XHTWは、医学研究でWMケック財団特別若い学者であり、そしてまた、NIH / NHLBIの助成金R01 - HL089598とR01HL091947、および筋ジストロフィー協会の助成金＃69238でサポートされています。この作品は、ハートのCaMKIIシグナリングのための財団Leducqアライアンスによって部分的にサポートされています。 NLは、心臓リズム学会と2009年から2012年米国心臓協会ポスドクフェローシップから心臓ペーシングと電気生理学における2009年から2010年ミシェルMirowski国際フェローシップの受賞者です。

パート1。外科的準備<ol><li>プログラム電気刺激の実験はマウスで生存の手術として行われる場合には無菌状態が必要になります。定期的な明確な手術手技で十分となるしかし、実験の最も一般的なタイプは、自然界にある端末です。 </li><li>マウスは0.5 L /分100％のイソフルラン2％を使用してO 2麻酔です。 </li><li>バリカンは、ネックラインからミッド胸の高さに毛を剃るために使用されます。 </li><li>その手足は、加熱板（インダスインスツルメンツ、ヒューストン、テキサス州）に組み込まれているECG電極の上にテープで麻酔マウスは、仰臥位に配置されます。と外科領域は10％ポビドンヨードで消毒されています。推奨されるシステムは、体温が37.0℃で± 1.0℃に維持されるように、thermoanalyzerシステムによって制御される加熱パッドに接続されて直腸温度プローブが含まれています</li></ol>パート2。右心房と心室にカニュレーションし、EPカテーテルの挿入<ol><li>マウスが完全に麻酔であることをつま先ピンチで確認後、1 / 2インチの切開は、鎖骨のレベルで尾側末端と正中線の右側に行われます。 </li><li>皮下組織、唾液腺、およびリンパ組織は、右頸静脈を可視化する鈍的切開によって分離されています。 </li><li>静脈の近位端は、6から0縫合糸で結ばれている。カテーテルを挿入しながら、この縫合糸を軽く引っ張ると、内頸静脈をまっすぐになります。別の縫合糸は、可視化、セグメントの先端に静脈の下に配置されます。カテーテルが心臓内に最適に配置されていると、この縫合糸は、止血を確実にし、目的の位置（ 図1）にカテーテルを維持するために、カテーテルの周囲に固定されます。 </li><li>コンピュータベースのデータ収集は、現在、表面心電図と同時に心臓内electrograms（すなわち、IOX - 2集録ソフトウェア、Emka技術、VA、米国）の4リード線を記録するために開始されます。心内電位図記録については、カテーテルの電極は、&quot;録音&quot;モード（すなわち、モデルSTG3008、マルチチャンネルシステム、ロイトリンゲン、ドイツ）の外部刺激に接続されています。 </li><li>マイクロはさみを使用して、小さな切開は、静脈の長手方向に作られ、そして1.1F octapolarカテーテル（EPR - 800、ミラーインスツルメンツ、ヒューストン、テキサス州）は右心房に静脈を介して進められる。そっと近位縫合糸を引っ張るとまっすぐに内頸静脈を保つのを助けると右心房と心室にカテーテルを簡単に通過できるようになります。適切なカテーテルの位置は、それぞれ右心室、右心室、房室結節、および右心房のベース、（ 図2）の頂点のレベルで4心臓内electrogramsの波形の可視化によって検証されます。 </li><li>遠位縫合糸は、今カテーテルの位置を確保し、可能な限り出血を防ぐためにオフ結びついています。 </li></ol>第3部。プログラム電気刺激<ol><li>他の電極対は、&quot;録音&quot;モードのままにしながら選択的な心房ペーシングの場合には、アトリウム内にある電極対は、&quot;再コード化&quot;モードから&quot;刺激&quot;モードに切り替えられます。 </li><li>マウスの心房性不整脈に増加脆弱性を持っているかどうかを判断するには、右心房の心臓電気刺激法が実行されます。最初に、心房ペーシングの閾値は、刺激のキャプチャの一貫性をテストするために別の基本的なサイクルの長さ（BCL）で2ミリ秒の電流パルスを（少なくとも50）を適用することによって決定されます。最初のパルス列のBCLは、本質的なBCLよりわずかに低く、そして10ミリ秒（たとえば、100ミリ、90ミリ、80ミリ秒、および70ミリ秒）減少しています。刺激のキャプチャに必要な典型的な電流の振幅は100〜200μAです。 </li><li>洞結節回復時間（SNRT）は100ミリ秒のBCLで15秒心房ペーシングの列車を適用した後に測定されます。 SNRTは、ペーシング電車の中で最後の刺激と最初の自発的な洞性拍動の発症との間の間隔として定義されます。 </li><li>心房有効不応期（AERP）が結合した短いS2早期刺激による固定BCLs（すなわち、100ミリ秒）での心房ペーシングの列車のシリーズを適用することによって決定されます。 S1 - S2間隔は徐々に70msから20msに各ペーシング電車の中で2ミリ秒で減少します。 AERPはS2（普通列車のパルスれS1、およびS2早期刺激）と伝播ビートを生成するために失敗した心房の最長S1 - S2カップリングの間隔として定義されています。各刺激のプロトコルとの間で、少なくとも30秒間の回復期間があった。 </li><li>房室の有効不応期（AV）ノード（AVNERP）が結合したS2早期刺激と100ミリのBCLで心房ペーシングの列車のシリーズを適用することによって決定されます。 S1 - S2間隔は徐々に減少するミリ秒70〜20ミリ〜2ミリ秒ごとにペーシング電車。 AVNERPは、アトリウムに配信早期刺激が彼の潜在的ではなく、QRS波に先行しているで最長のS1 - S2カップリングの間隔として定義されています。各刺激のプロトコルとの間で、少なくとも30秒間の回復期間があった。 </li><li>心房細動（AF）、Verheule らにより記載されたバーストペーシングプロトコルを使用してテストすることを含む心房性不整脈の誘発2 2 2番目のバーストのシリーズは、心房性不整脈の誘発性を決定するために適用されます。最初の2秒のバーストは、40ミリ秒の周期の長さ（CL）を持ち、それぞれの連続する2秒のバーストは、20ミリ秒の最後のCLになるまで、前のバーストより短い2ミリ秒のCLを持っています。不整脈基質の非存在下で、心臓はペーシングプロトコルの直後に洞調律を再開します。心室応答の周波数は、心室の心電図に見られるように、一般に低速であるのに対し、心房粗動の場合には、、心房電位に見られる速い波の規則的なパターンが存在します。心房細動の場合には、表面心電図はP波のない状態で不規則なRR間隔が表示されます。心室の心電図は不規則と遅い心室波（ 図3）を明らかにする一方、さらに、心房電位は、迅速かつ不規則な波が明らかになります。不整脈誘発のプロトコルは、通常三連で行われ、それが3試験の少なくとも2つに誘発することができれば不整脈が存 ​​在すると考えられている。3,4。 </li></ol>パート4。カテーテルの除去<ol><li>すべてのペーシングプロトコルが終了した後、データ収集が停止されます。カテーテルの遠位端の縫合はやさしくカテーテルを解放するために、遮断されます。 </li><li>ターミナルEP研究の場合には、結び目を穏やかにカテーテルを解放するために緩めている。 </li><li>イソフルランの下に頚椎脱臼を使用しながら研究の結論で、マウスを人道的に安楽死されます。 </li></ol>代表的な結果表面心電図と心内electrograms は 、in vivo電気生理学的研究では全体に同時に記録され、そしてすべてのプロトコルの終了後に詳細に検討されています。ベースラインの電気生理学的パラメータには、PR間隔、PQ間隔、QRS幅、QT間隔、およびQRSの形態が含まれています。これらのパラメータは、手動で測定し、または自動的にデータ集録ソフトウェアIOX - 2またはECG - AUTOを（Emka技術、VA、米国）を使用することができます。 SNRT、AERP、およびAVNERPは、洞結節&quot;ペースメーカー&quot;機能、心房、およびAV結節伝導の特性、それぞれに関する情報を提供します。ペーシング誘発性心房細動のエピソードが車路士らによる論文に記載されています。4の例<img src="/files/ftp_upload/1730/1730fig1.jpg" alt="図1" /> 図1：マウスの電気生理学的試験のための心臓内カテーテルのイラスト。A.右内頸静脈を単離し、カニューレを挿入しています。適切なカテーテルの位置が達成されれば静脈の遠位端はオフに関連付けられています。Bは 。マウスは、この手順のため仰臥位に配置されます。C.の漫画は、カテーテルの心臓内の位置を描いた。一対の電極はそれぞれ、右心室、右心室の基部、房室結節、および右心房の頂点のレベルに位置しています。Dは 。 1.1F octapolarカテーテルのアップを閉じます。図は、マートゥルらから変更。サークArrhtyhm Electrophys 5の許可を得て。 <img src="/files/ftp_upload/1730/1730fig2.jpg" alt="図2" /> 図2。マウスの代表的な表面心電図と心内Electrograms。リードIIの毎分540ビートの頻度で定期的な洞調律を示した。（BE）バイポーラ心腔内心電図の記録が右心房（B）のレベルでは、房室結節を設定（C）、とのベースの（A）表面心電図右心室（D）、および右心室の心尖部（E）、それぞれ。パネルBにおける心内膜の波は、表面ECG上でP波に相当することに注意してください。心室の心電図上のV波は表面心電図上のQRS波に相当する。 <img src="/files/ftp_upload/1730/1730fig3.jpg" alt="図3" /> 図3。心房バーストペーシング後心房細動を開発したマウスの代表的な表面心電図と心内electrograms。

<ol>
	<li>Nattel, S., Shiroshita-Takeshita, A., Brundel, B. J., Rivard, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+atrial+fibrillation:+lessons+from+animal+models.">Mechanisms of atrial fibrillation: lessons from animal models.</a> Prog Cardiovasc Dis. 48, 9-9 (2005).</li><li>Verheule, S., Sato, T., Everett, T. t., Engle, S. K., Otten, D., Rubart-von der Lohe, M., Nakajima, H. O., Nakajima, H., Field, L. J., Olgin, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+vulnerability+to+atrial+fibrillation+in+transgenic+mice+with+selective+atrial+fibrosis+caused+by+overexpression+of+TGF-beta1.">Increased vulnerability to atrial fibrillation in transgenic mice with selective atrial fibrosis caused by overexpression of TGF-beta1.</a> Circ Res. 94, 1458-1458 (2004).</li><li>Sood, S., Chelu, M. G., Oort, R. J. v. a. n., Skapura, D., Santonastasi, M., Dobrev, D., Wehrens, X. H., H, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+calcium+leak+due+to+FKBP12.6+deficiency+in+mice+facilitates+the+inducibility+of+atrial+fibrillation.">Intracellular calcium leak due to FKBP12.6 deficiency in mice facilitates the inducibility of atrial fibrillation.</a> Heart Rhythm. 5, 1047-1047 (2008).</li><li>Chelu, M. G., Sarma, S., Sood, S., Wang, S., van Oort, R. J., Skapura, D. G., Li, N., Santonastasi, M., Muller, F. U., Schmitz, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calmodulin+kinase+II-mediated+sarcoplasmic+reticulum+Ca2++leak+promotes+atrial+fibrillation+in+mice.">Calmodulin kinase II-mediated sarcoplasmic reticulum Ca2+ leak promotes atrial fibrillation in mice.</a> J Clin Invest. 119, 1940-1940 (2009).</li><li>Mathur, N., Subeena, S., Wang, S., van Oort, R. J., Sarma, S., Li, N., Skapura, D., Bayle, J. H., Valderrabano, M., Wehrens, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sudden+Infant+Death+Syndrome+in+Mice+With+an+Inherited+Mutation+in+RyR.">Sudden Infant Death Syndrome in Mice With an Inherited Mutation in RyR.</a> Circ Arrhythmia Electrophysiol. , (2009).</li></ol>

心臓カテーテル検査中に、カテーテル挿入時の広範な出血は、血液量減少に起因する心拍数を増加する可能性があります。このケースでは、滅菌生理食塩水の腹腔内注射（0.3 1.0 mL）を充填圧力を正規化ができ、マウスでの血行動態のストレスを軽減。 イソフルランよりも長い2時間、またはイソフルランの高濃度（&gt; 2％）への曝露がマウスの心臓と呼吸機能を抑制する...

programmed electrical stimulation in mice

遺伝子組み換えマウスは、伝導障害、心房と心室性不整脈、および心臓突然死の分子メカニズムを研究するのが好ましい動物モデルとして浮上している。<sup&gt; 1</sup&gt;心内ペーシング研究は頸静脈に挿入1.1F octapolarカテーテルを使用してマウスで行われ、右心房と心室に進出することができます。ここで、我々は、マウスでプログラムされた電気刺激を行うために必要な手順を示しています。心房の心内ペーシングが外部刺激を用いて行われるのに対し、表面心電図と心内electrogramsは、心房、房室接合部、および心室心筋に同時に記録されます。このように、マウスでプログラムされた電気刺激は伝導欠陥や不整脈の根底にある分子メカニズムを探求するユニークな機会を提供しています。

Watch this Scientific Journal Video about Programmed Electrical Stimulation in Mice at JoVE.com

Programmed Electrical Stimulation in Mice

プログラム電気刺激は、心臓の伝導特性を決定する能力、および様々なペーシングプロトコルを使用して不整脈を誘発し、終了する可能性を提供します。経静脈カテーテルを使用して、心内電位図記録は、不整脈基質を同定するためにプログラムされた電気刺激のプロトコールに従って、マウスで入手できます。

マウスにおけるプログラム電気刺激

Genetically-modified mice have emerged as a preferable animal model to study the molecular mechanisms underlying conduction abnormalities, atrial and ...

programmed-electrical-stimulation-in-mice

Research

JoVE Journal

Medicine

20.3K ビュー.  Baylor College of Medicine (BCM). プログラム電気刺激は、心臓の伝導特性を決定する能力、および様々なペーシングプロトコルを使用して不整脈を誘発し、終了する可能性を提供します。経静脈カテーテルを使用して、心内電位図記録は、不整脈基質を同定するためにプログラムされた電気刺激のプロトコールに従って、マウスで入手できます。

ビデオ: Programmed Electrical Stimulation in Mice

Long-term Potentiation of Perforant Pathway-dentate Gyrus Synapse in Freely Behaving Mice

Studies of long-term potentiation of synaptic efficacy, an activity-dependent synaptic phenomenon having properties that make it attractive as a potential cellular mechanism underlying learning and information storage, have long been used to elucidate the physiology of various neuronal circuits in the hippocampus, amygdala, and other limbic and cortical structures. With this in mind, transgenic mouse models of neurological diseases represent useful platforms to conduct long-term&#160;potentiation (LTP) studies to develop a greater understanding of the role of genes in normal and abnormal synaptic communication in neuronal networks involved in learning, emotion and information processing. This article describes methodologies for reliably inducing LTP in the freely behaving mouse. These methodologies can be used in studies of transgenic and knockout freely&#160;behaving mouse models of neurodegenerative diseases.

Transgenic and knockout mouse models of neurological diseases are useful for studying the role of genes in normal and abnormal neurophysiology. This article describes methodologies which can be used to study long-term potentiation, a cellular mechanism which may underlie learning and memory, in transgenic and knockout freely&#160;behaving mouse models of neuropathology.

自由に行動するマウスにおける有孔質路 - 歯状回シナプスの長期増強

Studies of long-term potentiation of synaptic efficacy, an activity-dependent synaptic phenomenon having properties that make it attractive as a potential ...

行動学

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional capacity. Transcutaneous surface stimulation of skeletal muscle involves a current flow between a cathode and an anode, thereby inducing excitement of the motor unit and the surrounding muscle fibers. 
NMES is an attractive modality to evaluate skeletal muscle adaptive responses for several reasons. First, it provides a reproducible experimental model in which physiological adaptations, such as myofiber hypertophy and muscle strengthening6, angiogenesis7-9, growth factor secretion9-11, and muscle precursor cell activation12 are well documented. Such physiological responses may be carefully titrated using different parameters of stimulation (for Cochrane review, see 13). In addition, NMES recruits motor units non-selectively, and in a spatially fixed and temporally synchronous manner14, offering the advantage of exerting a treatment effect on all fibers, regardless of fiber type. Although there are specified contraindications to NMES in clinical populations, including peripheral venous disorders or malignancy, for example, NMES is safe and feasible, even for those who are ill and/or bedridden and for populations in which rigorous exercise may be challenging. 
Here, we demonstrate the protocol for adapting commercially available electrodes and performing a NMES protocol using a murine model. This animal model has the advantage of utilizing a clinically available device and providing instant feedback regarding positioning of the electrode to elicit the desired muscle contractile effect. For the purpose of this manuscript, we will describe the protocol for muscle stimulation of the anterior compartment muscles of a mouse hindlimb.

A murine model of neuromuscular electrical stimulation (NMES), a safe and inexpensive clinical modality, to the anterior compartment muscles is described. This model has the advantage of modifying a readily available clinical device for the purpose of eliciting targeted and specific muscle contractions in mice.

神経筋電気刺激による筋力トレーニングのマウスモデル

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional ...

医学

Two Different Real-Time Place Preference Paradigms Using Optogenetics within the Ventral Tegmental Area of the Mouse

Understanding how neuronal activation leads to specific behavioral output is fundamental for modern neuroscience. Combining optogenetics in rodents with behavioral testing in validated paradigms allows the measurement of behavioral consequences upon stimulation of distinct neurons in real-time with high spatial and temporal selectivity, and thus the establishment of causal relationships between neuronal activation and behavior. Here, we describe a step-by-step protocol for a real-time place preference (RT-PP) paradigm, a modified version of the classical conditioned place preference (CPP) test. The RT-PP is performed in a three-compartment apparatus and can be utilized to answer if optogenetic stimulation of a specific neuronal population is rewarding or aversive. We also describe an alternative version of the RT-PP protocol, the so-called neutral compartment preference (NCP) protocol, which can be used to confirm aversion. The two approaches are based on extensions of classical methodology originating from behavioral pharmacology and recent implementation of optogenetics within the neuroscience field. Apart from measuring place preference in real time, these setups can also give information regarding conditioned behavior. We provide easy-to-follow step-by-step protocols alongside examples of our own data and discuss important aspects to consider when applying these types of experiments.

Here we present two easy-to-follow step-by-step protocols for place preference paradigms using optogenetics in mice. Using these two different setups, preference and avoidance behaviors can be solidly assessed within the same apparatus with high spatial and temporal selectivity, and in a straightforward manner.

マウスの腹骨テグメンタル領域内の光遺伝学を用いた2つの異なるリアルタイムプレイス優先パラダイム

Understanding how neuronal activation leads to specific behavioral output is fundamental for modern neuroscience. Combining optogenetics in rodents with ...

Optogenetic Manipulation of Neuronal Activity to Modulate Behavior in Freely Moving Mice

Optogenetic modulation of neuronal circuits in freely moving mice affects acute and long-term behavior. This method is able to perform manipulations of single neurons and region-specific transmitter release, up to whole neuronal circuitries in the central nervous system, and allows the direct measurement of behavioral outcomes. Neurons express optogenetic tools via an injection of viral vectors carrying the DNA of choice, such as Channelrhodopsin2 (ChR2). Light is brought into specific brain regions via chronic optical implants that terminate directly above the target region. After two weeks of recovery and proper tool-expression, mice can be repeatedly used for behavioral tests with optogenetic stimulation of the neurons of interest.
Optogenetic modulation has a high temporal and spatial resolution that can be accomplished with high cell specificity, compared to the commonly used methods such as chemical or electrical stimulation. The light does not harm neuronal tissue and can therefore be used for long-term experiments as well as for multiple behavioral experiments in one mouse. The possibilities of optogenetic tools are nearly unlimited and enable the activation or silencing of whole neurons, or even the manipulation of a specific receptor type by light.
The results of such behavioral experiments with integrated optogenetic stimulation directly visualizes changes in behavior caused by the manipulation. The behavior of the same animal without light stimulation as a baseline is a good control for induced changes. This allows a detailed overview of neuronal types or neurotransmitter systems involved in specific behaviors, such as anxiety. The plasticity of neuronal networks can also be investigated in great detail through long-term stimulation or behavioral observations after optical stimulation. Optogenetics will help to enlighten neuronal signaling in several kinds of neurological diseases.

With optogenetic manipulation of specific neuronal populations or brain regions, behavior can be modified with high temporal and spatial resolution in freely moving animals. By using different optogenetic tools in combination with chronically implanted optical fibers, a variety of neuronal modulations and behavioral testing can be performed.

自由に動くマウスの行動を調節する神経活動の光遺伝学的操作

Optogenetic modulation of neuronal circuits in freely moving mice affects acute and long-term behavior. This method is able to perform manipulations of ...

Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) is a promising approach when pharmacological treatment fails or neurosurgery is not recommended. Acute brain slices coupled to microelectrode arrays (MEAs) represent a valuable tool to study neuronal network interactions and their modulation by electrical stimulation. As compared to conventional extracellular recording techniques, they provide the added advantages of a greater number of observation points and a known inter-electrode distance, which allow studying the propagation path and speed of electrophysiological signals. However, tissue oxygenation may be greatly impaired during MEA recording, requiring a high perfusion rate, which comes at the cost of decreased signal-to-noise ratio and higher oscillations in the experimental temperature. Electrical stimulation further stresses the brain tissue, making it difficult to pursue prolonged recording/stimulation epochs. Moreover, electrical modulation of brain slice activity needs to target specific structures/pathways within the brain slice, requiring that electrode mapping be easily and quickly performed live during the experiment. Here, we illustrate how to perform the recording and electrical modulation of 4-aminopyridine (4AP)-induced epileptiform activity in rodent brain slices using planar MEAs. We show that the brain tissue obtained from mice outperforms rat brain tissue and is thus better suited for MEA experiments. This protocol guarantees the generation and maintenance of a stable epileptiform pattern that faithfully reproduces the electrophysiological features observed with conventional field potential recording, persists for several hours, and outlasts sustained electrical stimulation for prolonged epochs. Tissue viability throughout the experiment is achieved thanks to the use of a small-volume custom recording chamber allowing for laminar flow and quick solution exchange even at low (1 mL/min) perfusion rates. Quick MEA mapping for real-time monitoring and selection of stimulating electrodes is performed by a custom graphic user interface (GUI).

We illustrate how to perform recording and electrical modulation of 4-aminopyridine-induced epileptiform activity in rodent brain slices using microelectrode arrays. A custom recording chamber maintains tissue viability throughout prolonged experimental sessions. Live electrode mapping and selection of stimulating pairs are performed by a custom graphical user interface.

記録と電極アレイに結合された齧歯動物の脳スライスにおけるてんかん活動の変調

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) ...

神経科学

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

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings provide a sensitive approach to measure nerve conduction in humans and rodent models. To broaden the technical possibilities for electromyography in mice, the measurement of compound muscle action potentials (CMAPs) from the brachial plexus nerve in the forelimb using needle electrodes is described here. CMAP recordings after stimulating the sciatic nerve in hindlimbs have been previously described. The newly introduced method here allows for the evaluation of the nerve conductivity at an additional site, and thus provides a more profound overview of the neuromuscular functionality. The technique provides information on both the relative number of functional axons and the myelination level. Thereby, this method can be applied to assess both axonal diseases as well as demyelinating conditions. This minimally invasive method does not require extraction of the nerve and therefore it is suitable for repeated measurements for longitudinal follow-up in the same animal. Similar recordings are performed in clinical setups to emphasize the translational relevance of the method.

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

The measurement of nerve conduction is a useful tool to assess mouse models of neurodegeneration but it is frequently only applied to stimulate the sciatic nerve in hindlimbs. Here, we describe a technique to measure compound muscle action potential (CMAP) in vivo in the mouse forelimb muscles innervated by the brachial plexus.

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 ...

An Invasive Method for the Activation of the Mouse Dentate Gyrus by High-frequency Stimulation

Electrical high-frequency stimulation (HFS), using implanted electrodes targeting various brain regions, has been proven as an effective treatment for various neurological and psychiatric disorders. HFS in the deep region of the brain, also named deep-brain stimulation (DBS), is becoming increasingly important in clinical trials. Recent progress in the field of high-frequency DBS (HF-DBS) surgery has begun to spread the possibility of utilizing this invasive technique to other situations, such as treatment for major depression disorder (MDD), obsessive-compulsive disorder (OCD), and so on. 
Despite these expanding indications, the underlying mechanisms of the beneficial effects of HF-DBS remain enigmatic. To address this question, one approach is to use implanted electrodes that sparsely activate distributed subpopulations of neurons by HFS. It has been reported that HFS in the anterior nucleus of the thalamus could be used for the treatment of refractory epilepsy in the clinic. The underlying mechanisms might be related to the increased neurogenesis and altered neuronal activity. Therefore, we are interested in exploring the physiological alterations by the detection of neuronal activity as well as neurogenesis in the mouse dentate gyrus (DG) before and after HFS treatment. 
In this manuscript, we describe methodologies for HFS to target the activation of the DG in mice, directly or indirectly and in an acute or chronic manner. In addition, we describe a detailed protocol for the preparation of brain slices for c-fos and Notch1 immunofluorescent staining to monitor the neuronal activity and signaling activation and for bromodeoxyuridine (BrdU) labeling to determine the neurogenesis after the HF-DBS induction. The activation of the neuronal activity and neurogenesis after the HF-DBS treatment provides direct neurobiological evidence and potential therapeutic benefits. Particularly, this methodology can be modified and applied to target other interested brain regions such as the basal ganglia and subthalamic regions for specific brain disorders in the clinic.

This protocol shows how to set up a reliable HFS method in mice. Neurons throughout the hippocampal dentate gyrus are electrically stimulated by HFS directly and indirectly in vivo. Neuronal activity and molecular signaling are examined by c-fos and Notch1 immunofluorescent staining, respectively; neurogenesis is quantified by bromodeoxyuridine labeling assay.

高周波刺激によるマウスの海馬歯状回の活性化のための侵襲的な方法

Electrical high-frequency stimulation (HFS), using implanted electrodes targeting various brain regions, has been proven as an effective treatment for ...

Transcranial Direct Current Stimulation (tDCS) in Mice

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for several neuropsychiatric diseases. The biological effects of tDCS are not fully understood, which is in part explained due to the difficulty in obtaining human brain tissue. This protocol describes a tDCS mouse model that uses a chronically implanted electrode allowing the study of the long-lasting biological effects of tDCS. In this experimental model, tDCS changes the cortical gene expression and offers a prominent contribution to the understanding of the rationale for its therapeutic use.

Transcranial Direct Current Stimulation tDCS in Mice

Transcranial direct current stimulation (tDCS) is a therapeutic technique proposed to treat psychiatric diseases. An animal model is essential for understanding the specific biological alterations evoked by tDCS. This protocol describes a tDCS mouse model that uses a chronically implanted electrode.

マウスにおける経頭蓋直流電流刺激 (tDCS)

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for ...

脳血流と電場のマッピング:マウスにおける電極配置

Placement of Extracranial Stimulating Electrodes and Measurement of Cerebral Blood Flow and Intracranial Electrical Fields in Anesthetized Mice

The detection of cerebral blood flow (CBF) responses to various forms of neuronal activation is critical for understanding dynamic brain function and variations in the substrate supply to the brain. This paper describes a protocol for measuring CBF responses to transcranial alternating current stimulation (tACS). Dose-response curves are estimated both from the CBF change occurring with tACS (mA) and from the intracranial electric field (mV/mm). We estimate the intracranial electrical field based on the different amplitudes measured by glass microelectrodes within each side of the brain. In this paper, we describe the experimental setup, which involves using either bilateral laser Doppler (LD) probes or laser speckle imaging (LSI) to measure the CBF; as a result, this setup requires anesthesia for the electrode placement and stability. We present a correlation between the CBF response and the current as a function of age, showing a significantly larger response at higher currents (1.5 mA and 2.0 mA) in young control animals (12-14 weeks) compared to older animals (28-32 weeks) (p &lt; 0.005 difference). We also demonstrate a significant CBF response at electrical field strengths &lt;5 mV/mm, which is an important consideration for eventual human studies. These CBF responses are also strongly influenced by the use of anesthesia compared to awake animals, the respiration control (i.e., intubated vs. spontaneous breathing), systemic factors (i.e., CO2), and local conduction within the blood vessels, which is mediated by pericytes and endothelial cells. Likewise, more detailed imaging/recording techniques may limit the field size from the entire brain to only a small region. We describe the use of extracranial electrodes for applying tACS stimulation, including both homemade and commercial electrode designs for rodents, the concurrent measurement of the CBF and intracranial electrical field using bilateral glass DC recording electrodes, and the imaging approaches. We are currently applying these techniques to implement a closed-loop format for augmenting the CBF in animal models of Alzheimer's disease and stroke.

著者スポットライト:アルツハイマー病モデルにおける脳血流を改善するための刺激ベースのアプローチ 

We describe a protocol for assessing dose-response curves for extracranial stimulation in terms of brain electrical field measurements and a relevant biomarker-cerebral blood flow. Since this protocol involves invasive electrode placement into the brain, general anesthesia is needed, with spontaneous breathing preferred rather than controlled respirations.

麻酔マウスにおける頭蓋外刺激電極の配置と脳血流および頭蓋内電界の測定

The detection of cerebral blood flow (CBF) responses to various forms of neuronal activation is critical for understanding dynamic brain function and ...

マウスにおけるプログラム電気刺激

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自由に行動するマウスにおける有孔質路 - 歯状回シナプスの長期増強

神経筋電気刺激による筋力トレーニングのマウスモデル

マウスの腹骨テグメンタル領域内の光遺伝学を用いた2つの異なるリアルタイムプレイス優先パラダイム

自由に動くマウスの行動を調節する神経活動の光遺伝学的操作

記録と電極アレイに結合された齧歯動物の脳スライスにおけるてんかん活動の変調

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

高周波刺激によるマウスの海馬歯状回の活性化のための侵襲的な方法

マウスにおける経頭蓋直流電流刺激 (tDCS)

麻酔マウスにおける頭蓋外刺激電極の配置と脳血流および頭蓋内電界の測定

麻酔マウスにおける頭蓋外刺激電極の配置と脳血流および頭蓋内電界の測定