著者は、技術支援のためにクリスティPahrmannに感謝します。 SSはドイツ学術振興（DFG）（SCHR992/3-2とSCHR992/4-2）から資金提供を受けた。

全ての動物研究は、関連する動物ケアガイドラインに従って行われるべきである。前初めプロトコルへの動物の仕事のための制度的認可を受けなければならない。 1。筋内膜増殖症モデル（A） 内膜肥厚モデルの場合、約25グラムの重さ8週齢のC57BL/6J（在庫番号000664、C57BL/6J）マウスを購入。従来の条件の下で、これらの実験のための家のマウス;マウス標準マウス飼料を飼料と水を自由に提供する。 <ol><li> マウスの調製：イソフルラン（2％）が導入室を利用して麻酔状態にマウスを導入する。 <ol><li>ヘアトリマーを使用して腹部の毛を削除すると、その背中の上でマウスを置き、麻酔を保証するためにフェイスマスクで鼻と口をカバーしています。 </li><li>プロボ - ヨウ素と、普遍的に腹部を消毒、2サイクルで、80％エタノールが続く。 </li><li>存在しないことを保証する麻酔の十分な深さを監視するために後肢をつまんで反射の。 </li><li>腹部臓器を公開するには、2つのステップで白線に沿って皮膚と筋肉を分離する。 </li><li>生理食塩水しっとり手袋で腸を置く。湿ったそれらを保つために、腸の周りに手袋を包む。 </li><li>注意深く腹部大動脈天竺脂肪組織を離れてオフにします。任意の分岐血管を傷つけないように注意しながら、ダウンの分岐部に大動脈大動脈を公開します。 </li><li>まず、血液の流れをシャットダウンするには、近位に大動脈大動脈を顕微クランプを置く。今、大動脈分岐の隣に第2の遠位クランプを適用します。 </li><li>血液の流れが中断されたときに、近位のクランプに近い大動脈に小さな切開を行います。 </li><li> 0.9％生理食塩水で湿らされたバルーンが先端に付いたカテーテルを挿入し、大動脈の遠位クランプに向かって前進する。 </li><li>内皮損傷の場合は、ballooを膨張させることによって大動脈をはぎ取るN慎重にゆっくりと近位クランプの方向にカテーテルを引っ張って。 </li><li>二回削剥演習を繰り返します。 </li><li>カテーテルを取り外し、10-0プロレンランニング縫合糸を使用して大動脈切開を閉じます。 </li><li>慎重に遠位クランプを開きます。大動脈切開部位での出血の場合には、必要に応じて、再度、クランプを閉じ、出血を見つけて、追加の中断ステッチを配置。 </li><li>出血が認められない場合は、ゆっくりと近位のクランプを開きます。 </li><li>大動脈パルスが切開部から遠位側に見えるはずです。今腸その場で戻って配置することができる。 </li><li>予め温め滅菌生理食塩水を用いた腹腔を洗浄します。 </li><li> 6-0プロレンランニング縫合糸を使用して、筋層で始まる腹壁を閉じます。 </li><li>次の5-0プロレン実行、実行中の縫合糸の使用をして肌を適応させる。 </li><li>マウスがまだ麻酔している間、皮下麟蹄を経由して4〜5 mg / kgのカルプロフェンを適用ction。 </li><li>疼痛コントロールのための飲料水（100ミリリットル当たり50ミリグラムメタミゾール）にメタミゾールを追加し、3日後に手術のために続けている。回復を通して動物を監視します。 注：このモデルの観察期間は28日間です。 </li></ol></li></ol> 2。アテローム性動脈硬化症モデル（B） 4週齢でマウス（在庫番号002052、B6.129P2-Apoetm1Unc / J）をした後、4〜6ヶ月間与え、アテローム性動脈硬化症のモデルでは、（ - / -アポ）アポリポタンパク質E欠損を購入。従来の条件の下で、これらの実験のための家のマウス;到着（ハーランラボラトリーズTD.88137）上にマウス高脂肪西洋食を供給し、4から6ヶ月の間、 水を自由に提供する。 <ol><li> マウスの調製： <ol><li>アポフィード - / - 西洋食付きマウスは生後4週から始まる。食餌操作は、到着時に直接開始する必要があります。 </li><li>実験期間を通して西洋食を維持している。&lt;/李&gt; </ol></li></ol> 3。分析<ol><li> 筋内膜増殖症モデル（A） このモデルにおける関心領域」は、病変領域」である。分析のためのオプションは、管腔閉塞を含み、I / M比（内膜/中膜比）は、最大myointima厚さ、筋内膜面積、線維症トリクローム染色、および単核細胞のためのH＆E染色（ 図1A〜1Cに示されるように）。科学的な質問によると、コラーゲン、免疫組織化学、および共焦点のメソッドのピクロシリウスレッドのような他の多くの染色を適用することができる。 </li><li> アテローム性動脈硬化症モデル（B） このモデルでの関心領域は、大動脈弁、大動脈弓だけでなく、下行大動脈である。分析のためのオプション：スーダンレッド脂質の染色や大動脈弁のトリクローム染色、大動脈弓と下行大動脈（ 図1（d）に示すように）。 このモデルはまた、他のような染色のさまざまなオプションをもたらす中性脂質については、例えばオイルレッドO、ピクロシリウスレッド、免疫組織化学、および共焦点方法。 <ol><li>スーダンレッド染色：新たに採取大動脈組織にスーダンレッド染色を行います。 <ol><li>収穫前にダイエット6Hを削除します。 </li><li>イソフルラン（2％）が導入室を利用して麻酔状態にマウスを導入する。麻酔の十分な深さを監視するために後肢をつまんで反射神経が存在しないことを保証する。 </li><li>胸を開きます。心尖​​を切断し、大動脈を修正するために大動脈基部を通じて4％PFA（3ミリリットル）に続いて、最初に生理食塩水（3ml）で、左心室を経由して灌流する。 </li><li>その分岐ルートから大動脈を解放します。可能な限りの脂肪組織を除去するようにしてください。 </li><li>心臓や大動脈根を切断し、組織学のために4％PFAに格納します。 </li><li>組織学のために大動脈基部は、AORの切断を可能にするために慎重に直立位置にパラフィンブロック内に配置されなければならないすべての3弁膜チラシを表示TICルート生成スライド。 </li><li>大動脈ツリーの一方の端をカットし、細かいピンとシリコーン層の上に固定します。縦に大動脈を開き、大動脈組織を修正。 </li><li> 50％エタノールを用いて組織を洗浄します。約15分間、スダンIII染色溶液で組織を浸す。 </li><li>過剰スーダンIIIを除去するために50％エタノールで軽くすすいでください。 （注：50％エタノールへは、染料を洗うこと）をdH 2 Oですすぎ、そしてさらなる分析のために写真を撮る。 </li></ol></li></ol></li></ol>

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myointimal+hyperplasia:+pathogenesis+and+implications.+2.+Animal+injury+models+and+mechanical+factors.">Myointimal hyperplasia: pathogenesis and implications. 2. Animal injury models and mechanical factors.</a> Artificial organs. 15, 103-118 (1991).</li><li>Nitschke, Y., Weissen-Plenz, G., Terkeltaub, R., Rutsch, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Npp1+promotes+atherosclerosis+in+ApoE+knockout+mice.">Npp1 promotes atherosclerosis in ApoE knockout mice.</a> Journal of cellular and molecular. 15, 2273-2283 (2011).</li></ol>

著者らは、開示することは何もありません。

両疾患の患者で同様の症状を引き起こすが、プラーク発生の根本的なメカニズム、したがって、治療アプローチは、2非常に異なっている。動脈狭窄の様々な形態での患者の臨床所見だけでなく、プラークの開発期間は、基本的なメカニズムに依存しています。 患者の臨床所見に応じて、サンプル分析の異なる方法が5,15を実行する必要があります。これ?...

先進国における虚血性心疾患は死亡率1の主要な原因のまま。冠動脈狭窄の原因は多種多様であり、最も一般的な治療戦略2を 、残りの血行再建術と筋内膜増殖症だけでなく、アテローム性動脈硬化症が含まれる。明らかに内膜肥厚やアテローム性動脈硬化症の機構経路を区別する研究モデルは、動脈プラークの開発における病理生物学的および病態生理学的プロセスを調査するために不可欠である。このビデオでは筋内膜増殖症やアテローム性動脈硬化症の発症のどちらかを研究するために使用される二つの異なるマウスモデルを導入する。 筋内膜増殖症、アテローム性動脈硬化症は、もともと3について記載の「レスポンス·ツー·傷害」パラダイムを経由して形成することが想定される。内皮層の機械的破壊は、パラクリン効果によって増強激しいリモデリングをもたらす。いくつかのディがありますfferent動物モデルは、一般的に筋内膜過形成を研究するために使用。いくつかのグループは、ラット4の上行大動脈に金属のデバイスを使用しています。バルーンカテーテルを用いて行わ大動脈削剥モデルは、一般に実験用ラット5,6で使用される。実験用マウス7で行われる内膜過形成モデルは、頸動脈の結紮を実装し、筋内膜病変8を誘導する。私たちの研究室では、腹部大動脈侵食モデル-筋内膜過形成の発達を研究する-だけでなく、ヒトのステント再狭窄モデル9は、一般的に使用されている。このビデオでは、適切な実験動物モデルを選択すると、動脈狭窄の機序や病態生理学的研究のために重要であることを強調している。 筋内膜過形成モデルは、アテローム性動脈硬化症のモデルと区別されなければならない。後者については、（アポE - / - ）アポリポタンパク質E欠損西洋食と組み合わせたマウスは一般にatherosclを誘導するために使用されているHな病変10,11,12,13。マウスでの筋内膜疾患のこれらのタイプの両方を誘導するための例としては、血管狭窄14を分析するために別のモデルに適応したオプションと一緒に、ここに示されています。

<table border="0" cellpadding="0" cellspacing="0" style="width:617px;" width="618">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="17">
			<td height="17" style="height:17px;width:156px;">Name</td>
			<td style="width:183px;">Company</td>
			<td style="width:139px;">Catalog number</td>
			<td style="width:140px;">Comments</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Thyoglycollate Broth 3%</td>
			<td>Fluka</td>
			<td align="right">70157</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">PFA 4%</td>
			<td>Electron Microscopy Sciences</td>
			<td>#157135S</td>
			<td align="right">20%</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sudan III staining solution</td>
			<td>Sigma Aldrich</td>
			<td>S4131</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse C57BL/6J</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock # 0006664</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse ApoE-/-</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock #002052</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Western Diet</td>
			<td>Harlan Laboratories</td>
			<td>TD.88137</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">hair clipper</td>
			<td>WAHL</td>
			<td>8786-451A ARCO SE</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Forene</td>
			<td>Abbott</td>
			<td></td>
			<td>Isoflurane</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">microsurgical clamp</td>
			<td>Fine Science Tools</td>
			<td>18055-04</td>
			<td>Micro-Serrefine - 4mm</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">clamp applicator</td>
			<td>Fine Science Tools</td>
			<td>18056-14</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">catheter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">10-0 prolene suture</td>
			<td>Ethicon</td>
			<td>788G</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">6-0 prolene suture</td>
			<td>Ethicon</td>
			<td>8709H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">5-0 prolene suture</td>
			<td>Ethicon</td>
			<td>EH7229H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Rimadyl</td>
			<td>Pfizer&#160;</td>
			<td></td>
			<td>Carprofen</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Metacam 1.5mg/ml</td>
			<td>Boehringer Ingelheim</td>
			<td></td>
			<td>Metamizol</td>
		</tr>
	</tbody>
</table>

inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models

虚血性心疾患、例えば観察される動脈狭窄を誘導するための種々 のin vivo実験用げっ歯類モデルは、動脈プラーク形成および狭窄を含む疾患を模倣するように確立されている。 2つの高度に再現可能なマウスモデル - 両方の動脈狭窄をもたらしたが、それぞれが開発のさまざまな経路を根底には - ここで紹介されています。モデルは、動脈狭窄の2の最も一般的な原因を表し;すなわち、1マウス各筋内膜過形成のためのモデル、およびアテローム性動脈硬化症が示されている。筋内膜過形成を誘発するために、腹部大動脈のバルーンカテーテル傷害が行われる。動脈硬化性プラークの発展のために、アポE - / - 西脂肪の食事と組み合わせたマウスモデルが使用されます。結果の測定·評価のための別のモデルに適合したオプションについては、この原稿に名前が付けられ、説明されています。これらの二つのモデルを導入することと比較すると、のinformatを提供しています科学者たちは尋ねた科学的な質問への応じた適切な動脈狭窄モデルを選択するためのイオン。

図1は、両方のモデルについて異なる分析オプションおよび誘導疾患状態を示している。内膜過形成モデルでは、トリクローム染色の代表断面の分析が示されている。これらの断面から、I / M比は、最大プラーク厚さが、内腔の最大内膜厚さ、パーセント管腔閉塞、ならびにプラーク面積のような結果を測定し、計算することができる。 アテローム性動脈硬?...

Watch this Scientific Journal Video about Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models at JoVE.com

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

このビデオでは、マウスの動脈の内膜のプラークの開発の2つのモデルを示し、筋内膜増殖症およびアテローム性動脈硬化症の違いを強調している。

マウスにおけるアテローム性動脈硬化症に対する筋内膜過形成を誘発する。つの有効なモデルの導入

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque ...

inducing-myointimal-hyperplasia-versus-atherosclerosis-mice-an

Research

JoVE Journal

Medicine

12.0K ビュー.  University Hospital Hamburg. このビデオでは、マウスの動脈の内膜のプラークの開発の2つのモデルを示し、筋内膜増殖症およびアテローム性動脈硬化症の違いを強調している。 

ビデオ: Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

マウスでのせん断応力誘発アテローム性動脈硬化症をトリガするための頸動脈カフの注入

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

医学

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

マウス大動脈洞におけるアテローム性動脈硬化症の病変の定量分析とキャラクタリゼーション

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Acute Brain Trauma in Mice Followed By Longitudinal Two-photon Imaging

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no effective treatment for it yet. Limitations of currently used animal models impede understanding of the pathology mechanism. Multiphoton microscopy allows studying cells and tissues within intact animal brains longitudinally under physiological and pathological conditions. Here, we describe two models of acute brain injury studied by means of two-photon imaging of brain cell behavior under posttraumatic conditions. A selected brain region is injured with a sharp needle to produce a trauma of a controlled width and depth in the brain parenchyma. Our method uses stereotaxic prick with a syringe needle, which can be combined with simultaneous drug application. We propose that this method can be used as an advanced tool to study cellular mechanisms of pathophysiological consequences of acute trauma in mammalian brain in vivo. In this video, we combine acute brain injury with two preparations: cranial window and skull thinning. We also discuss advantages and limitations of both preparations for multisession imaging of brain regeneration after trauma.

Acute brain trauma is a severe injury that has no adequate treatment to date. Multiphoton microscopy allows studying longitudinally the process of acute brain trauma development and probing therapeutical strategies in rodents. Two models of acute brain trauma studied with in vivo two-photon imaging of brain are demonstrated in this protocol.

縦二光子イメージングに続いてマウスの急性脳外傷

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

マウスにおける心筋梗塞を誘発する低侵襲手術手順

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Surgical Models of Gastroesophageal Reflux with Mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to induce reflux of gastric contents, duodenal contents or mixed contents. Surgical procedures and general principles have been described in detail. A researcher with surgical experience should be able to grasp the technique after a short period of practice. After surgery, most mice can survive and develop reflux esophagitis similar to that in humans. However, it should be noted that histological differences between mouse and human esophagus are the inherent limitations of these surgical models. If used for research on Barrett&#8217;s esophagus and adenocarcinoma, these procedures may need to be combined with genetic modifications.

This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett&#8217;s esophagus and esophageal adenocarcinoma.

マウスを用いた胃食道逆流の外科モデル

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

マウスの気管の迅速な挿管のための改善された方法

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the brain for visual perception. Damage to RGCs and their axons leads to vision loss and/or blindness. Although the specific cause of glaucoma is unknown, the primary risk factor for the disease is an elevated intraocular pressure. Glaucoma-inducing procedures in animal models are a valuable tool to researchers studying the mechanism of RGC death. Such information can lead to the development of effective neuroprotective treatments that could aid in the prevention of vision loss. The protocol in this paper describes a method of inducing glaucoma - like conditions in an in vivo rat model where 50 &#181;l of 2 M hypertonic saline is injected into the episcleral venous plexus. Blanching of the vessels indicates successful injection. This procedure causes loss of RGCs to simulate glaucoma. One month following injection, animals are sacrificed and eyes are removed. Next, the cornea, lens, and vitreous are removed to make an eyecup. The retina is then peeled from the back of the eye and pinned onto sylgard dishes using cactus needles. At this point, neurons in the retina can be stained for analysis. Results from this lab show that approximately 25% of RGCs are lost within one month of the procedure when compared to internal controls. This procedure allows for quantitative analysis of retinal ganglion cell death in an in vivo rat glaucoma model.

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is characterized by damage to retinal ganglion cells. Inducing glaucoma in animal models can provide insight into the study of this disease. Here, we outline a procedure that induces loss of RGCs in an in vivo rat model and demonstrates the preparation of whole-mount retinas for analysis.

緑内障誘導の手順<em&gt;インビボ</em&gt;ラットモデルと全体のマウント網膜準備

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

マウスにおける加速アテローム性動脈硬化症の誘導:ワイヤー傷害モデル

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

マウスの腹部大動脈 Myointimal 過形成を誘導するために気球を利用した傷害

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. Various approaches have been described, such as the inhalation of aerosolized LPS as well as nasal or intratracheal instillation. The presented protocol describes a detailed step-by-step procedure to induce ALI in mice by direct intratracheal LPS instillation and perform FACS analysis of blood samples, bronchoalveolar lavage (BAL) fluid, and lung tissue. After intraperitoneal sedation, the trachea is exposed and LPS is administered via a 22 G venous catheter. A robust and reproducible inflammatory reaction with leukocyte invasion, upregulation of proinflammatory cytokines, and disruption of the alveolo-capillary barrier is induced within hours to days, depending on the LPS dosage used. Collection of blood samples, BAL fluid, and lung harvesting, as well as the processing for FACS analysis, are described in detail in the protocol. Although the use of the sterile LPS is not suitable to study pharmacologic interventions in infectious diseases, the described approach offers minimal invasiveness, simple handling, and good reproducibility to answer mechanistic immunological questions. Furthermore, dose titration as well as the use of alternative LPS preparations or mouse strains allow modulation of the clinical effects, which can exhibit different degrees of ALI severity or early vs. late onset of disease symptoms.

Presented here is a step-by-step procedure to induce acute lung injury in mice by direct intratracheal lipopolysaccharide instillation and to perform FACS analysis of blood samples, bronchoalveolar lavage fluid, and lung tissue. Minimal invasiveness, simple handling, good reproducibility, and titration of disease severity are advantages of this approach.

直接気管内リポ多糖刺激によるマウスの急性肺損傷の誘導

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...