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ことができます。したがって、すべての試験は2時間以内で完了することをお勧めします。また、体温が常に正常範囲37.0 ± 1.0℃以内に維持されることが不可欠である低体温と高体温の両方が、心臓のリズムと不整脈基質の存在する可能性に影響を与えます。 各実験は、心房捕捉しきい値を決定することによって開始する必要があります。心房ペーシング、心房閾値、SNRT、AERP、およびAVNERPの場合には洞結節、房室結節、および心房組織の伝導特性が正常であるかどうかを評価するために決定されるべきである。

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

Nanyang Technological University

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

Transthoracic Echocardiography in Mice

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

マウスにおける経胸壁心エコー

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

医学

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 ...

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

マウスで制御子宮裂傷の傷害

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

Breathing-controlled Electrical Stimulation (BreEStim) for Management of Neuropathic Pain and Spasticity

Electrical stimulation (EStim) refers to the application of electrical current to muscles or nerves in order to achieve functional and therapeutic goals. It has been extensively used in various clinical settings. Based upon recent discoveries related to the systemic effects of voluntary breathing and intrinsic physiological interactions among systems during voluntary breathing, a new EStim protocol, Breathing-controlled Electrical Stimulation (BreEStim), has been developed to augment the effects of electrical stimulation. In BreEStim, a single-pulse electrical stimulus is triggered and delivered to the target area when the airflow rate of an isolated voluntary inspiration reaches the threshold. BreEStim integrates intrinsic physiological interactions that are activated during voluntary breathing and has demonstrated excellent clinical efficacy. Two representative applications of BreEStim are reported with detailed protocols: management of post-stroke finger flexor spasticity and neuropathic pain in spinal cord injury.

Breathing-controlled Electrical Stimulation BreEStim for Management of Neuropathic Pain and Spasticity

The purpose is to present a new method, breathing-control electrical stimulation (BreEStim) for management of neuropathic pain and spasticity.

神経因性疼痛、痙縮の管理のための呼吸制御型電気刺激（BreEStim）

Electrical stimulation (EStim) refers to the application of electrical current to muscles or nerves in order to achieve functional and therapeutic goals. ...

Parabiosis in Mice: A Detailed Protocol

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of microvasculature at the site of inflammation. Parabiotic partners share their circulating antigens and thus are free of adverse immune reaction. First described by Paul Bert in 18641, the parabiosis surgery was refined by Bunster and Meyer in 1933 to improve animal survival2. In the current protocol, two mice are surgically joined following a modification of the Bunster and Meyer technique. Animals are connected through the elbow and knee joints followed by attachment of the skin allowing firm support that prevents strain on the sutured skin. Herein, we describe in detail the parabiotic joining of a ubiquitous GFP expressing mouse to a wild type (WT) mouse. Two weeks after the procedure, the pair is separated and GFP positive cells can be detected by flow cytometric analysis in the blood circulation of the WT mouse. The blood chimerism allows one to examine the contribution of the circulating cells from one animal in the other.

Parabiotic joining of two organisms leads to the development of a shared circulatory system. In this protocol, we describe the surgical steps to form a parabiotic connection between a wild-type mouse and a constitutive GFP-expressing mouse.

マウスの並体結合：詳細なプロトコル

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

マウスにおけるタキサンの薬物動態学的および薬力学的分析のための頸動脈煎茶

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

マウスにおける異子宮頸心臓移植

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

A Multimodal Imaging- and Stimulation-based Method of Evaluating Connectivity-related Brain Excitability in Patients with Epilepsy

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations over time in the blood-oxygenation level dependent signal. rs-fcMRI has been applied extensively to identify abnormalities in brain connectivity in different neurologic and psychiatric diseases. However, the relationship among rs-fcMRI connectivity abnormalities, brain electrophysiology and disease state is unknown, in part because the causal significance of alterations in functional connectivity in disease pathophysiology has not been established. Transcranial Magnetic Stimulation (TMS) is a technique that uses electromagnetic induction to noninvasively produce focal changes in cortical activity. When combined with electroencephalography (EEG), TMS can be used to assess the brain's response to external perturbations. Here we provide a protocol for combining rs-fcMRI, TMS and EEG to assess the physiologic significance of alterations in functional connectivity in patients with neuropsychiatric disease. We provide representative results from a previously published study in which rs-fcMRI was used to identify regions with abnormal connectivity in patients with epilepsy due to a malformation of cortical development, periventricular nodular heterotopia (PNH). Stimulation in patients with epilepsy resulted in abnormal TMS-evoked EEG activity relative to stimulation of the same sites in matched healthy control patients, with an abnormal increase in the late component of the TMS-evoked potential, consistent with cortical hyperexcitability. This abnormality was specific to regions with abnormal resting-state functional connectivity. Electrical source analysis in a subject with previously recorded seizures demonstrated that the origin of the abnormal TMS-evoked activity co-localized with the seizure-onset zone, suggesting the presence of an epileptogenic circuit. These results demonstrate how rs-fcMRI, TMS and EEG can be utilized together to identify and understand the physiological significance of abnormal brain connectivity in human diseases.

Resting-state functional-connectivity MRI has identified abnormalities in patients with a wide range of neuropsychiatric disorders, including epilepsy due to malformations of cortical development. Transcranial Magnetic Stimulation in combination with EEG can demonstrate that patients with epilepsy have cortical hyperexcitability in regions with abnormal connectivity.

マルチモーダルImaging-とてんかんの患者で接続関連の脳の興奮性を評価するの刺激に基づく方法

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations ...

Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for erectile dysfunction. However, the methods used vary between laboratories, and pitfalls still exist. The goal of this study was to describe a surgical technique that would provide a reliable and reproducible model. By exposing the ischiocavernosus muscle at its point of insertion on the ischial tuberosity, the penile crus could be cannulated with minimal dissection and injury to the structures involved in erectile function. Repeated stimulation of the CN, without the need for lifting and drying, was achieved by using a 125 &#181;m bipolar silver electrode and biocompatible silicon glue to isolate the electrode-nerve complex. This method prevents neuropraxia by reducing stretching and drying the nerve and provides complete isolation of the nerve, negating electrical leakage and preventing stimulation of alternative pathways.

This study describes a simplified surgical procedure and technique for performing cavernous nerve stimulation with the isolation of the nerve-electrode complex using silicone glue and intracavernous pressure measurement.

海綿神経刺激とラットの海綿静脈洞部の圧力の記録

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for ...

Updated Technique for Reliable, Easy, and Tolerated Transcranial Electrical Stimulation Including Transcranial Direct Current Stimulation

Transcranial direct current stimulation (tDCS) is a noninvasive method of neuromodulation using low-intensity direct electrical currents. This method of brain stimulation presents several potential advantages compared to other techniques, as it is noninvasive, cost-effective, broadly deployable, and well-tolerated provided proper equipment and protocols are administered. Even though tDCS is apparently simple to perform, correct administration of the tDCS session, especially the electrode positioning and preparation, is vital for ensuring reproducibility and tolerability. The electrode positioning and preparation steps are traditionally also the most time consuming and error-prone. To address these challenges, modern tDCS techniques, using fixed-position headgear and pre-assembled sponge electrodes, reduce complexity and setup time while also ensuring that the electrodes are consistently placed as intended. These modern tDCS methods present advantages for research, clinic, and remote-supervised (at home) settings. This article provides a comprehensive step-by-step guide for administering a tDCS session using fixed-position headgear and pre-assembled sponge electrodes. This guide demonstrates tDCS using commonly applied montages intended for motor cortex and dorsolateral prefrontal cortex (DLPFC) stimulation. As described, selection of the head size and montage-specific headgear automates electrode positioning. Fully assembled pre-saturated snap-electrodes are simply affixed to the set position snap-connectors on the headgear. The modern tDCS method is shown to reduce setup time and reduce errors for both novice and expert operators. The methods outlined in this article can be adapted to different applications of tDCS as well as other forms of transcranial electrical stimulation (tES) such as transcranial alternating current stimulation (tACS) and transcranial random noise stimulation (tRNS). However, since tES is application specific, as appropriate, any methods recipe is customized to accommodate subject, indication, environment, and outcome specific features.

When administering transcranial direct current stimulation (tDCS), reproducible electrode preparation and placement are vital for a tolerated and effective session. The purpose of this article is to demonstrate updated modern setup procedures for the administration of tDCS and related transcranial electrical stimulation techniques, such as transcranial alternating current stimulation (tACS).

経頭蓋直流刺激を含む信頼性、容易、および許容経頭蓋電気刺激のための更新された技術

Transcranial direct current stimulation (tDCS) is a noninvasive method of neuromodulation using low-intensity direct electrical currents. This method of ...