Name: in vivo electrophysiological measurement of the rat ulnar nerve with axonal excitability testing
Uploaded: 2018-02-06T12:30:00
Description: Watch this Scientific Journal Video about In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing at JoVE.com

该项目得到了 Lundbeck 基金会、诺德基金会、丹麦医学研究理事会、Ludvig 和萨拉 Elsass 基金会、神经病学研究基金会和 Jytte Kaj 基金会的支持。R. A 得到澳大利亚国家卫生和医学研究理事会 (#1091006) 早期职业博士后奖学金的支持

这里描述的所有实验程序都符合悉尼新南动物保育和伦理委员会的要求, 并按照澳大利亚国家卫生和医学研究理事会 (澳洲) 的规定进行动物实验。
1. 实验设置
注:12 周前, 雌性长埃文斯大鼠被用于此手术。
<ol><li>麻醉在感应室使用4% 异氟醚和1升每分钟2流量。确认充分的麻醉, 通过测试矫正反射, 并确保其缺席之前, 从诱导室移除动物。请注意, 各?...

<ol>
	<li>Krishnan, A. V., Lin, C. S. -. Y., Park, S. B., Kiernan, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axonal+ion+channels+from+bench+to+bedside:+a+translational+neuroscience+perspective.">Axonal ion channels from bench to bedside: a translational neuroscience perspective.</a> Prog neurobiol. 89 (3), 288-313 (2009).</li><li>Lin, C. S. -. Y., Krishnan, A. V., Park, S. B., Kiernan, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulatory+effects+on+axonal+function+after+intravenous+immunoglobulin+therapy+in+chronic+inflammatory+demyelinating+polyneuropathy.">Modulatory effects on axonal function after intravenous immunoglobulin therapy in chronic inflammatory demyelinating polyneuropathy.</a> Arch neurol. 68 (7), 862-869 (2011).</li><li>Park, S. B., Goldstein, D., Lin, C. S. -. 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Transplant. 13 (9), 2426-2432 (2013).</li><li>Bo&#235;rio, D., Greensmith, L., Bostock, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Excitability+properties+of+motor+axons+in+the+maturing+mouse.">Excitability properties of motor axons in the maturing mouse.</a> J. Peripher. Nerv. Syst. 14 (1), 45-53 (2009).</li><li>Bo&#235;rio, D., Kalmar, B., Greensmith, L., Bostock, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Excitability+properties+of+mouse+motor+axons+in+the+mutant+SOD1(G93A)+model+of+amyotrophic+lateral+sclerosis.">Excitability properties of mouse motor axons in the mutant SOD1(G93A) model of amyotrophic lateral sclerosis.</a> Muscle &amp; Nerve. 41 (6), 774-784 (2010).</li><li>Alvarez, S., Calin, A., Graffmo, K. S., Moldovan, M., Krarup, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+motor+axons+of+SOD1(G127X)+mutant+mice+are+susceptible+to+activity-dependent+degeneration.">Peripheral motor axons of SOD1(G127X) mutant mice are susceptible to activity-dependent degeneration.</a> Neurosci. 241, 239-249 (2013).</li><li>Fledrich, 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=Soluble+neuregulin-1+modulates+disease+pathogenesis+in+rodent+models+of+Charcot-Marie-Tooth+disease+1A.">Soluble neuregulin-1 modulates disease pathogenesis in rodent models of Charcot-Marie-Tooth disease 1A.</a> Nat. 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V., Morris, R., Krarup, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nerve+excitability+in+the+rat+forelimb:+a+technique+to+improve+translational+utility.">Nerve excitability in the rat forelimb: a technique to improve translational utility.</a> J. Neurosci. Methods. 275, 19-24 (2017).</li><li>Moldovan, M., Alvarez, S., Krarup, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+axon+excitability+during+Wallerian+degeneration.">Motor axon excitability during Wallerian degeneration.</a> Brain. 132 (Pt 2), 511-523 (2009).</li><li>Madison, R. D., Robinson, G. A., Krarup, C., Moldovan, M., Li, Q., Wilson, W. 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所描述的过程证明了一种简单而可靠的微创技术, 允许在短时间内对轴突的生物物理特性和膜电位进行评估。与其他更多的侵入性技术, 需要暴露的神经, 目前的方法轴突兴奋性测试诱导最小的组织损害, 从而使在体内评估, 保留了生理条件神经的兴趣, 并允许重复测量。
为了确保取得一致的结果, 有一些方法上的考虑需要加以解决。其中一个因素是麻醉的浓度和类型。以...

使用电生理学技术是一个必要的工具, 在体内研究周围神经功能的神经紊乱。传统的神经传导方法利用 supramaximal 刺激记录运动动作电位振幅和潜伏期。因此, 这些技术提供了关于导电纤维数量和最快纤维的传导速度的有用信息。一个有价值的辅助工具是轴突兴奋性测试。这种技术使用复杂的电生理刺激模式间接评估周围神经的生物物理特性, 如离子通道、能量依赖性泵、离子交换过程和膜电位的活性1。
轴突兴奋性测试通常用于临床设置, 以调查病理生理过程和治疗干预对各种神经疾病的影响。重要的是, 轴突兴奋性措施是敏感的治疗干预, 影响周围神经功能, 如静脉注射免疫球蛋白 (丙种球蛋白) 治疗2, 化疗3和磷酸抑制剂 (CNI) 治疗4. 虽然这些研究提供了重要的见解, 但临床研究往往排除了早期疾病特征的调查和新的治疗方案5。因此, 这些方法在神经紊乱动物模型中的应用最近获得了牵引6,7,8,9。事实上, 这些方法提供了一个机会, 以了解特定的神经兴奋性变化与这些疾病相关, 从而推进转化研究。
这里描述的程序是一种简单而可靠的方法来记录轴突兴奋性措施的完整大鼠尺神经。

<table>
	<tbody>
		<tr>
			<td>QTracS Program</td>
			<td>Digitimer Ltd.</td>
			<td></td>
			<td>Axonal excitability program</td>
		</tr>
		<tr>
			<td>AM-Systems 2200, Analog Stimulus Isolator, 2200V/50Hz</td>
			<td>SDR Scientific</td>
			<td>850005</td>
			<td>Stimulator</td>
		</tr>
		<tr>
			<td>High Performance AC Amplifier Model LP511</td>
			<td>Grass Technologies</td>
			<td></td>
			<td>Amplifier</td>
		</tr>
		<tr>
			<td>Humbug 50/60Hz Noise eliminator</td>
			<td>Quest Scientific Instruments</td>
			<td>726310</td>
			<td>Noise eliminator</td>
		</tr>
		<tr>
			<td>Low Impedance Platinum Monopolar Subdermal Needle Electrodes</td>
			<td>Grass Technologies</td>
			<td>F-E2-24</td>
			<td>Recording electrodes, 10 mm length, 30 gauge</td>
		</tr>
		<tr>
			<td>Low Impedance Platinum Electroencephalography Needle Electrodes</td>
			<td>Cephalon</td>
			<td>9013L0702</td>
			<td>Stimulating electrodes, 10 mm length, 30 gauge</td>
		</tr>
		<tr>
			<td>Multifunction I/O Device Model USB-6341</td>
			<td>National Instruments</td>
			<td></td>
			<td>Multifunction input/output device</td>
		</tr>
		<tr>
			<td>Iron Base Plate IP</td>
			<td>Narishige Scientific Instrument Laboratory</td>
			<td></td>
			<td>Used for holding stimulating needle electrode in place</td>
		</tr>
		<tr>
			<td>Rotating X-block X-4</td>
			<td>Narishige Scientific Instrument Laboratory</td>
			<td></td>
			<td>Used for holding stimulating needle electrode in place</td>
		</tr>
		<tr>
			<td>Magnetic Stand GJ-8</td>
			<td>Narishige Scientific Instrument Laboratory</td>
			<td></td>
			<td>Used for holding stimulating needle electrode in place</td>
		</tr>
		<tr>
			<td>Micromanipulator M-3333</td>
			<td>Narishige Scientific Instrument Laboratory</td>
			<td></td>
			<td>Used for holding stimulating needle electrode in place</td>
		</tr>
	</tbody>
</table>

in vivo electrophysiological measurement of the rat ulnar nerve with axonal excitability testing

电生理学能够客观评估周围神经功能在体内。传统的神经传导措施, 如振幅和潜伏期检测慢性轴突损失和鞘, 分别。轴突兴奋技术 "通过阈值跟踪" 扩大了这些措施, 提供有关的离子通道, 泵和交换器的活动有关的急性功能和可能之前的退化事件的信息。因此, 在神经紊乱动物模型中使用轴突兴奋性可以为评估新的治疗干预提供一个有用的体内措施。在这里, 我们描述了一个实验设置的多项措施的运动轴突兴奋技术在大鼠尺神经。
动物被麻醉与异氟醚和仔细监测, 以确保持续和充分的麻醉深度。持续监测血液中的体温、呼吸速率、心率和血氧饱和度。轴突兴奋性研究是利用经皮刺激尺神经和记录从鱼际肌肉的前肢爪。在正确的电极放置, 一个明确的复合肌肉动作电位增加振幅随着刺激强度增加记录。然后利用一个自动程序提供一系列的电脉冲, 产生5特定的兴奋性措施在以下序列: 刺激反应行为, 强度持续时间常数, 阈 electrotonus, 电流阈值关系和恢复周期。
这里提供的数据表明, 这些措施是可重复的, 并显示了在同一天评估左、右尺神经的相似性。这些技术的局限性是在麻醉条件下剂量和时间的影响。对这些变数进行仔细的监测和记录应在分析时加以考虑。

采用本方案获得了大鼠尺神经的电生理测量。图 3演示了一个12周大的雌性长埃文斯鼠左尺神经的有代表性的记录。复合肌肉动作电位与同时激活的导电纤维数量有关。supramaximal 峰值响应 (mV) (图 3A) 演示了在增量递增刺激之前达到的峰值响应 (图 2B)。
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Watch this Scientific Journal Video about In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing at JoVE.com

In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing

在体内轴突兴奋性试验对大鼠尺神经的电生理测量

轴突兴奋技术提供了一个强有力的工具, 以检查病理生理学和生物物理变化之前, 不可逆转的退化事件。这篇手稿展示了这些技术在麻醉大鼠尺神经上的应用。

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In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing

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