この作品は、国立衛生研究所（R01DK58096ヴィンセントPoitoutまで）によってサポートされていました。ヴィンセントPoitoutは、糖尿病と膵β細胞機能におけるカナダリサーチチェアを保持しています。ベイダーZarroukiはメルクとイーライリリーからポスドクフェローシップを受けた。 GhislaineFontésは、カナダ糖尿病協会からのポスドクフェローシップによってサポートされていました。

概要：; 7日間の回復期間を許可する、スイベルやケージ内を自由に移動するために動物を可能にするカウンターウェイトシステムを使用してポンプにカテーテルを接続する手順は、全身麻酔下に右頸静脈と左頸動脈をcatheterizingで構成されています; 72時間グルコース及び/又はイントラリピッドを（ヘパリン9を注入したときに約80％unsaturated/20％飽和脂肪酸の混合物を生成する大豆油エマルジ ​​ョン）注入する。 1。右頚静脈と左頸動脈のCanulation <ol><li>外科用器具を滅菌する。 Canulationチューブも手続き前に液体滅菌剤（2.6％グルタルアルデヒド）を使用して冷滅菌でなければなりません。 16-24時間、オートクレーブ容器にチューブを浸します。すすぎ、使用前にグルタルアルデヒドのすべてのトレースを削除するには、滅菌蒸留水で十分に洗い流してください。 </li><li>計算するためにラットを秤量薬の投与量： カルプロフェン5 mg / kg体重：希釈、1/10 =本体重量（g）×0.001ミリリットルSC（鎮痛剤） 希釈の1/10 = BW（G）X 0.0012ミリリットルSC（抗コリン）：0.01 mg / kgでのグリコ </li><li>イソフルランと酸素を使用してラットを麻酔。 </li><li>その胃にラットを置きます。肩のベースに耳の後ろの領域を剃る。その背中にネズミを置き。前足に首下の領域を剃る。 </li><li>プレップクロルヘキシジン、アルコール、ヨウ素と手術部位。薬を管理します。 </li><li>手術野にラットを転送します。 </li><li>無菌技術を使用して、ヘパリン化生理食塩水の5 Uでいっぱい、1mlのシリンジに接続されているPE-50カニューレと右頚静脈と左頸動脈にカニューレを挿入する。回復期間中に凝固を避けるためにヘパリン加生理食塩水を50 Uとカニューレをフラッシュします。鈍化23G針を使用してください。 23Gピンでそれぞれカニューレを閉じます。 </li><li>手術後、オフ約2.5ミリメートルトリムボトム門歯と食べられているからカニューレを防ぐために、点滴のジャケットにラットを配置。 </li><li>イソフルラン避難支援する酸素（1L / 10分間分）を提供します。 </li><li>それは完全に目を覚ましになるまで加熱したケージにラットを置きます。 </li><li>注入条件ごとに1つを使用するために4匹のラット（ 表1）を操作してください。 </li></ol> 2。術後のケア（カテーテルの後の外科的治療と接続） <ol><li> 1日目と2日目手術後にラットの重量を量る。 </li><li> 1日目手術後に、一度2日目手術後に二回グリコ（BW（G）×0.00048ミリリットル）SCを管理します。 </li><li>必要に応じて追加の支援的な治療を投与することができる：流体、加熱パッド、ぬれたダイエット、酸素療法、鎮痛薬、抗コリン薬。 </li><li> 7日目手術後では、注入のための流量を計算するためにラットの重量を量る。 </li><li>ケージグリル上部（図1）に搭載されたテザーとスイベルを使用して注入システムに各ラットを接続します。</li><li>血栓を除去するためにヘパリン化生理食塩水の5 Uとカニューレをフラッシュします。凝固防ぐためにヘパリン加生理食塩水の50 Uでもう一度カニューレをフラッシュします。 </li><li>ラットは注入を開始する前に、少なくとも24時間、テザーとスイベルに順応することができます。 </li></ol> 3。注入<ol><li>各ラットと対策血糖の頸動脈からの血液0.15ミリリットルを描画します。頸静脈カニューレをフラッシュします。各ラットの両方のカニューレ内凝固を防ぐためにヘパリン化生理食塩水の50 Uを使用してください。 </li><li> 2％EDTAを含有するの0.5ml収集チューブに血液サンプルを移す。 2分間10,000 rpmで遠心し、-20℃で血漿を凍結</li><li>下記の注入条件ごとに2つの60ミリリットル注射器を充填する。ポンプの後ろの位置にポンプと場所シリンジ2の正面の位置に置きシリンジ1。 </li><li> Y-コネクタと滅菌されたCO-EX T22チューブと一緒にソリューションの各ペアに参加します。ハーバード33に注射器を置きデュアルシリンジポンプ。 </li><li>ケージ底を変更して、ケージのグリル上から全ての食品を除去します。 </li><li>ケージのグリル上部に標準餌と場所150gの重量を量る。 </li><li>ケージグリル上のスイベルに注射器を接続します。ラインから閉じ込められた空気を除去するために適切に注射器をフラッシュします。 </li><li>輸液システムにラットを接続する前に撮影された体重を使用して、注入流量を計算する。料金はミリリットル/ hの中にグルコース注入率（GIR）を変換し、Microsoft Excelファイルで計算されています。 </li><li>メーカーの設定に応じて60ミリリットル注射器のための流量を管理するためのポンプを設定します。シリンジ1のレート（フロントシリンジ）と注射器2（バックシリンジ）のレートを入力します。 </li><li>ポンプを始動。 </li></ol> 4。モニタリング<ol><li>ポンプを起動した後、そこにはスイベルから、あるいはカニューレから漏れていると、その輸液チューブがねじれていないことを確認していません。また内に気泡が存在しないことを確認チューブ。 </li><li>注入の3時間後に、血糖をモニターするために血液サンプルを取る。 48、24、6の後に繰り返して、注入の72時間。予防策として、血糖はまた、注入の30、34、54、及び60時間後に監視される。血中グルコースは、携帯型グルコース·モニターを使用して、全血の一滴を用いて測定することができる。これは、注入時に採取した血液の量を制限し、したがって有意に変化させる血液量および/またはヘマトクリットを回避する。 </li><li>シリンジ1のレートは、220〜250ミリグラム/ dlとで血糖値を維持するために血糖値に基づいて修正される。遊離脂肪酸を1モル/ Lに維持されるため、シリンジ2のレートは、注入の72時間中に変化しない</li><li>注入条件は、対照動物のための受信されたボリュームは、試験動物（ 表1）受信された体積に相当するように対になっている。 </li><li>注入の24時間後、ケージ底を変更して、ケージのグリルに残っている食べ物を重量を量る。食べ残しの部分tを返しますケージグリルÔ。 48時間後に繰り返します。 </li><li>注入の72時間の間に、必要に応じて毎日の詰め替え注射器。 </li><li>注入時には、常に炎症や不快感の兆候のためラットを観察します。必要に応じて適切な介護を管理します。 </li></ol> 5。注入後の安楽死<ol><li class="jove_content">注入の72時間後、ラットをケタミン/キシラジンカクテル（ケタミンおよびキシラジンの11.6 mg / kgを182 mg / kgでは0.9％NaClで1:2に希釈）を0.5mlと静脈麻酔する。 </li><li class="jove_content">つま先ピンチ反射を麻酔のレベルを確認するために使用される。必要に応じて追加の麻酔薬を投与する。 </li><li class="jove_content">ラットが麻酔の場合は、腹腔は、外科はさみで開かれます。ラット大動脈または大静脈から注射器に血液を10〜15ミリリットル描画することで放血される。 </li></ol>

<ol>
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ヴィンセントPoitoutは共同創立者であり、Bêtagenex社、商用サービスとして、この記事で紹介輸液モデルを提供し、契約研究機関から研究契約を受け取った。

先行研究の数が我々の知識に、げっ歯類ではグルコース（ 例えば 10-15）や脂質（ 例えば 16,17）の慢性的な注入を採用しているが、両方の燃料の組み合わせ注入はマウスだけ18で報告されている。ここで紹介する慢性注入モデルはラットの生物学的機能の様々な栄養過剰の影響を研究するためのいくつかの利点を提供しています。まず、遺伝的に肥満の齧...

慢性的に血液循環におけるグルコースおよび脂質レベルの上昇は、膵臓β細胞（口コミを含むが、これらに限定されないグルコース恒常性の維持に関与するいくつかの器官の機能を変更することによって、2型糖尿病の病因に寄与することが提案されている1）。慢性高血糖は、このように悪循環を作成し、2型糖尿病患者における血糖コントロールの悪化に寄与する、そもそも高​​血糖を生じたβ-細胞の欠損を悪化させること糖毒性の仮説断定。同様に、glucolipotoxicity仮説は、しばしば2型糖尿病で観察されたグルコースと脂質レベルの同時上昇は、β細胞に有害であることを提案している。 膵β細胞機能に慢性的に高い栄養素の有害な影響の細胞および分子メカニズムを解読するunderstandiの鍵である2型糖尿病の病因の1 ngの。そのために、多くの研究は、ランゲルハンスの単離された膵島またはクローン、インスリン分泌細胞株におけるインビトロにおける慢性栄養素過剰ex vivoでのメカニズムを検討した。培養細胞又は膵島において使用脂肪酸の濃度はほとんどインビボ 2 における β細胞の近傍の循環レベルと一致していないので、これらのモデル系で得られた知見の生物体全体への変換は、具体的には、複雑である。一方、栄養素過剰に応答して、β細胞障害機構は、糖尿病のZuckerラット脂肪3,4、スナネズミPsammomys obesus 5および高脂肪食によって例示さ、糖尿病のげっ歯類モデルにおいて検討されている給餌マウス6。これらのモデルは、しかし、本質的な代謝異常を特徴とし、血中グルコースの操作が容易に従順ではないおよび/またはより制御と少ない慢性設定で脂質レベル。そうしないと正常動物の日数期間における栄養レベルの循環変更できるようにするために、我々は生理的パラメータおよび機能に、私たちが単独で、または組み合わせて、脂質とグルコースの効果を調べることができ、正常ラットの慢性注入モデルを開発しました7,8。

<table border="1">
		<tbody>
		 <tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		 </tr>
		 <tr>
			<td>Saline 0.9%</td>
			<td>BD</td>
			<td>JB1324</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Dextrose 70%</td>
			<td>McKesson</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Intralipid 20%</td>
			<td>Fresenius Kabi</td>
			<td>JB6023</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Metricide (Glutaraldehyde 2.6%)</td>
			<td>Metrex</td>
			<td>11-1401</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Heparin Sodium 10,000 USP u/ml</td>
			<td>PPC</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Carprofen</td>
			<td>Metacam</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Glycopyrrolate</td>
			<td>Sandoz</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Isoflurane</td>
			<td>Abbott</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Chlohexidine 2%</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Alcohol 70%</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Iodine</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>PE-50</td>
			<td>BD</td>
			<td>427411</td>
			<td></td>
		 </tr>
		 <tr>
			<td>CO-EX T22</td>
			<td>Instech Solomon</td>
			<td>BCOEX-T22</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Connector 22G</td>
			<td>Instech Solomon</td>
			<td>SC22/15</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Swivel 22G</td>
			<td>Instech Solomon</td>
			<td>375/22PS</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Y-Connector 22G</td>
			<td>Instech Solomon</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Counterbalance and arm</td>
			<td>Instech Solomon</td>
			<td>CM375BP</td>
			<td></td>
		 </tr>
		 <tr>
			<td>23 G blunted needles</td>
			<td>Instech Solomon</td>
			<td>LS23</td>
			<td></td>
		 </tr>
		 <tr>
			<td>23 G canulation pins</td>
			<td>Instech Solomon</td>
			<td>SP23/12</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Tethers (12 inch)</td>
			<td>Lomir</td>
			<td>RT12D</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Infusion jackets </td>
			<td>Lomir</td>
			<td>RJ01, RJ02, RJ03, RJ04</td>
			<td></td>
		 </tr>
		 <tr>
			<td>(SM-XL)</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Tether attachment piece</td>
			<td>Lomir</td>
			<td>RS T1</td>
			<td></td>
		 </tr>
		 <tr>
			<td>60 ml syringe</td>
			<td>BD</td>
			<td>309653</td>
			<td></td>
		 </tr>
		 <tr>
			<td>1 ml syringe</td>
			<td>BD</td>
			<td>309602</td>
			<td></td>
		 </tr>
		</tbody>
 </table>

a model of chronic nutrient infusion in the rat

栄養素の過剰なレベルへの慢性暴露は、いくつかの臓器や組織の機能に影響を与えると2型糖尿病など、肥満やメタボリックシンドロームに関連する多くの合併症の発展に寄与すると仮定される。グルコースおよび脂肪酸の過剰レベルは膵臓β細胞およびインスリンの分泌に影響を及ぼす機序を研究するために、我々は、ラットにおける慢性的な栄養輸液モデルを確立した。 7日間の回復期間を可能にする。、スイベルと、ケージ内で自由に移動することを可能にする動物カウンタウェイトシステムを使用してポンプにカテーテルを接続すると、注入する手順は全身麻酔下右頸静脈、左頸動脈をcatheterizingから成りグルコース及び/又は72時間、イントラリピッド（ヘパリンを注入したときに約80％unsaturated/20％飽和脂肪酸の混合物を生成する大豆油エマルジョン）。このモデルは、いくつかのアドバンタを提供しています薬理学的化合物を共注入するためのオプションが、前記食物モデルとは対照的に、比較的短い時間枠細かくグルコースおよび脂肪酸の循環目標レベルを調節する可能性を含むGES。なお、臓器の様々な栄養素誘発性機能不全のメカニズムを検討し、この文脈において、薬物の有効性をテストするために使用することができる。

手術を受けた42匹のラットの系列のうち、5匹は術後期間中に失われ、1ラットは86％の全体的な成功率を表す、注入中に失われた。最終的に注入した37匹のラットの平均体重は608だった±5手術前にgおよび588±注入開始時の6グラム（平均±SEであり、n = 37、P &lt;0.0001を対応のあるt検定によって）。以下の代表的な結果は、2注入群で得られた。生理食塩水（SAL）、及びグルコース+イントラリピッド...

Watch this Scientific Journal Video about A Model of Chronic Nutrient Infusion in the Rat at JoVE.com

A Model of Chronic Nutrient Infusion in the Rat

ラットにおけるグルコースとイントラリピッドの慢性的な注入のためのプロトコルが記載されている。このモデルは、臓器機能および生理学的パラメータに対する過剰な栄養素の影響を研究するために使用することができる。

ラットにおける慢性栄養注入のモデル

Chronic exposure to excessive levels of nutrients is postulated to  affect the function of several organs and tissues and to contribute to the  ...

a-model-of-chronic-nutrient-infusion-in-the-rat

Research

JoVE Journal

Medicine

12.6K ビュー.  CRCHUM. ラットにおけるグルコースとイントラリピッドの慢性的な注入のためのプロトコルが記載されている。このモデルは、臓器機能および生理学的パラメータに対する過剰な栄養素の影響を研究するために使用することができる。

ビデオ: A Model of Chronic Nutrient Infusion in the Rat

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

同所性大動脈移植：慢性血管障害の発生研究のラットモデル

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

医学

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

マウスの膀胱痛に対するモデルとしての膀胱膨満誘発内臓運動応答

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

Slow-release Drug Delivery through Elvax 40W to the Rat Retina: Implications for the Treatment of Chronic Conditions

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these diseases. Chronic retinal diseases such as retinal oedema or neovascularization usually require multiple intraocular injections to effectively treat the condition. However, the risks associated with these injections increase with repeated delivery of the drug. Therefore, alternative delivery methods need to be established in order to minimize the risks of reinjection. Several other investigations have developed methods to deliver drugs over extended time, through materials capable of releasing chemicals slowly into the eye. In this investigation, we outline the use of Elvax 40W, a copolymer resin, to act as a vehicle for drug delivery to the adult rat retina. The resin is made and loaded with the drug. The drug-resin complex is then implanted into the vitreous cavity, where it will slowly release the drug over time. This method was tested using 2-amino-4-phosphonobutyrate (APB), a glutamate analogue that blocks the light response of the retina. It was demonstrated that the APB was slowly released from the resin, and was able to block the retinal response by 7 days after implantation. This indicates that slow-release drug delivery using this copolymer resin is effective for treating the retina, and could be used therapeutically with further testing.

This paper details how Elvax 40W can be used as a slow-release method for drug delivery to the adult rat retina. The protocol for preparing, loading, and delivering the drug-resin complex to the eye is described.

ラット網膜へのエルバックス40Wを通じて徐放性ドラッグデリバリー：慢性症状の治療のための含意

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these ...

Neurodegeneration in an Animal Model of Chronic Amyloid-beta Oligomer Infusion Is Counteracted by Antibody Treatment Infused with Osmotic Pumps

Decline in hippocampal-dependent explicit memory (memory for facts and events) is one of the earliest clinical symptom of Alzheimer&#39;s disease (AD). It is well established that synapse loss and ensuing neurodegeneration are the best predictors for memory impairments in AD. Latest studies have emphasized the neurotoxic role of soluble amyloid-beta oligomers (A&#946;o) that begin to accumulate in the human brain approximately 10 to 15 yr before the clinical symptoms become apparent. Many reports indicate that soluble A&#946;o correlate with memory deficits in AD models and humans. The A&#946;o-induced neurodegeneration observed in neuronal and brain slice cultures has been more challenging to reproduce in many animal models. The model of repeated A&#946;o infusions shown here overcome this issue and allow addressing two key domains for developing new disease modifying therapies: identify biological markers to diagnose early AD, and determine the molecular mechanisms underpinning A&#946;o-induced memory deficits at the onset of AD. Since soluble A&#946;o aggregate relatively fast into insoluble A&#946; fibrils that correlate poorly with the clinical state of patients, soluble A&#946;o are prepared freshly and injected once per day during six days to produce marked cell death in the hippocampus. We used cannula specially design for simultaneous infusions of A&#946;o and continuous infusion of A&#946;o antibody (6E10) in the hippocampus using osmotic pumps. This innovative in vivo method can now be used in preclinical studies to validate the efficiency of new AD therapies that might prevent the deposition and neurotoxicity of A&#946;o in pre-dementia patients.

To study the effects of A&#946;o in vivo, we developed a model based on repeated hippocampal infusions of soluble A&#946;o coupled with continuous infusion of A&#946;o antibody (6E10) in the hippocampus using osmotic pumps to counteract the neurotoxic effect of A&#946;o.

慢性アミロイドβオリゴマー輸液の動物モデルにおける神経変性は、浸透圧ポンプを用いて抗体治療フューズによって打ち消されます

Decline in hippocampal-dependent explicit memory (memory for facts and events) is one of the earliest clinical symptom of Alzheimer&#39;s disease (AD). It ...

Use of a Central Venous Line for Fluids, Drugs and Nutrient Administration in a Mouse Model of Critical Illness

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce sepsis with the use of a central venous line for fluids, drugs and nutrient administration to mimic the human clinical setting. Critically ill patients require intensive medical support in order to survive. While the majority of patients will recover within a few days, about a quarter of the patients need prolonged intensive care and are at high risk of dying from non-resolving multiple organ failure. Furthermore, the prolonged phase of critical illness is hallmarked by profound muscle weakness, and endocrine and metabolic changes, of which the pathogenesis is currently incompletely understood. The most widely used animal model in critical care research is the cecal ligation and puncture model to induce sepsis. This is a very reproducible model, with acute inflammatory and hemodynamic changes similar to human sepsis, which is designed to study the acute phase of critical illness. However, this model is hallmarked by a high lethality, which is different from the clinical human situation, and is not developed to study the prolonged phase of critical illness. Therefore, we adapted the technique by placing a central venous catheter in the jugular vein allowing us to administer clinically relevant supportive care, to better mimic the human clinical situation of critical illness. This mouse model requires an extensive surgical procedure and daily intensive care of the animals, but it results in a relevant model of the acute and prolonged phase of critical illness.

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce sepsis with the use of a central venous line for fluids, drugs and nutrient administration to mimic the human clinical setting.

重大な病気のマウスモデルにおける流体、医薬品や栄養管理のための中心静脈ラインの使用

This protocol describes a centrally catheterized mouse model of prolonged critical illness. We combine the cecal ligation and puncture method to induce ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

ワイヤー図法を使用してマウスの腸間膜動脈の等尺性収縮力測定

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A New Best Practice for Validating Tail Vein Injections in Rat with Near-infrared-Labeled Agents

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part of many experimental procedures where reagents need to be introduced directly into the bloodstream. Unwittingly, the injection can be subcutaneous, possibly altering the scientific outcomes. Utilizing a nanoemulsion-based biological probe with an incorporated near-infrared fluorescent (NIRF) dye, this method offers the capability of imaging a successful tail vein injection in vivo. With the use of a NIRF imager, images are taken before and after the injection of the agent. An acceptable IV injection is then qualitatively or quantitatively determined based on the intensity of the NIRF signal at the site of injection.

Here we present a method to validate tail vein injections in rats by utilizing near-infrared fluorescence imaging data from dyes incorporated into agents or biological probes. The tail is imaged before and after the injection, the fluorescent signal is quantified, and an assessment of the injection quality is made.

近く-赤外線-ラベル エージェントとラットの尾静脈注射を検証するための新しいベスト プラクティス

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part ...

Acupuncture in a Rat Model of Asthma

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes methods for the fabrication of the high platform, establishes a rat model of asthma and measures changes in respiratory function using a noninvasive and real-time whole-body plethysmography (WBP) system.

喘息ラットモデルにおける鍼治療

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes ...

A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. Several rodent models have been developed to study CP, of which the bile duct 2,4,6 -trinitrobenzene sulfonic acid (TNBS) infusion model replicates the features of neuropathic pain seen in CP. However, bile duct drug infusion in mice is technically challenging. This protocol demonstrates the procedure of bile duct TNBS infusion for generation of a CP mouse model. TNBS was infused into the pancreas through the ampulla of Vater in the duodenum. This protocol optimized drug volume, surgical techniques, and drug handling during the procedure. TNBS-treated mice showed features of CP as reflected by bodyweight and pancreas weight reductions, changes in pain-associated behaviors, and abnormal pancreatic morphology. With these improvements, mortality associated with TNBS injection was minimal. This procedure is not only critical in generating pancreatic disease models but is also useful in local pancreatic drug delivery.

Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

胆管TNBS輸液による慢性膵炎のマウスモデル

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. ...

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

温虚血/再灌流損傷のラット肺移植モデル:転帰を改善するための最適化

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...