この作品は、L. Renemanに授与科学研究費オランダ機構（NWO）（Veniない。916.86.125）によって賄われています。資金提供者は、研究デザイン、データ収集と分析、公開することを決定、または原稿の準備で何の役割もありませんでした。利害の衝突はありません。

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我々は、開示することは何もありません。

5-HT phMRI は 、in vivo での動物の神経伝達物質の機能を評価するための有望なツールです。それは機能的なMRイメージングと5-HTの課題に対する脳の反応を可視化する。 MRIは、高空間分解能を持っているので、電離放射線への反復暴露を避けるコントラスト剤または放射性標識分子の注入を必要としないために大きな利点を持っています。この手法は、人間と動物の両方の科目に適用されるため、神経伝達物質システムと精神疾患のトランスレーショナルリサーチのために非常に適しています。そのアプリケーションはもちろん、5-HT経路に限定されるものではなく、既に動物の5,15およびヒト22の両方におけるドーパミン作動薬の効果を評価するために広く使用されています。 すでにマーティンとSibson 11スチュワード20日の記事で指摘したようにそれにもかかわらず、小さな動物のphMRIは、依然として厳しい。これらの課題の一つは、メンテナンスoです画像取得時にf安定した生理的パラメータを設定します。ほとんどの麻酔薬は、心臓血管機能を変化させるとphMRIそれは、任意の血行動態の変化が与えられた薬物のチャレンジにもっぱら起因していることを確認することが不可欠である心血管疾患/血行動態パラメータに決定的に依存していることを考えることができます。それはPco 2のレベルがベースライン取得時に一定に保たれるので、非常に重要です。機械的人工換気は、生理学的安定性を確保することができ、頻繁に実験のこのタイプで使用されています。しかし我々は将来的に長期的な研究を実行する可能性を開いたままに自由呼吸動物を使用することにしました。代わりに、我々は広範囲に安定した血管反応性を維持するために機能的なスキャンの開始前に、この方法で正常範囲内に生理的な安定性を確保するために（そして変更された）呼吸と血液ガスの値を監視し、T2 * / T2信号。脳の血行動態とmetabolisに関する一般的な麻酔薬の影響についての文献mは20豊富で、この原稿の範囲を超えています。吸入麻酔薬で、麻酔の深さは、迅速かつ容易に制御することができるので、我々は、この特定のプロトコルのイソフルラン±2％でガス麻酔を使うことにしました。これは、画像取り込みの開始前に正常範囲に安定PCO 2のレベルを確保するために我々のセットアップで重要である。イソフルランは、今日最も一般的に使用される吸入麻酔薬であり、縦断的研究のために重要である迅速な誘導と回復が可能になります。また、最小の心血管系および呼吸抑制を生成し、骨格筋の良いリラクゼーションを誘発する。 第二に、挑戦的な薬剤の静脈内投与は、ヒトのより小さい動物のより複雑です。大腿動脈と静脈のカニュレーションに必要とされる手術はよく訓練され経験豊富なスタッフを必要とします。それは現時点では、端末の手順で主に使用されるこれらの侵襲的処置のために。しかし、血液の恒常性と尾静脈注射の非侵襲的モニタリングは、縦断的研究23を使用することができる。 さらに、動物phMRIのために固有のものではありませんテクニックにはいくつかのより一般的な制限があります。マーティンとSibson 11で指摘したようにさらに、すべてのfMRIの研究の潜在的な混乱させる、それが挑戦によって誘発される脳活動の変化が神経活動の変化はなく、周辺全身への影響を反映していると想定されるということです。特に深い脳の構造で、神経血管カップリングの比較的貧しい人々の理解は（神経活動の変化と血行動態の変化との関係）のまま。皮質で神経血管カップリングを決定するためにLogothetis 10によって行わ種の研究は、まだ脳の他の部分で実施されていない。それが重要な構造のBOLD信号の増加などの線条体や扁桃体はテリン何であるかは不明であるので、ニューロンの活動について教えてG。我々はこの時点で言うことができる最高の脳の領域が与えられた課題に反応し、治療および/または条件に応じて、我々は脳の反応性の有意な変化を監視することができますということです。これは、主にMRIデータと生理学的応答の両方を見て確認することができます。脳の活性化の一般的なパターンは、この場合、高い5-HT神経支配ではなく、できるだけ一般的な血管の応答で、ある領域に領域を特定し、制限する必要があります。また、血管と血行動態の変化の間に別の一時的なプロファイルが期待されています。血圧の変化は数分以内にベースライン値に戻るのに対し、BOLD活性化に対する薬剤の効果は画像の取り込みが終了するまで表示されてフルオキセチンの場合ですと、この薬の知っている薬物動態学的特性に対応しています。最後に、すべての動物の生理学的応答は、被験者間の比較を行うために、似ている必要があります。 NonetheleSS、それは5-HTによる局所血流の神経性調節4が存在することが知られている。したがって、それはBOLD信号の局所的変化は血管の近傍に5-HTの放出による血管の変化に帰することを除外することはできません。これらの効果は、ローカルニューロンの活性化に関連付けられていないため、偽陽性の結果とみなすことができるが、それはまた5-HTシステムの全体的な特定の機能（これも3を参照）のインデックスです。 この手法の重要なステップは、広範囲の生理学的反応を監視するために、動物の生理的条件下では、画像取得前との間に安定していることを確認してくださいすることがあります。また、スキャナの条件は、できるだけ安定しており、各動物のために正確に同じである必要があります。シーケンスの信号安定性をチェックして、実験の開始前に確認する必要があります。さらに、小さな被写体grouで、常に十分な大きさの統計的検出力を持っていることを確認してくださいpsの。一般に動物phMRIの実験的検討事項の素敵なレビューのために、スチュワード20を参照し、ラットとマウスにおける薬理fMRIのための実験プロトコルの追加例えば、フェラーリ5を参照してください。 ここで説明する手法の可能な変更はたくさんあり​​ます。一つは、可能性： <ol><li>薬物作用6,19の基礎となるメカニズムを明らかにするためにそのような別のSSRIまたは5-HT受容体（ANT）アゴニスト16,13,18,7、あるいは二重の課題として5-HTの挑戦のために別の薬剤を使用して; </li><li>そのような別の麻酔体制、機械的人工換気、血液プール造影剤の代わりに、BOLD 15、縦断的研究（動物が生きて維持する必要があるなど、さまざまな実験のセットアップを使用するので、全く観血式血圧/血液ガス測定しない、および/ ​​または機械的人工換気MR-互換性のあるELEを使用して、このような神経活動の記録として、他の（侵襲性）メソッドとの組み合わせでも）が可能であるか、またはctrodes 10またはPET / SPECT研究4; </li><li> &quot;P-ブロック&quot;マッキー12に記載の方法または機能の接続性の分析15などの異なるMRIデータ分析方法を使用します。 </li></ol>あなたが実験で行った選択の可能性および/またはあなたのラボとあなたが答えたい研究課題のタイプ内の経験に大きく依存している。

<table border="1"><tbody><tr><td> 試薬/装置の名前 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td>イソフルレン</td><td> ABBOTT <a href="http://www.abbott.co.uk/" target="_blank">アボット·ラボラトリーズ（株）、英国メイデンヘッド</a> </td><td>いいえB506ません</td><td>医療用空気と混合</td></tr><tr><td>医療用空気</td><td> <a href="http://www.bochealthcare.co.uk/" target="_blank">BOCヘルスケア</a> </td><td></td><td></td></tr><tr><td>加温パッド</td><td> <a href="http://www.harvardapparatus.co.uk/" target="_blank">ハーバード装置</a> </td><td> 507223F <a href="http://www.harvardapparatus.co.uk/webapp/wcs/stores/servlet/haisku3_10001_11555_39108_-1_HAUK_ProductDetail_N_37610_37611_37613" target="_blank">フレキシブルプローブ、ミディアム、230 VAC、50 Hzの完全な恒温動物ブランケットシステム</a> </td><td></td></tr><tr><td>シルクリットル<html> igature </td><td> <a href="http://www.harvardapparatus.com/" target="_blank">http://www.harvardapparatus.com/</a> </td><td> <a href="http://www.harvardapparatus.com/webapp/wcs/stores/servlet/product_11051_10001_43051_-1_HAI_ProductDetail" target="_blank">非吸収性2から0黒編みシルク</a> 猫のNum 51から7631 </td><td></td></tr><tr><td> PE-50カニューレ</td><td> <a href="http://www.scientificlabs.co.uk/" target="_blank">http://www.scientificlabs.co.uk/~~V</a> </td><td> <a href="http://www.scientificlabs.co.uk/product/TUB3662" target="_blank">PortexチューブPE 0.58x0.96mm</a> </td><td> 0.58 ID 0.96外径mm </td></tr><tr><td>ヘパリンナトリウム</td><td> <a href="http://www.leo-pharma.com/Home/Products/List-of-LEO-Pharma-products.aspx" target="_blank">レオの研究所株式会社、バックス、イギリス</a> </td><td>ヘパリンナトリウム1000IU/ml </td><td> 15 U / mlの</td></tr><tr><td> Vetbond組織接着剤</td><td> <a href="http://solutions.3m.co.uk" target="_blank">3M</a> </td><td>TTP :/ / solutions.3m.co.uk/wps/portal/3M/en_GB/HealthCare/Home/ProdInfo/VetProducts / &quot;ターゲット=&quot; _blank &quot;&gt; M Vetbondの組織接着剤</td><td></td></tr><tr><td>監視システム</td><td> <a href="http://www.i4sa.com/" target="_blank">SA·インスツルメンツ</a> </td><td> <a href="http://www.i4sa.com" target="_blank">http://www.i4sa.com</a> <a href="http://69.20.42.253/web_app/main/defaultProduct.aspx?ID=16&amp;PT=3" target="_blank">モデル1025L監視システム</a> </td><td>モニター呼吸と温度</td></tr><tr><td>圧力トランスデューサー</td><td> <a href="http://www.biopac.com/" target="_blank">モンテシステム株式会社</a> </td><td> <a href="http://www.biopac.com/blood-pressure-transducer" target="_blank">血圧トランスデューサ- TSD104A</a> <a href="http://www.biopac.com/data-acquisition-analysis-system-mp150-system-windows" target="_blank">MP150データ·アクイジション·システム- WIN - MP150WSW</a> </td><td>モニター血圧</td></tr><tr><td> RapidLab血液ガス分析装置</td><td> <a href="http://www.medical.siemens.com/" target="_blank">シーメンスの診断</a> </td><td> <a href="http://www.medical.siemens.com/webapp/wcs/stores/servlet/ProductDisplay~q_catalogId~e_-111~a_catTree~e_100001,1023069,1016115~a_langId~e_-111~a_productId~e_172940~a_storeId~e_10001.htm" target="_blank">RAPIDLab 348分の248システム</a> </td><td></td></tr><tr><td> 4.7T動物スキャナ</td><td>アジレント·テクノロジー（以前MAGNEX） 4.7T周波数199.845 MHzの</td><td></td><td></td></tr><tr><td> MR互換性のある定位ベッド</td><td> <a href="http://www.m2mimaging.com/products/preclinical.html" target="_blank">M2Mイメージング株式会社</a> </td><td>ラットベッド：PA Multi要素50-72-1003/100 AHS </td><td></td></tr><tr><td>コイル</td><td> <a href="http://www.m2mimaging.com/products/preclinical.html" target="_blank">M2Mイメージング株式会社</a> </td><td>ボリュームTH / Rxの RQD1 112分の72 200 </td><td></td></tr><tr><td>塩酸フルオキセチン</td><td> Sigma-Aldrich社</td><td> F-132 </td><td>生理食塩水に5mg/kgを</td></tr></tbody></table>

the use of pharmacological-challenge fmri in pre-clinical research: application to the 5-ht system

薬理学的MRI（phMRI）は、最終的に、うつ病やADHDなどの薬物作用と神経伝達物質関連障害の背後にある基本的な神経生物学的メカニズムを解明するために使用することができ、脳の機能上の物質の影響を研究するための新しい有望な方法です。撮像法（PET、SPECT、CT）の大半と同様、​​それは脳障害の調査と全体的な神経接続の尊重と非侵襲的な方法で神経伝達経路の関連する関数の進捗状況を表しています。また、それはまた、臨床試験への変換のための理想的なツールを提供します。 MRIは、まだ背後にある分子イメージング戦略のPETやSPECTと比較して、しばらく大きな利点は、それによって電離放射線への反復暴露を避けて、高空間分解能やコントラストエージェントまたは無線標識分子の注入の必要はありませんを持っている必要があります。それはgである機能的MRI（fMRI）が広く、研究と臨床の現場で使用されているenerally精神運動タスクを組み合わせる。 phMRIは、fMRIは、特定の困難な薬物の投与を経由してアクティベーション後に生理学的または病理学的条件下では、セロトニンなどの特定の神経伝達物質システム、（5-HT）の調査を可能に適応したものです。 ここで説明する方法の目的は、自由呼吸動物の脳5-HT機能を評価することである。同時に時間の経過とともに機能的なMR画像を取得中に5-HTシステムに挑戦することによって、この課題に対する脳の反応を可視化することができます。動物のいくつかの研究は既に例えば5-HT（剤、選択的再取り込み遮断薬などを放出する）の細胞外濃度の薬剤誘発性の増加が原因で血中酸素レベル依存（BOLD）MRI信号（信号の地域固有の変更を呼び起こすことが示されている酸素とgを供給する血液の供給の増加を介して脳の活性化時に発生する酸化/脱酸素化ヘモグロビン濃度の変化に神経伝達物質の機能の指標を提供する）を要求ニューロンにlucose。また、これらの効果は減少し5-HTの可用性16,13,18,7その治療によって逆にすることができることが示されている。成体ラットでは、急性SSRI投与後のBOLD信号変化は、いくつかの5-HTに関連する脳の領域、すなわち皮質、海馬、視床下部と視床9,16,15に記載されている。 5-HT系の刺激と、この課題への応答は、このように動物とヒトの2,11の両方でその機能の尺度として使用することができます。

Watch this Scientific Journal Video about The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System at JoVE.com

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

この手法の目的は、セロトニン（5-HT）の薬理学的磁気共鳴イメージング（phMRI）と選択的セロトニン再取り込み阻害薬（SSRI）、フルオキセチンと静脈内チャレンジライブと自由呼吸動物の神経伝達物質の機能を評価することである。

前臨床研究における薬理学的チャレンジのfMRIの利用：5-HTシステムへの応用

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

the-use-pharmacological-challenge-fmri-pre-clinical-research

Research

JoVE Journal

Medicine

15.3K ビュー.  Academic Medical Center Amsterdam. この手法の目的は、セロトニン（5-HT）の薬理学的磁気共鳴イメージング（phMRI）と選択的セロトニン再取り込み阻害薬（SSRI）、フルオキセチンと静脈内チャレンジライブと自由呼吸動物の神経伝達物質の機能を評価することである。

ビデオ: The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

新しいハーブ治療法を評価するために敗血症の動物モデルの使用

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

医学

Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact. For this strategy to be successful, recent studies have shown that involvement of the host immune system is essential. Therefore, oncolytic virotherapy should be evaluated within the context of an immunocompetent model. Furthermore, the study of antitumor therapies in tolerized animal models may better recapitulate results seen in clinical trials. Cotton rats, commonly used to study respiratory viruses, are an attractive model to study oncolytic virotherapy as syngeneic models of mammary carcinoma and osteosarcoma are well established. However, there is a lack of published information on the proper handling procedure for these highly excitable rodents. The handling and capture approach outlined minimizes animal stress to facilitate experimentation. This technique hinges upon the ability of the researcher to keep calm during handling and perform procedures in a timely fashion. Finally, we describe how to prepare cotton rat mammary tumor cells for consistent subcutaneous tumor formation, and how to perform intratumoral and intraperitoneal injections. These methods can be applied to a wide range of studies furthering the development of the cotton rat as a relevant pre-clinical model to study antitumor therapy.

Cotton rats are extremely excitable and have a strong flight-or-fight response. A handling method optimized to reduce the stress of the animals is described which will make cotton rats more accessible as a preclinical model.

腫瘍溶解性ウイルスの前臨床評価のための研究のコットンラットの取扱い

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact.  For this strategy to be ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

ヒト前立腺癌の前臨床同所性マウスモデル

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Use of a Low-flow Digital Anesthesia System for Mice and Rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers utilize a syringe pump to directly administer volatile anesthetics into a gas stream. Unlike a traditional vaporizer, it can be used at very low flow rates, making it ideal for use on mice and rats. 
The equipment's capability to use low flow rates could result in a substantial cost savings due to the reduced need for anesthetic agents, compressed gas, and charcoal scavenging filters1. A lower flow rate means less waste of anesthetic gas and likely reduces the risk of anesthetic exposure to laboratory personnel. Thus, the high levels of precision and safety associated with direct injection vaporizers, along with a reduced need for anesthetic agents, compressed gas, and charcoal filters are beneficial for research requiring small animal anesthesia. 
The goal of this protocol is to demonstrate the use of a syringe-driven direct injection vaporizer as part of a digital, low-flow anesthesia system. The direct injection vaporizer is capable of accurately delivering anesthesia at very low flow rates compared to a traditional vaporizer, making it a promising alternative for controlled gas anesthetic delivery to rodents.

Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer.

マウスやラットのための低フローデジタル麻酔システムの利用

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers ...

Ultrahigh Resolution Mouse Optical Coherence Tomography to Aid Intraocular Injection in Retinal Gene Therapy Research

HR-SD-OCT is utilized to monitor the progression of photoreceptor degeneration in live mouse models, assess the delivery of therapeutic agents into the subretinal space, and to evaluate toxicity and efficacy in vivo. HR-SD-OCT uses near infrared light (800-880 nm) and has optics specifically designed for the unique optics of the mouse eye with sub-2-micron axial resolution. Transgenic mouse models of outer retinal (photoreceptor) degeneration and controls were imaged to assess the disease progression. Pulled glass microneedles were used to deliver sub retinal injections of adeno-associated virus (AAV) or nanoparticles (NP) via a trans-scleral and trans-choroidal approach. Careful positioning of the needle into the subretinal space was required prior to a calibrated pressure injection, which delivers fluid into the sub retinal space. Real time subretinal surgery was conducted on our retinal imaging system (RIS). HR-SD-OCT demonstrated progressive uniform retinal degeneration due to expression of a toxic mutant human mutant rhodopsin (P347S) (RHOP347S) transgene in mice. HR-SD-OCT allows rigorous quantification of all the retinal layers. Outer nuclear layer (ONL) thickness and photoreceptor outer segment length (OSL) measurements correlate with photoreceptor vitality, degeneration, or rescue. The RIS delivery system allows real-time visualization of subretinal injections in neonatal (~P10-14) or adult mice, and HR-SD-OCT immediately determines success of delivery and maps areal extent. HR-SD-OCT is a powerful tool that can evaluate the success of subretinal surgery in mice, in addition to measuring vitality of photoreceptors in vivo. HR-SD-OCT can also be used to identify uniform animal cohorts to evaluate the extent of retinal degeneration, toxicity, and therapeutic rescue in preclinical gene therapy research studies.

Here we demonstrate a novel approach to using high resolution spectral-domain optical coherence tomography (HR-SD-OCT) to assist delivery of gene therapy agents into the subretinal space, assess its areal coverage, and characterize photoreceptor vitality.

超高解像度マウス光コヒーレンストモグラフィー網膜遺伝子治療研究における眼内注入を支援するために

HR-SD-OCT is utilized to monitor the progression of photoreceptor degeneration in live mouse models, assess the delivery of therapeutic agents into the ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

オルソトピック心臓移植の前臨床ブタモデル

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Pre-clinical Model of Cardiac Donation after Circulatory Death

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after circulatory death (DCD) is a solution to address this limited availability, but due to a period of prolonged warm ischemia and the risk of tissue injury, its routine use in cardiac transplantation is seldom seen. In this manuscript we provide a detailed protocol closely mimicking current clinical practices in the context of DCD with continuous monitoring of heart function, allowing for the evaluation of novel cardioprotective strategies and interventions to decrease ischemia-reperfusion injury.
In this model, the DCD protocol is initiated in anesthetized Lewis rats by stopping ventilation to induce circulatory death. When systolic blood pressure drops below 30 mmHg, the warm ischemic time is initiated. After a pre-set warm ischemic period, hearts are flushed with a normothermic cardioplegic solution, procured, and mounted onto a Langendorff ex vivo heart perfusion system. Following 10 min of initial reperfusion and stabilization, cardiac reconditioning is continuously evaluated for 60 min using intraventricular pressure monitoring. A heart injury is assessed by measuring cardiac troponin T and the infarct size is quantified by histological staining. The warm ischemic time can be modulated and tailored to develop the desired amount of structural and functional damage. This simple protocol allows for the evaluation of different cardioprotective conditioning strategies introduced at the moment of cardioplegia, initial reperfusion and/or during ex vivo perfusion. Findings obtained from this protocol can be reproduced in large models, facilitating clinical translation.

This protocol shows a simple and flexible approach for the evaluation of new conditioning agents or strategies to increase the feasibility of cardiac donation after circulatory death.

循環死後の心臓寄付の前臨床モデル

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. However, the evaluation of right ventricular pressure (RVP) and pulmonary artery pressure (PAP) remains technically challenging in mice. RVP and PAP are more difficult to measure than left ventricular pressure because of the anatomical differences between the left and right heart systems. In this paper, we describe a stable right heart hemodynamic measurement method and its validation using healthy and PAH mice. This method is based on open-chest surgery and mechanical ventilation support. It is a complicated procedure compared to closed chest procedures. While a well-trained surgeon is required for this surgery, the advantage of this procedure is that it can generate both RVP and PAP parameters at the same time, so it is a preferable procedure for the evaluation of PAH models.

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

マウスにおける右心室系と低酸素誘発性肺動脈高血圧に対する侵襲的血行力学的評価

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

再建微小外科における虚血再灌流損傷の研究のための前臨床ラットモデル

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare recommendations, animals are warmed and monitored during procedures requiring anesthesia. Common anesthesia and physiological monitoring equipment include gas tanks, anesthetic vaporizers and stands, warming controllers and pads, mechanical ventilators, and pulse oximeters. A computer is also necessary for data collection and to run equipment software. In smaller spaces or when performing field work, it can be challenging to configure all this equipment in limited space.
The goal of this protocol is to demonstrate best practices for use of a low-flow digital vaporizer using both compressed oxygen and room air, along with an integrated mechanical ventilator, pulse oximeter, and far infrared warming as an all-inclusive anesthesia and physiological monitoring suite ideal for rodents.

Here, we present a protocol to safely and effectively administer anesthetic gas to mice using a digital, low flow anesthesia system with integrated ventilator and physiological monitoring modules.

げっ歯類のための統合された低流量麻酔気化器、換気装置および生理学的モニタリングシステムの使用

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare ...