙细胞过滤器放在无菌的 50 毫升锥形管中。使用 800 微升的冰码 1X HBSS 来弄湿细胞过滤器。使用移液器将均质组织悬浮液从均质器收集到湿细胞过滤器上，并收集到 50 毫升锥形管中。用800微升的冰1X HBSS冲洗两次均质剂，将液体收集到同一个50毫升锥形管中，以增加产量。将1.5毫升的均衡冰冷同位素密度梯度介质加入无菌的五毫升聚苯乙烯烧瓶管中。将细胞均质从50毫升锥形管转移到这种烧瓶管中。和移液器上下混合好。再添加 500 微升 1X HBSS，以密封顶部。在4摄氏度下，将800倍G的细胞离心20分钟。小心吸气介质中含有麦林的上清液，而不干扰管底的细胞颗粒。将含有5%胎儿牛血清的100微升PBS加入颗粒中，并重新悬浮D组细胞。然后将α小鼠CX3CR1APC抗体加入细胞，在黑暗中在4摄氏度下孵育一小时。用五毫升PBS清洗细胞一次。在摄氏4度下将细胞在360倍G下离心8分钟。吸制上一液，然后将细胞颗粒重新悬浮在300微升PBS中，进行事实分析。在将FK结合蛋白二联剂AP注射到MaFIA小鼠以消耗巨噬细胞后，与受控制的车辆治疗小鼠相比，在AP治疗小鼠的DAG中，GFP阳性细胞明显丧失。事实分析表明，在AP治疗的小鼠中，GFP高种群的耗竭成功，并证明了分离细胞的高质量。车辆处理动物 DRG 中 4% 的分离细胞是 GFP 高巨噬细胞。相比之下，在AP治疗的小鼠中，只有0.4%的 DRG 细胞是 GFP 高巨噬细胞。从野生型小鼠机械分离的 DRG 细胞被染有 alpha 小鼠 CX3CR1-APC 抗体。6%的 DRG 细胞为 CX3CR1 阳性巨噬细胞。当用碘化丙二基评估细胞的生存能力时，它揭示了超过80%的新鲜分离的 DRG 细胞是可行的。如果注意到不满意的细胞产量，可能怀疑DG组织不充分或过度均质。此外，DRG 解剖可能需要初学者反复练习。我们的协议的应用可能可以扩展，以研究其他非神经元细胞，如卫星细胞和T细胞。进一步的研究可能需要确认细胞分离的有效性。

Research

JoVE Journal

Neuroscience

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 ...

科研

神经科学

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 ...

Hydraulic Extrusion of the Spinal Cord and Isolation of Dorsal Root Ganglia in Rodents

背根神经节的啮齿动物的脊髓和隔离的液压挤压

Traditionally, the spinal cord is isolated by laminectomy, i.e. by breaking open the spinal vertebrae one at a time. This is both time consuming and may ...

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 ...

Dorsal Root Ganglia Isolation and Primary Culture to Study Neurotransmitter Release

背根神经节分离与原代培养对神经递质释放的研究

Dorsal root ganglia (DRG) contain cell bodies of sensory neurons. This type of neuron is pseudo-unipolar, with two axons that innervate peripheral ...

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

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

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 ...

Use of In Vivo Single-fiber Recording and Intact Dorsal Root Ganglion with Attached Sciatic Nerve to Examine the Mechanism of Conduction Failure

使用 Vivo 单纤维记录和带附骨神经的固定背道根结条，以检查传导失败机制

Single-fiber recording has been a classical and effective electrophysiological technique over the last few decades because of its specific application for ...