Name: adeno-associated virus-mediated transgene expression in genetically defined neurons of the spinal cord
Uploaded: 2018-05-12T13:00:00
Description: Watch this Scientific Journal Video about Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord at JoVE.com

我们感谢 Hanns Zeilhofer 慷慨地支持这项工作。亨德里克 Wildner 得到了奥尔加 Mayenfisch 基金会的支持。我们感谢卡门 Birchmeier 的 Lmx1b 抗体。

所有动物实验均经瑞士州兽医局 (苏黎世) 批准, 并符合所有相关的规章制度和机构准则。
注: 所有材料连同各自的制造商和/或供应商在材料表中列出。
1. 产生依赖于 AAV 的向量
注意: 可以购买各种与不同启动子的依赖关系的向量 (请参见材料表), 或者, 如果所需的表达式构造不可用, 则可以通过修改现有的 AAV ?...

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(73), e50313 (2013).</li><li>Kohro, 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=A+new+minimally-invasive+method+for+microinjection+into+the+mouse+spinal+dorsal+horn.">A new minimally-invasive method for microinjection into the mouse spinal dorsal horn.</a> Sci Rep. 5, 14306 (2015).</li><li>Bourane, S., 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=Gate+control+of+mechanical+itch+by+a+subpopulation+of+spinal+cord+interneurons.">Gate control of mechanical itch by a subpopulation of spinal cord interneurons.</a> Science. 350 (6260), 550-554 (2015).</li><li>Bourane, S., 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=Identification+of+a+spinal+circuit+for+light+touch+and+fine+motor+control.">Identification of a spinal circuit for light touch and fine motor control.</a> Cell. 160 (3), 503-515 (2015).</li><li>Duan, B., 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=Identification+of+spinal+circuits+transmitting+and+gating+mechanical+pain.">Identification of spinal circuits transmitting and gating mechanical pain.</a> Cell. 159 (6), 1417-1432 (2014).</li><li>Gutierrez-Mecinas, 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=Preprotachykinin+A+is+expressed+by+a+distinct+population+of+excitatory+neurons+in+the+mouse+superficial+spinal+dorsal+horn+including+cells+that+respond+to+noxious+and+pruritic+stimuli.">Preprotachykinin A is expressed by a distinct population of excitatory neurons in the mouse superficial spinal dorsal horn including cells that respond to noxious and pruritic stimuli.</a> Pain. 158 (3), 440-456 (2017).</li><li>Awatramani, R., Soriano, P., Rodriguez, C., Mai, J. 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椎管内注射自动增值服务可能成为一个强大的技术在一个研究实验室, 使分析脊髓细胞具有较高的时间和空间的解决方案。该协议使三主要脊柱段支配由感觉神经元扩展其外围轴突到后肢的传导。传感三段产生健壮和可重复的行为数据。它还能在单椎管内注射后检测出更大的感官区域。例如, 相同的注射机制允许测试爪子 (冯弗雷, 哈格里夫斯,等) 和大腿 (皮下注射, 机械受体刺激), 从而扩大?...

背脊髓是人体和大脑周围信息交换的必要条件。感官刺激, 如热, 冷, 接触, 或有害刺激检测的专门的外围神经元, 这将信息传递给脊髓背角神经元。在这里, 一个复杂的抑制和兴奋中间神经元网络调节并最终通过脊髓投射神经元传递感官信息到 supraspinal 站点 1, 2.由脊柱间和投射神经元之间进行的计算, 以此来确定哪些信息被抑制或中继到何种强度。感官刺激的整合的变化, 如抑制和刺激之间的平衡改变, 可能导致感官功能障碍, 如过敏性或痛觉 (通常不痛刺激后的痛苦感觉)。这些更改被认为是各种慢性疼痛状态的根本原因3,4。因此, 脊柱电路在感官加工中具有高度的重要性, 因此在感知有机体的环境和自我方面是非常重要的。随着分子、遗传和外科技术的最近出现和结合, 使基因识别的脊髓神经元亚群精确操作, 科学家们现在开始了解底层的脊柱电路负责不同感官方式的加工。
椎管内注射 rAAV 为野生型或转基因小鼠, 极大地促进了对脊髓神经元特定子集功能的操作、分析和理解5,6,7,8,9,10,11. 此技术允许在空间上提供标记蛋白 (如 gfp/gfp 融合蛋白)、报告蛋白 (如 GCaMP) 或效应蛋白 (如细菌毒素、channelrhodopsin 或遗传药理学受体)。脊髓神经元受限的方式。局部注射 rAAVs 依赖于转基因小鼠, 表达 recombinase 在脊髓神经元的特定子集, 允许对各自神经元种群的具体分析。我们使用这种技术来标记、消融、抑制或激活脊髓 glycinergic 神经元, 证明它们是控制疼痛和瘙痒传输的脊髓门的重要部分7。在这些实验中, 椎管内注射 rAAV 为 GlyT2::Cre 小鼠, 使脊髓 glycinergic 神经元有选择性的操作。因此, 可以避免同时操作包含对动物生存至关重要的 glycinergic 神经元的 supraspinal 电路。
虽然椎管内注射 rAAVs 限制了对注射部位的感染, 但病毒转导不仅可以发生在局部神经元中, 而且也可能出现在通过轴突投射连接到注射部位的神经元中。后者经常被用来追踪中枢神经系统的区域, 为大脑中特定的细胞核提供神经元输入。然而, 轴突投射的感染也可能是一个混淆的因素, 当一个定义的神经元群体应在特定的地点进行研究。为了解决这些问题, 我们最近对 AAV 血清型和表达盒进行了全面分析, 以确定血清型和促进剂, 可用于最小化或最大限度地逆行转导。本文在脊柱电路研究的背景下, 分析了不同血清型和促进剂对背根神经节、延髓丘脑髓质 (RVM) retrogradely 传感器神经元的能力, 以及体感皮层12. 因此, 本议定书中概述的技术可以用于分析注射部位的脊髓神经元, 也可用于分析为脊髓注入部位提供输入的投射神经元。在这里描述的协议中, 三注射 rAAV 到腰脊髓左侧, 使三腰椎段 (L3-L5) 神经元转导。L3-L5 段接受大部分的感觉输入从后肢同侧到注射部位。我们证明, L3-L5 基因标记神经元的功能操作足以唤起强健的行为变化, 从而为基因标记的神经元亚型的电路功能提供功能性证据。

<table>
	<tbody>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>micropipette puller: DMZ-Universal-Electrode-Puller</td>
			<td>Zeitz</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>anesthesia unit: Oxymat3 oxygen concentrator</td>
			<td>Weinmann</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>anesthesia unit: VIP 3000 Veterinary Vaporizer</td>
			<td>Midmark</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat mat: Mio Star Thermocare 100</td>
			<td>Migros</td>
			<td>717614700000</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric shaver</td>
			<td>Philips</td>
			<td>BT9290</td>
			<td></td>
		</tr>
		<tr>
			<td>surgical microscope (OPMI pico)</td>
			<td>Zeiss</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Small animal stereotaxic apparatus</td>
			<td>Kopf</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Neurostar StereoDrive (optional)</td>
			<td>Neurostar</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Model 51690 Cunningham mouse spinal adaptor</td>
			<td>Harvard Apparatus</td>
			<td>72-4811</td>
			<td></td>
		</tr>
		<tr>
			<td>PHD Ultra syringe pump with nanomite</td>
			<td>Harvard Apparatus</td>
			<td>70-3601</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton 701 RN 10 &#956;l glass microliter syringe</td>
			<td>Hamilton</td>
			<td>7635-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton Removable needle (RN) compression fitting 1 mm</td>
			<td>Hamilton</td>
			<td>55750-01</td>
			<td></td>
		</tr>
		<tr>
			<td>fine dentistry drilling apparatus: Osada success 40</td>
			<td>Osada</td>
			<td>OS-40</td>
			<td></td>
		</tr>
		<tr>
			<td>spherical cutter, 0.5mm</td>
			<td>Busch</td>
			<td>12001005B</td>
			<td></td>
		</tr>
		<tr>
			<td>electronic von Frey anesthesiometer</td>
			<td>IITC</td>
			<td>23905</td>
			<td></td>
		</tr>
		<tr>
			<td>flexible von Frey hairs</td>
			<td>IITC</td>
			<td>#7</td>
			<td></td>
		</tr>
		<tr>
			<td>LSM710 Pascal confocal microscope</td>
			<td>Zeiss</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>0.8 NA &#215; 20 Plan-apochromat objective</td>
			<td>Zeiss</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>1.3 NA &#215; 40 EC Plan-Neofluar oil-immersion objective</td>
			<td>Zeiss</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Surgical Tools</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Handle #4, 13cm</td>
			<td>Fine Science Tools</td>
			<td>10004-13</td>
			<td></td>
		</tr>
		<tr>
			<td>Extra Fine Bonn Scissors</td>
			<td>Fine Science Tools</td>
			<td>14084-08</td>
			<td></td>
		</tr>
		<tr>
			<td>Adson forceps, 1 x 2 teeth, 12 cm</td>
			<td>Fine Science Tools</td>
			<td>11027-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Friedman-Pearson rongeurs, curved, 0.7 mm cup</td>
			<td>Fine Science Tools</td>
			<td>16121-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #2 laminectomy forceps</td>
			<td>Fine Science Tools</td>
			<td>11223-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Olsen-Hegar needle holders, serrated, 8.5 mm clamp length</td>
			<td>Fine Science Tools</td>
			<td>12002-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine forceps #5</td>
			<td>Fine Science Tools</td>
			<td>11254-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Consumables and Chemicals</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Thin-wall glass capillary, 1mm outside diameter</td>
			<td>World Precision Instruments</td>
			<td>TW 100-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringes (1, 5 and 20 ml)</td>
			<td>B. Braun</td>
			<td>(9166917V, 4606051V, 4606205V)</td>
			<td></td>
		</tr>
		<tr>
			<td>26G beveled needle</td>
			<td>B. Braun</td>
			<td>4665457</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile scalpel blades</td>
			<td>B. Braun</td>
			<td>BB523</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical sutures Safil Quick+ 4/0, absorbable</td>
			<td>B. Braun</td>
			<td>C1046220</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical sutures Premilene 5/0, non-absorbable</td>
			<td>B. Braun</td>
			<td>C0932191</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile PBS or saline (0.9%)</td>
			<td></td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol, 70% (disinfectant)</td>
			<td></td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Iodine solution (e.g. Braunol)</td>
			<td>B. Braun</td>
			<td>18380</td>
			<td></td>
		</tr>
		<tr>
			<td>Anaesthetics&#160; (e.g. Attane isoflurane)</td>
			<td>Provet</td>
			<td>2222</td>
			<td></td>
		</tr>
		<tr>
			<td>Aldasorber</td>
			<td>Provet</td>
			<td>333526</td>
			<td></td>
		</tr>
		<tr>
			<td>analgesics (e.g. buprenorphine: temgesic)</td>
			<td>Indivior</td>
			<td>GTIN: 7680419310018</td>
			<td></td>
		</tr>
		<tr>
			<td>Ophthalmic ointment (e.g. vita-pos)</td>
			<td>Pharma medica</td>
			<td>GTIN: 4031626710635</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swabs (e.g. from)</td>
			<td>IVF Hartmann</td>
			<td>1628100</td>
			<td></td>
		</tr>
		<tr>
			<td>Facial tissues (e.g. from)</td>
			<td>Uehlinger AG</td>
			<td>2015.10018</td>
			<td></td>
		</tr>
		<tr>
			<td>Superfrost plus microscope slides</td>
			<td>ThermoScientific</td>
			<td>J1800AMNZ</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Mice</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6J mice&#160; (wildtype)</td>
			<td>The Jackson Laboratory</td>
			<td>RRID:IMSR_JAX:000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Rorbtm1.1(cre)Hze/J mice (ROR&#946;Cre)</td>
			<td>The Jackson Laboratory</td>
			<td>RRID:IMSR_JAX:023526</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J mice (R26Tom)</td>
			<td>The Jackson Laboratory</td>
			<td>RRID: IMSR_JAX:007914</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Viral vectors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>AAV1.CB7.CI.eGFP.WPRE.rBG (AAV1.CAG.eGFP)</td>
			<td>Penn Vector Core</td>
			<td>AV-1-PV1963</td>
			<td></td>
		</tr>
		<tr>
			<td>AAV1.CAG.flex.eGFP.WPRE.bGH (AAV1.CAG.flex.eGFP)</td>
			<td>Penn Vector Core</td>
			<td>AV-1-ALL854</td>
			<td></td>
		</tr>
		<tr>
			<td>AAV1.CAG.flex.tdTomato.WPRE.bGH (AAV1.CAG.flex.tdTomato)</td>
			<td>Penn Vector Core</td>
			<td>AV-1-ALL864</td>
			<td></td>
		</tr>
		<tr>
			<td>AAV1.EF1a.flex.DTA.hGH (AAV1.EF1a.flex.DTA)</td>
			<td>Penn Vector Core</td>
			<td>Custom production</td>
			<td></td>
		</tr>
		<tr>
			<td>AAV1.hSyn.DIO.hM3D(Gq)-mCherry.hGH (AAV.flex.hM3D(Gi))</td>
			<td>Penn Vector Core</td>
			<td>Custom production</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Plasmids</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>pAAV.hSyn.flex.hM3D(Gq)-mCherry</td>
			<td>Addgene</td>
			<td>44361</td>
			<td></td>
		</tr>
		<tr>
			<td>pAAV.EF1&#945;.flex.hChR2(H134R)-eYFP</td>
			<td>Addgene</td>
			<td>20298</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Bacteria</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MDS42</td>
			<td>ScarabGenomics</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stbl3</td>
			<td>ThermoScientific</td>
			<td>C737303</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Reagents</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>EndoFree Plasmid Maxi Kit</td>
			<td>Quiagen</td>
			<td>12362</td>
			<td></td>
		</tr>
		<tr>
			<td>NucleoBond PC 500</td>
			<td>Machery &#38; Nagel</td>
			<td>740574</td>
			<td></td>
		</tr>
		<tr>
			<td>clozapine-N-oxide (CNO)</td>
			<td>Enzo Life Sciences</td>
			<td>BBL-NS105-0025</td>
			<td></td>
		</tr>
		<tr>
			<td>chloroquine diphosphate salt</td>
			<td>Sigma</td>
			<td>C6628</td>
			<td></td>
		</tr>
		<tr>
			<td>histamine</td>
			<td>Sigma</td>
			<td>H7125</td>
			<td></td>
		</tr>
		<tr>
			<td>Dapi</td>
			<td>Invitrogen</td>
			<td>D3571</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Antibodies (dilution)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rabbit anti-GFP (1:1000)</td>
			<td>Molecular Probes</td>
			<td>RRID:AB_221570</td>
			<td></td>
		</tr>
		<tr>
			<td>Rabbit anti-NeuN (1:3000)</td>
			<td>Abcam</td>
			<td>RRID:AB_10711153</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-Pax2 (1 : 200)</td>
			<td>R &#38; D Systems</td>
			<td>RRID:AB_10889828</td>
			<td></td>
		</tr>
		<tr>
			<td>Guinea pig anti-Lmx1b (1 : 10 000)</td>
			<td>Dr Carmen Birchmeier</td>
			<td>Muller et al. 2002</td>
			<td></td>
		</tr>
		<tr>
			<td>Rabbit anti-GFAP (1 : 1000)</td>
			<td>DakoCytomation</td>
			<td>RRID:AB_10013382</td>
			<td></td>
		</tr>
		<tr>
			<td>Secondary antibodies raised in donkey (1:800)</td>
			<td>Jackson ImmunoResearch Laboratories</td>
			<td>NA</td>
			<td></td>
		</tr>
	</tbody>
</table>

adeno-associated virus-mediated transgene expression in genetically defined neurons of the spinal cord

脊髓神经元亚群的选择性操纵主要是通过两种不同的方法实现的: 1) 交叉遗传学, 利用双或三种转基因小鼠来实现有选择性表达的记者或效应者。基因 (例如, 从 Rosa26 轨迹) 在期望脊柱人口。2) 椎管内注射重组腺相关病毒 (rAAV);本文对选择的报告者或效应基因进行了 AAV 矢量编码, 并将其用于表达 recombinase 在所期望的神经元亚群中的小鼠脊髓。该协议描述了如何产生 rAAV 载体和如何传感器脊髓背角 L3-L5 与 rAAVs 神经元。由于腰椎段 L3-L5 是由那些从后肢传递感官信息的外周感觉神经元支配的, 自发的行为和对向侧向一侧与注射侧的感觉测试的反应, 可以分析了操作神经元在感官处理中的作用。我们提供的例子, 这项技术可以用来分析基因定义的脊髓神经元子集。在转基因小鼠中, 病毒介导的转基因基因表达的主要优势与经典的小鼠转基因基因表达相比有以下特点: 1) 不同的 rAAVs 编码各种记者或效应蛋白可注入单一的转基因生产线, 从而克服了创建多个转基因小鼠线的需要。2) 椎管内注射限制了对注射部位和注射后的时间对细胞的操作。主要缺点有: 1) rAAVs 的报告基因表达更具变数。2) 手术是需要传感器脊髓神经元的兴趣。这两种方法中哪一种比较合适取决于神经元的数量和要处理的研究问题。

为了说明椎管内注射 rAAV 编码标记蛋白所能获得的表达水平, 我们首先注射了 AAV1。eGFP 进入野生型小鼠的腰椎脊髓。三个注射间隔约1毫米, 产生了几乎连续感染腰椎段 L3 到 L5 (图 1A-c)。病毒注射在深度300µm 从脊柱表面导致主要感染的细胞在脊髓背角。但是, 也可以在腹角 (图 1D) 中找到受感染的细胞。下一步, 目的?...

Watch this Scientific Journal Video about Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord at JoVE.com

Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord

腺相关病毒介导的脊髓基因神经元的转基因表达

椎管内注射 recombinase 依赖重组腺相关病毒 (rAAV) 可以用来操纵任何基因标记的细胞类型的脊髓。在这里, 我们描述如何传感器神经元的背角的腰椎脊髓。这种技术可以对纵的神经元亚型进行功能性审问。

Selective manipulation of spinal neuronal subpopulations has mainly been achieved by two different methods: 1) Intersectional genetics, whereby double or ...

Research

JoVE Journal

Neuroscience

Dissection and Culture of Commissural Neurons from Embryonic Spinal Cord

从胚胎脊髓​​连合神经的解剖和文化

Commissural neurons have been widely used to investigate the mechanisms underlying axon guidance during embryonic spinal cord development. The cell bodies ...

科研

神经科学

Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication

脊髓电二：外吸电极的制作

Development of neural circuitries and locomotion can be studied using neonatal rodent spinal cord central pattern generator (CPG) behavior. We demonstrate ...

Viral Tracing of Genetically Defined Neural Circuitry

病毒遗传背景明确的神经回路跟踪

Classical methods for studying neuronal circuits are fairly low throughput. Transsynaptic viruses, particularly the pseudorabies (PRV) and rabies virus ...

Spinal Cord Transection in the Larval Zebrafish

在斑马鱼幼虫脊髓横断

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability ...

RNA-Seq Analysis of Differential Gene Expression in Electroporated Chick Embryonic Spinal Cord

对基因差异表达的RNA序列中电穿孔鸡胚脊髓分析

In ovo electroporation of the chick neural tube is a fast and inexpensive method for identification of gene function during neural development. Genome ...

Diffusion Imaging in the Rat Cervical Spinal Cord

在大鼠颈脊髓弥散成像

Magnetic resonance imaging (MRI) is the state of the art approach for assessing the status of the spinal cord noninvasively, and can be used as a ...

Zebrafish In Situ Spinal Cord Preparation for Electrophysiological Recordings from Spinal Sensory and Motor Neurons

Zebrafish In Situ Spinal Cord Preparation for Electrophysiological Recordings from Spinal Sensory and Motor Neurons

斑马鱼<em&gt;原位</em&gt;脊髓制备用于脊髓感觉和运动神经元的电生理记录

Zebrafish, first introduced as a developmental model, have gained popularity in many other fields. The ease of rearing large numbers of rapidly developing ...

Spinal Cord Neurons Isolation and Culture from Neonatal Mice

脊髓神经元从新生小鼠中分离和培养

We present a protocol for the isolation and culture of spinal cord neurons. The neurons are obtained from neonatal C57BL/6 mice and are isolated on ...

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

胸腺1-gmcmp6 转基因小鼠在原代体血源皮质中的成像神经活性

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in ...

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

神经干细胞向大鼠和小鼠大脑的动脉内传递：对脑缺血的应用

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...