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

プロジェクトは、ルンドベック財団、ノボ ノルディスク財団、デンマークの医学研究評議会、Ludvig とサラ Elsass 財団、イテ ・神経研究財団とカイ Dahlboms 財団によって支えられました。R.A 国立保健医療研究評議会のオーストラリア (#1091006) の初期のキャリア ドクトラル交わりに支えられて

ここで説明したすべての実験手順は動物愛護及びシドニー ニューサウス ウェールズ大学の倫理委員会に適合し、国民の健康と動物実験のためのオーストラリアの規則の医学研究評議会 (NHMRC) に従い行った。
1. 実験のセットアップ
注: この手順では 12 週齢、女性のロング ・ ラットが使用されました。
<ol><li>分 O2流量あたり 4% イソフルラン...

<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. -. Y., Krishnan, A. V., Friedlander, M. L., 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=Acute+abnormalities+of+sensory+nerve+function+associated+with+oxaliplatin-induced+neurotoxicity.">Acute abnormalities of sensory nerve function associated with oxaliplatin-induced neurotoxicity.</a> J. Clin. Oncol. 27 (8), 1243-1249 (2009).</li><li>Arnold, R., Pussell, B. A., Pianta, T. J., Lin, C. S. -. Y., Kiernan, M. C., Krishnan, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+between+calcineurin+inhibitor+treatment+and+peripheral+nerve+dysfunction+in+renal+transplant+recipients.">Association between calcineurin inhibitor treatment and peripheral nerve dysfunction in renal transplant recipients.</a> Am. J. 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. Med. 20 (9), 1055-1061 (2014).</li><li>Vianello, 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=Low+doses+of+arginine+butyrate+derivatives+improve+dystrophic+phenotype+and+restore+membrane+integrity+in+DMD+models.">Low doses of arginine butyrate derivatives improve dystrophic phenotype and restore membrane integrity in DMD models.</a> FASEB J. 28 (6), 2603-2619 (2014).</li><li>Osaki, 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=Effects+of+anesthetic+agents+on+in+vivo+axonal+HCN+current+in+normal+mice.">Effects of anesthetic agents on in vivo axonal HCN current in normal mice.</a> Clin Neurophysiol. 126 (10), 2033-2039 (2015).</li><li>Biessels, G. 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=Phenotyping+animal+models+of+diabetic+neuropathy:+a+consensus+statement+of+the+diabetic+neuropathy+study+group+of+the+EASD+(Neurodiab).">Phenotyping animal models of diabetic neuropathy: a consensus statement of the diabetic neuropathy study group of the EASD (Neurodiab).</a> J. Peripher. Nerv. Syst. 19 (2), 77-87 (2014).</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=A+model+of+mouse+motor+nerve+excitability+and+the+effects+of+polarizing+currents.">A model of mouse motor nerve excitability and the effects of polarizing currents.</a> J. Peripher. Nerv. Syst. 16 (4), 322-333 (2011).</li><li>Arnold, R., Moldovan, M., Rosberg, M. R., Krishnan, A. 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. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+electrophoresis+and+in+vivo+electrophysiology+of+peripheral+nerve+using+DC+field+stimulation.">In vitro electrophoresis and in vivo electrophysiology of peripheral nerve using DC field stimulation.</a> J. Neurosci. Methods. 225, 90-96 (2014).</li><li>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=Evaluation+of+Na+/K++pump+function+following+repetitive+activity+in+mouse+peripheral+nerve.">Evaluation of Na+/K+ pump function following repetitive activity in mouse peripheral nerve.</a> J. Neurosci. Methods. 155 (2), 161-171 (2006).</li></ol>

記載された手順を示します、シンプルで信頼性の高い、低侵襲技術ができる生物物理物性評価および軸索の膜電位時間の短い期間でです。神経の露出を必要とする他のより侵襲的な技術と比較して軸索興奮性のテスト法誘導の生理状態を維持する体内の評価ができ最小限の組織損傷、興味の神経、繰り返し測定が可能します。
確実に一貫性のある結果が、対処す?...

電気生理学的技術の使用は神経疾患における末梢神経機能の生体内で調査のために不可欠なツールです。従来神経伝導メソッドは、潜時や振幅運動の活動電位を記録する激刺激を利用します。これらの技術は、繊維の実施数と最速の線維の伝導速度したがって有用な情報を提供します。貴重な相補的なツールは、軸索興奮性のテストのことです。この手法は、イオン チャネル、エネルギー依存性ポンプ、イオン交換プロセス、膜電位の活性などの末梢神経の生物物理学的プロパティを直接評価する洗練された電気生理学的刺激のパターンを使用してください。1。
病態生理学的プロセスと様々 な脳神経疾患の治療介入の効果を検討する臨床場面における軸索興奮性テストを利用して一般的。重要なは、軸索興奮性対策2静脈内の免疫グロブリン (IVIg) 療法など末梢神経機能に影響を与える治療に敏感である化学療法3カルシニューリン阻害剤 (CNI) 治療4. 臨床研究がこれらの研究は、重要な洞察を提供している、初期の病の特徴と治療の新しい選択肢5の調査を排除する頻繁。したがって、神経疾患の動物モデルでこれらのメソッドの使用は最近トラクション6,7,8,9を得ています。確かに、これらのメソッドは、トランスレーショナル ・ リサーチを推進したがって、これらの疾患に関連付けられている特定の神経の興奮性の変化を理解する機会を提供します。
ここで説明する手順は、そのままのラットの尺骨神経にレコードの軸索興奮性対策にシンプルで信頼性の高い方法です。

<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

電気生理学末梢神経機能のin vivoの客観的な評価ができます。潜時や振幅検出慢性軸索損失と脱髄、それぞれなど伝統的な神経伝導を測定します。イオン チャンネル、ポンプ、急性機能に関連し、退化的なイベントの前にことがあります交換のアクティビティに関する情報を提供することによってこれらの措置により軸索興奮性技術"しきい値を追跡"を展開します。そのため、神経疾患の動物モデルにおける軸索興奮性の使用新しい治療的介入を評価するために有用な生体内測定があります。ここでラットの尺骨神経で運動軸索興奮性技術の複数の措置の実験装置について述べる。
動物はイソフルラン麻酔、麻酔の定数と十分な深さを確保するため慎重に監視します。体温、呼吸、心拍数、血液中の酸素の飽和を継続的に監視します。軸索興奮性の研究は、尺骨神経と前肢足の研究筋から記録の経皮的刺激を使用して実行されます。正しい電極配置、刺激強度の増加とともに振幅の増加明確な複合筋活動電位が記録されます。自動化されたプログラムを利用する一連の次の順序で 5 特定の興奮性対策を生成する電気パルスを提供する: 刺激応答性状、強度期間時定数、しきい値 electrotonus、電流しきい値関係と回復のサイクル。
ここで表示されるデータは、これらの措置を繰り返し同じ日に評価すると左と右尺骨神経との間の類似性を示すを示します。この設定にこれらの手法の制限は、投与量と麻酔時間の効果です。慎重に監視し、これらの変数の記録は、解析時に考慮のため行わなければなりません。

ラットの尺骨神経の電気生理学的措置は、この議定書が得られました。図 3は、12 週齢雌長いエバンス ラットの左尺骨神経から記録担当者を示します。複合筋活動電位は、同時にアクティブになっている繊維を導電性の数に関連します。激ピーク応答 (mV) (図 3 a) は、(図 2 b) 応答の変化がないまで刺?...

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

生体内で軸索興奮性のテストでラットの尺骨神経の電気生理学的測定

軸索興奮性のテクニックは、病態と不可逆的な変性イベントの前に生体の変化を調べるための強力なツールを提供します。この原稿は、麻酔下ラットの尺骨神経のこれらのテクニックの使用方法を示します。

Electrophysiology enables the objective assessment of peripheral nerve function in vivo. Traditional nerve conduction measures such as amplitude and ...

The overall goal of this technique is to determine a range of excitability properties for the ulnar nerve in living rats. This method can help answer key questions in the field of neuroscience such as, can we measure the pathological changes of nerve function prior to irreversible neurodegenerative events? The main advantage of this technique is that it allows the indirect assessment of peripheral nerve function in vivo enabling repeat measurements for the characterization of disease progression.Before beginning the procedure, confirm the appropriate level of sedation by pedal withdrawal and corneal reflex in a 12-week-old female Long-Evans rat. Place the rat on a feedback controlled heat mat and insert a rectal thermometer probe to maintain the body temperature at 37 degrees Celsius. To record the compound muscle action potential, insert the reference electrode through the dorsal aspect of the fourth digit and the recording electrode through the hypothenar muscle.Place the ground electrode through the skin on the superior aspect of the forearm between the stimulating and recording electrodes. Then insert the percutaneous stimulating needle cathode approximately four millimeters distal to the cubital tunnel at the elbow and insert the anode approximately one centimeter proximally through the skin of the axilla region. Next, use a semi-automated computer controlled axonal excitability program linked to a constant current stimulator and an amplifier to apply a one millisecond square wave pulse to the ulnar nerve with the cathode needle electrode and to record the compound muscle action potential from the hypothenar muscle.Carefully adjust the angle and/or position of the cathode until an optimal biphasic response curve with a consistent amplitude is achieved and stabilize the cathode with a repositionable electrode holder. For threshold tracking, it is critically important that there is a proportionate response to the stimulus. A successful response is indicated by a sigmoid shape in the stimulus response curve.To record a stimulus response curve, incrementally increase the stimulus intensity of a one millisecond impulse until a maximum response is obtained. Record multiple axonal excitability parameters including the threshold electrotonus, current threshold relationship, and recovery cycle. Then transfer the rat into an individual cage with monitoring until full recumbency.The inward and outward rectifying currents can be assessed by depolarizing and hyper polarizing currents in the current voltage parameter. Data regarding the internodal conductances and membrane potential can be obtained by long subthreshold depolarizing and hyper polarizing currents represented by the threshold electrotonus waveform. In this representative experiment, sequential axonal excitability testing on the left and right forelimbs within 35 minutes after the loss of the pedal withdrawal reflex revealed no significant differences between the left and right ulnar nerves in the standard nerve conduction parameters, compound muscle action potential amplitude and latency nor are differences observed in the nerve excitability variables including super excitability and threshold electrotonus hyper polarizing at 90 to 100 milliseconds.Once mastered, this technique can be completed in 20 to 30 minutes if it is performed properly. While attempting this procedure, it is important to take care to place the electrodes in their appropriate positions to ensure an accurate stimulus response curve and successful threshold tracking. After its development, this technique paved the way for researchers in the field of neuroscience to explore the biophysical properties of peripheral nerves in both the clinical setting and in animal models.After watching this video, you should have a good understanding of how to perform a nerve excitability protocol and to obtain a range of biophysical indices in the ulnar nerve.

Research

JoVE Journal

Neuroscience

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

Nerve Excitability Assessment in Chemotherapy-induced Neurotoxicity

化学療法誘発性神経の神経興奮性の評価

Chemotherapy-induced neurotoxicity is a serious consequence of cancer treatment, which occurs with some of the most commonly used chemotherapies1,2. ...

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The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

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モデルとしての嗅覚系は、軸索の成長パターンと形態を研究する<em&gt;インビボ</em

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion of ...

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同時<em&gt; ex vivoで</emによる二つの網膜の&gt;機能テスト<em&gt; in vivoで</em&gt;電図システム

An In vivo electroretinogram (ERG) signal is composed of several overlapping components originating from different retinal cell types, as well as noise ...

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

実験視神経の脱髄のラットモデルにおける視覚誘発電位の記録

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

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の記録細胞内の電圧応答のための電気生理学的手法<em&gt;ショウジョウバエ</em&gt;光刺激に感光体と介在ニューロン<em&gt;インビボ</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

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エレクトロポレーションによる齧歯類坐骨神経内のシュワン細胞へのインビボ遺伝子導入で

The formation of the myelin sheath by Schwann cells (SCs) is essential for rapid conduction of nerve impulses along axons in the peripheral nervous ...

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生体内で1 分子追跡ショウジョウバエ運動神経がシナプス前ターミナルで

An increasing number of super-resolution microscopy techniques are helping to uncover the mechanisms that govern the nanoscale cellular world. ...

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

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生体内で運動ニューロンの変性症のマウスモデルの前肢からの複合筋活動電位の電気生理学的測定

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings ...

Screening of Axonal Degeneration in Carpal Tunnel Syndrome Using Ultrasonography and Nerve Conduction Studies

超音波検査と神経伝導検査を使用して手根管症候群における軸索変性のスクリーニング

Axonal degeneration, indicative of surgical decompression, may coexist in carpal tunnel syndrome (CTS) as the disease progresses. However, the current ...