We thank Drs. Xiaofang Wang and Katharina Hopp for their invaluable help with the animal models. This work has been supported by grants from the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health (DK090728, DK058816).

生きた動物との任意の手順を開始する前に、実験プロトコルは、制度の動物の管理と使用委員会（IACUC）によって承認されなければなりません。 1.スキャナ設定<ol><li>開始する前に、ヒーターをOFFの位置にあることを確認してください。 </li><li>ミニイメージング勾配と38ミリメートルのRFコイルとミニイメージングホルダーを選択します。 </li><li>ホルダーの中央の穴に温度可変アセンブリをインストールします。 </li></ol> 2.動物の準備<ol><li> MRI実験のために、気化イソフルランを使用して、最適な麻酔を達成。麻酔の誘導のために、吸収性の組織が並んで誘導チャンバ内に動物を配置します。 2.0〜2.5 L / minとし、酸素中3％イソフルランにイソフルラン気化器の流量計を調整します。 </li><li>この段階では金属製のタグまたは他の金属のオブジェクトを削除します。麻酔下ながら乾燥を防止するために、動物の目に獣医軟膏を適用します。 </li><li>動物一度麻酔の外科的平面に到達した（ すなわち、つま先のピンチに引っ込め反射の損失）は、ノーズコーンに挿入し、その鼻とホルダに動物を配置します。酸素中の1.5から2.0パーセントに2.0〜2.5ミリリットル/分であり、イソフルラン濃度にプローブに麻酔の空気の流れを設定します。麻酔はプロシージャの間、ノーズコーンを介して配信されます。定期的に〜40 BPMの呼吸速度を維持するために、動物の年齢および体重に応じて、イソフルラン濃度を調整します。 </li><li>代わりに動物を確保し、MRI実験中の動きを防止するために、動物の所有者を使用します。スキャンするボディ領域に応じて、動物ホルダーの種類を変え。 注：実験室のプラスチック（ポリプロピレン、テフロン、ポリスチレン、ポリカーボネート）からカスタマイズされたホルダー（新生マウス〜160グラムのラットに）特定の実験に対応して、動物のサイズに合わせて作ることができます。 </li><li>動物の体温を監視するために、動物に直腸温度計を配置します。 EXPの間erimentは、暖かい空気の流れを使用して、35〜37℃で動物を維持します。動物の体の温度フィードバックに基づいて、空気の温度（30〜38℃）で、フロー（1,200-2,000 L /時間）を調整します。 </li><li>呼吸速度を監視するために、動物の腹部にバルーン呼吸圧力センサを取り付けます。 </li><li> RFコイルの中心に動物を固定し、慎重にMRIスキャナに動物とRFコイルを配置します。 </li></ol> 3. MRI実験<ol><li>使用されるRF電力を最小化し、信号対雑音比を最大にするために、実験を開始する前に、RFコイルとチューニングと一致します。マッチング/チューニングを開始するには： <ol><li>ツールアイコンをクリックして、分光器制御ツールを開きます。 </li><li>分光器制御ツールで取得→揺れをクリックします。 ACQ /レコウィンドウがウォブル曲線を表示する開きます。 </li><li>あるいは、反射されたRFパワーまで小さなステップでチューニングとマッチングコンデンサ（チューニングを使用し、ロッドにマッチする）を調整最小化されます。目標は、横軸にゼロに位置する垂直軸で最小の曲線を確認することです。 </li><li>コイルのキャリブレーションが正常に達成されたときは、ACQ /レコウィンドウで[停止]ボタンを押してください。 </li></ol></li><li> 、軸方向コロナル及びサジタル画像を作成するために、3つの直交面にスカウト画像を取得します。スカウト画像12を取得するために、このようなイントラゲートファーストローアングルショット（IG-FLASH）として、高速画像シーケンスを使用してください。実際の撮影のために適切なジオメトリを設定するためにスカウト画像を使用してください。 </li><li>特定の研究目的に応じて、適切な画像シーケンスとパラメータを選択し、トラフィックの光でスキャンを開始します。これは、RFチャネルを校正磁石をシム、水のために共鳴キャリア周波数を設定し、すべて自動的に受信機の利得を調整します。 <ol><li>解剖学的研究およびT2強調画像では、2Dマルチスライスまたは3Dモードで取得します。所与の空間解像度のための実験時間を短縮するために、フィールドを維持します-ofビュー（FOV）可能ですが、ラップアラウンドの成果物（2.56〜3.2センチメートル）を回避するのに十分な大きさと小さいです。 </li></ol></li><li>繰り返し時間（TR）、および/またはスライスの数を適切に選択することによって、動物の呼吸周期より若干短い選択された配列の周期にしてください。これは、データが動物の沈黙期間中に収集されることを保証します。 <ol><li>例えば、腹部の画像について、〜30 BPMでの動物の呼吸速度を維持します。それは息当たり約2,000ミリ秒です。 TRで、リラクゼーション強化（RARE）シーケンスでターボ迅速な取得を使用して、11-19冠状スライスを取得/ TE 9分の1500ミリ秒、RARE因子8と（マトリクス256×256、FOV 2.56 X 2.56センチメートル、スライス厚0.75ミリメートル） 。 注：1,500ミリ秒にTRを調整し、〜30 BPM（ブレス2,000ミリ秒）動物の呼吸速度を維持することによって、我々はデータが動物の沈黙期間中に収集されていることを確認してください。 </li></ol></li><li>すべての画像の取得が完了した後、スキャンされた動物を置き加熱したパッドの上に歩行するまで監視します。回復後、ケージに動物を返し、動物施設に戻す前に1時間以上で監視。 </li></ol>

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The authors have nothing to disclose.

この原稿は、PKDのための齧歯類モデルにおける in vivo表現型の特徴付けや薬物モニタリングのためのツールとしてUHF MRIを使用することの実現可能性を示しています。 我々は、マイクロ、ミニイメージングアクセサリーを装備した大口径アヴァンセIII高分解能NMR分光計で16.4 Tで行った実験について説明します。分光計はParavision 5.1イメージングソフトウェアに...

多発性嚢胞腎（PKD）は、腎嚢胞の発生を特徴単一遺伝子疾患のグループが含まれています。これらの中でも最も一般的なタイプ1,2を表す常染色体優性多発性嚢胞腎疾患（ADPKD）および常染色体劣性多発性嚢胞腎（ARPKD）が、あります。 ADPKD、遺伝性腎嚢胞性疾患の最も頻繁な形態は、PKD1またはPKD2遺伝子の変異によって発信されます。これは、遅発性、複数の両側腎嚢胞、可変超腎嚢胞を伴うだけでなく、心血管および筋骨格異常によって特徴付けられます。 ARPKDは、最も一般的に新生児や幼児に影響を与え、PKHD1の突然変異によって引き起こされ、拡大されたエコー源性腎臓および先天性肝線維症3によって特徴づけられます。 重要なことは、ADPKDは、両方の実質的なPをもたらす遺伝子（ジェニック）および変異（対立遺伝子）レベルで、不均一性によって特徴付けられますhenotypic変動。 PKD1遺伝子の変異は重篤な臨床プレゼンテーション（多数の嚢胞、早期診断、高血圧、および血尿）、ならびに（PKD2変異を有する患者よりも20年前に）腎疾患段階を終了する急速な進行と関連している4。重度の多嚢胞肝疾患（PLD）と血管異常PKD1とPKD2 5の両方で突然変異と関連することができます。 ADPKDの腎臓合併症の大部分は、関連する炎症および線維症に伴う嚢胞の拡大の結果として主に生じます。嚢胞の開発は子宮内で開始し、患者の生涯を通じて継続します。腎臓は通常、彼らが20回以上正常な腎臓のボリュームに達する可能性があるにもかかわらず、自分の腎臓形の形状を維持します。腎嚢胞の患者本二国間分布のほとんどが、いくつかのまれなケースでは、嚢胞は、一方的または非対称パターンで発症することがあります。 主要なチャレンジVol。腎臓専門医がADPKDの患者を次または療法を実施するためのGEは、疾患の自然史です。そのコースの大部分の間、腎機能が正常なままであり、腎機能が低下し始める時点で、腎臓のほとんどは、嚢胞に置き換えられています。治療は、後の段階で実装されている場合には、患者は既に慢性腎疾患におけるノーリターンのポイントに達している可能性があるので成功しにくいです。治療を早期に開始されたときにこれとは対照的に、それは単に、糸球体濾過率に基づいて応答を識別することは困難です。その結果、疾患進行のマーカーとして腎体積の概念が注目を集め。 多発性嚢胞腎疾患（CRISP）研究の放射線学的イメージング研究コンソーシアムは、ADPKD患者で腎臓や嚢胞体積の増加が直接的に総腎容積（TKV）の可能性を強調し、腎機能の低下と相関することが示されています病気の進行のために6,7 urrogateマーカー。したがって、TKV現在ADPKD 2,8,9のための複数の臨床試験において、一次または二次エンドポイントとして使用されます。 複数のマウスモデルなどの自然突然変異と遺伝的に操作がPKD 10,11の病因に光を当てるしています。 PKD1またはPKD2モデル（PKD1またはPKD2のいずれかにおける突然変異）は、それらの完全に模倣ヒト疾患として、最も人気のあるものとなっています。また、PKD1またはPKD2の遺伝子以外の遺伝子に突然変異を有するげっ歯類モデルは、疾患に関連するシグナル伝達経路を解明するための実験プラットフォームとして使用されてきました。さらに、これらのモデルのいくつかは、潜在的な治療法を試験するために使用されてきました。しかし、PKDのための多くのげっ歯類の研究における制限因子は、多くの場合、順次腎臓における解剖学的および機能的変化を分析するための効率的な非侵襲的な方法がないことです。 磁気Resonanceイメージング（MRI）は、ADPKD患者に追従するために、現在の金標準的な画像化技術である、優れた軟部組織コントラストと解剖学的詳細を提供し、TKVの測定を可能にします。 MRIがよく、インビボでより大きな動物およびヒト、イメージング小げっ歯類において解剖学的撮像のために確立されても高解像度の画像を取得する能力は、その有用性を制限することができる追加の技術的課題を伴います。超高磁場の導入（UHF）MRI（7から16.4 T）と強い勾配の開発と、それはそれと同じような診断品質と高い信号対雑音比およびMRI画像の空間分解能を達成することが可能になりましたヒトにおいて得られました。したがって、PKDのための小さな齧歯類モデルの in vivoイメージングのためのUHF MRIの使用は、研究者のための強力なツールとなっています。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>AVANCEIII-700 (16.4 T)</td>
			<td>Bruker</td>
			<td>BH067206</td>
			<td>Wide-bore two channel multinuclear spectrometer equipped with mini and micro-imaging accessories for in vivo small rodent imaging</td>
		</tr>
		<tr>
			<td>TopSpin2.0PV&#160;</td>
			<td>Bruker</td>
			<td>H9088TA2</td>
			<td>Spectrometer processing software&#160;</td>
		</tr>
		<tr>
			<td>Paravision 5.1&#160;</td>
			<td>Bruker</td>
			<td>T10314L5</td>
			<td>Imaging sofware</td>
		</tr>
		<tr>
			<td>VTU BVT 3000 digital</td>
			<td>Bruker</td>
			<td>W1101095</td>
			<td>Temperature controller</td>
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use of ultra-high field mri in small rodent models of polycystic kidney disease for in vivo phenotyping and drug monitoring

Several in vivo pre-clinical studies in Polycystic Kidney Disease (PKD) utilize orthologous rodent models to identify and study the genetic and molecular mechanisms responsible for the disease, and are very convenient for rapid drug screening and testing of promising therapies. A limiting factor in these studies is often the lack of efficient non-invasive methods for sequentially analyzing the anatomical and functional changes in the kidney. Magnetic resonance imaging (MRI) is the current gold standard imaging technique to follow autosomal dominant polycystic kidney disease (ADPKD) patients, providing excellent soft tissue contrast and anatomic detail and allowing Total Kidney Volume (TKV) measurements.A major advantage of MRI in rodent models of PKD is the possibility for in vivo imaging allowing for longitudinal studies that use the same animal and therefore reducing the total number of animals required. In this manuscript, we will focus on using Ultra-high field (UHF) MRI to non-invasively acquire in vivo images of rodent models for PKD. The main goal of this work is to introduce the use of MRI as a tool for in vivo phenotypical characterization and drug monitoring in rodent models for PKD.

本稿では、我々はPKDおよびその他の腎疾患のげっ歯類モデルにおいてin vivoでの表現型の特徴付けや薬物モニタリングのためのツールとしてUHF MRIの有用性を示すことを目指しています。全ての実験は、IACUCにより承認実験プロトコルの一部でした。 UHF MRIを使用してPKDのための小さな齧歯類モデルの in vivoでの表現型の場合 ： ...

Watch this Scientific Journal Video about Use of Ultra-high Field MRI in Small Rodent Models of Polycystic Kidney Disease for In Vivo Phenotyping and Drug Monitoring at JoVE.com

Use of Ultra-high Field MRI in Small Rodent Models of Polycystic Kidney Disease for In Vivo Phenotyping and Drug Monitoring

The use of ultra-high field MRI as a non-invasive way to obtain phenotypic information of rodent models for polycystic kidney disease and to monitor interventions is described. Compared with the traditional histological approach, MRI images can be acquired in vivo, allowing for longitudinal follow-up.

のための多発性嚢胞腎疾患の小齧歯類モデルにおける超高磁場MRIの使用<em&gt;インビボ</em&gt;表現型と薬物モニタリング

Several in vivo pre-clinical studies in Polycystic Kidney Disease (PKD) utilize orthologous rodent models to identify and study the genetic and molecular ...

use-ultra-high-field-mri-small-rodent-models-polycystic-kidney

Research

JoVE Journal

Medicine

Use of Ultra-high Field MRI in Small Rodent Models of Polycystic Kidney Disease for In Vivo Phenotyping and Drug Monitoring

11.4K ビュー.  Mayo Clinic. The use of ultra-high field MRI as a non-invasive way to obtain phenotypic information of rodent models for polycystic kidney disease and to monitor interventions is described. Compared with the traditional histological approach, MRI images can be acquired in vivo, allowing for longitudinal follow-up.

ビデオ: Use of Ultra-high Field MRI in Small Rodent Models of Polycystic Kidney Disease for In Vivo Phenotyping and Drug Monitoring

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

Muscle strains are one of the most common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and physical exam alone, however the clinical presentation can vary greatly depending on the extent of injury, the patient's pain tolerance, etc. In patients with muscle injury or muscle disease, assessment of muscle damage is typically limited to clinical signs, such as tenderness, strength, range of motion, and more recently, imaging studies. Biological markers, such as serum creatine kinase levels, are typically elevated with muscle injury, but their levels do not always correlate with the loss of force production. This is even true of histological findings from animals, which provide a "direct measure" of damage, but do not account for all the loss of function. Some have argued that the most comprehensive measure of the overall health of the muscle in contractile force. Because muscle injury is a random event that occurs under a variety of biomechanical conditions, it is difficult to study. Here, we describe an in vivo animal model to measure torque and to produce a reliable muscle injury. We also describe our model for measurement of force from an isolated muscle in situ. Furthermore, we describe our small animal MRI procedure.

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

An in vivo animal model of injury is described. The method takes advantage of the subcutaneous position of the fibular nerve. Velocity, timing of muscle activation, and arc of motion are all pre-determined and synchronized using commercial software. Post injury changes are monitored in vivo using MR imaging/spectroscopy.

アン in vivoで</em収縮で誘起された損傷と回復の非侵襲的モニタリングの>齧歯類モデル

Muscle strains are one of the most  common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and  physical ...

医学

Spectral Karyotyping to Study Chromosome Abnormalities in Humans and Mice with Polycystic Kidney Disease

Conventional method to identify and classify individual chromosomes depends on the unique banding pattern of each chromosome in a specific species being analyzed 1, 2. This classical banding technique, however, is not reliable in identifying complex chromosomal aberrations such as those associated with cancer. To overcome the limitations of the banding technique, Spectral Karyotyping (SKY) is introduced to provide much reliable information on chromosome abnormalities.
SKY is a multicolor fluorescence in-situ hybridization (FISH) technique to detect metaphase chromosomes with spectral microscope 3, 4. SKY has been proven to be a valuable tool for the cytogenetic analysis of a broad range of chromosome abnormalities associated with a large number of genetic diseases and malignancies 5, 6. SKY involves the use of multicolor fluorescently-labelled DNA probes prepared from the degenerate oligonucleotide primers by PCR. Thus, every chromosome has a unique spectral color after in-situ hybridization with probes, which are differentially labelled with a mixture of fluorescent dyes (Rhodamine, Texas Red, Cy5, FITC and Cy5.5). The probes used for SKY consist of up to 55 chromosome specific probes 7-10.
The procedure for SKY involves several steps (Figure 1). SKY requires the availability of cells with high mitotic index from normal or diseased tissue or blood. The chromosomes of a single cell from either a freshly isolated primary cell or a cell line are spread on a glass slide. This chromosome spread is labeled with a different combination of fluorescent dyes specific for each chromosome. For probe detection and image acquisition,the spectral imaging system consists of sagnac interferometer and a CCD camera. This allows measurement of the visible light spectrum emitted from the sample and to acquire a spectral image from individual chromosomes. HiSKY, the software used to analyze the results of the captured images, provides an easy identification of chromosome anomalies. The end result is a metaphase and a karyotype classification image, in which each pair of chromosomes has a distinct color (Figure 2). This allows easy identification of chromosome identities and translocations. For more details, please visit Applied Spectral Imaging website (<a href="http://www.spectral-imaging.com/" target="_blank">http://www.spectral-imaging.com/</a>).
SKY was recently used for an identification of chromosome segregation defects and chromosome abnormalities in humans and mice with Autosomal Dominant Polycystic Kidney Disease (ADPKD), a genetic disease characterized by dysfunction in primary cilia 11-13. Using this technique, we demonstrated the presence of abnormal chromosome segregation and chromosomal defects in ADPKD patients and mouse models 14. Further analyses using SKY not only allowed us to identify chromosomal number and identity, but also to accurately detect very complex chromosomal aberrations such as chromosome deletions and translocations (Figure 2).

Spectral Karyotyping (SKY) is an advanced cytogenetics technique to identify genomic and chromosomal aberrations. This technique takes advantage of chromosome painting probes, which allow classification of all chromosomes. SKY can also identify complex chromosome aberrations and segregation defects in mice and humans with various diseases, including polycystic kidney disease.

多発性嚢胞腎疾患で、ヒトとマウスの染色体異常を研究するためにスペクトル核型分析

Conventional method to identify and classify individual chromosomes depends on the unique banding pattern of each chromosome in a specific species being ...

The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson's Disease

Parkinson's disease (PD) is the second most common movement disorder and affects 1% of people over the age of 60 1. Because ageing is the most important risk factor, cases of PD will increase during the next decades 2. Next to pathological protein folding and impaired protein degradation pathways, alterations of mitochondrial function and morphology were pointed out as further hallmark of neurodegeneration in PD 3-11.
After years of research in murine and human cancer cells as in vitro models to dissect molecular pathways of Parkinsonism, the use of human fibroblasts from patients and appropriate controls as ex vivo models has become a valuable research tool, if potential caveats are considered. Other than immortalized, rather artificial cell models, primary fibroblasts from patients carrying disease-associated mutations apparently reflect important pathological features of the human disease.
Here we delineate the procedure of taking skin biopsies, culturing human fibroblasts and using detailed protocols for essential microscopic techniques to define mitochondrial phenotypes. These were used to investigate different features associated with PD that are relevant to mitochondrial function and dynamics. Ex vivo, mitochondria can be analyzed in terms of their function, morphology, colocalization with lysosomes (the organelles degrading dysfunctional mitochondria) and degradation via the lysosomal pathway. These phenotypes are highly relevant for the identification of early signs of PD and may precede clinical motor symptoms in human disease-gene carriers. Hence, the assays presented here can be utilized as valuable tools to identify pathological features of neurodegeneration and help to define new therapeutic strategies in PD.

The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson&#039;s Disease

Fibroblasts from patients carrying mutations in Parkinson's disease-causing genes represent an easily accessible ex vivo model to study disease-associated phenotypes. Live cell imaging gives the opportunity to study morphological and functional parameters in living cells. Here we describe the preparation of human fibroblasts and subsequent monitoring of mitochondrial phenotypes.

パーキンソン病の分野におけるミトコンドリアの表現型を監視するための主要ヒト線維芽細胞の使用

Parkinson's disease (PD) is the second most  common movement disorder and affects 1% of people over the age of 60 1. Because ageing is  the most important ...

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery (SALLI) MRI in Rats

Small animal magnetic resonance imaging is an important tool to study cardiac function and changes in myocardial tissue. The high heart rates of small animals (200 to 600 beats/min) have previously limited the role of CMR imaging. Small animal Look-Locker inversion recovery (SALLI) is a T1 mapping sequence for small animals to overcome this problem 1. T1 maps provide quantitative information about tissue alterations and contrast agent kinetics. It is also possible to detect diffuse myocardial processes such as interstitial fibrosis or edema 1-6. Furthermore, from a single set of image data, it is possible to examine heart function and myocardial scarring by generating cine and inversion recovery-prepared late gadolinium enhancement-type MR images 1.
	The presented video shows step-by-step the procedures to perform small animal CMR imaging. Here it is presented with a healthy Sprague-Dawley rat, however naturally it can be extended to different cardiac small animal models.

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery SALLI MRI in Rats

Contrast enhanced cardiac magnetic resonance (CMR) imaging allows comprehensive in vivo assessment of the heart in small animal models of cardiovascular disease. Here we detail the procedures to perform CMR imaging, reconstruction and analysis using Small Animal Lock-Locker Inversion Recovery (SALLI) in rats.

小動物ルックロッカー反転回復（サリー）ラットにおけるMRIを用いた心機能と心筋形態の評価

Small animal magnetic resonance  imaging is an important tool to study cardiac function and changes in myocardial  tissue. The high heart rates of small ...

In vivo 19F MRI for Cell Tracking

In vivo 19F MRI allows quantitative cell tracking without the use of ionizing radiation. It is a noninvasive technique that can be applied to humans. Here, we describe a general protocol for cell labeling, imaging, and image processing. The technique is applicable to various cell types and animal models, although here we focus on a typical mouse model for tracking murine immune cells. The most important issues for cell labeling are described, as these are relevant to all models. Similarly, key imaging parameters are listed, although the details will vary depending on the MRI system and the individual setup. Finally, we include an image processing protocol for quantification. Variations for this, and other parts of the protocol, are assessed in the Discussion section. Based on the detailed procedure described here, the user will need to adapt the protocol for each specific cell type, cell label, animal model, and imaging setup. Note that the protocol can also be adapted for human use, as long as clinical restrictions are met.

In vivo 19F MRI for Cell Tracking

We describe a general protocol for in vivo cell tracking using MRI in a mouse model with ex vivo labeled cells. A typical protocol for cell labeling, image acquisition processing and quantification is included.

細胞追跡のためのin vivoでの19 F MRI における

In vivo 19F MRI allows quantitative cell tracking without the use of ionizing radiation. It is a noninvasive technique that can be applied to humans. ...

MRI and PET in Mouse Models of Myocardial Infarction

Myocardial infarction is one of the leading causes of death in the Western world. The similarity of the mouse heart to the human heart has made it an ideal model for testing novel therapeutic strategies.
In vivo magnetic resonance imaging (MRI) gives excellent views of the heart noninvasively with clear anatomical detail, which can be used for accurate functional assessment. Contrast agents can provide basic measures of tissue viability but these are nonspecific. Positron emission tomography (PET) is a complementary technique that is highly specific for molecular imaging, but lacks the anatomical detail of MRI. Used together, these techniques offer a sensitive, specific and quantitative tool for the assessment of the heart in disease and recovery following treatment.
In this paper we explain how these methods are carried out in mouse models of acute myocardial infarction. The procedures described here were designed for the assessment of putative protective drug treatments. We used MRI to measure systolic function and infarct size with late gadolinium enhancement, and PET with fluorodeoxyglucose (FDG) to assess metabolic function in the infarcted region. The paper focuses on practical aspects such as slice planning, accurate gating, drug delivery, segmentation of images, and multimodal coregistration. The methods presented here achieve good repeatability and accuracy maintaining a high throughput.

We describe how to perform MRI and PET imaging of the mouse heart. The protocol is tailored to assess treatment efficacy in models of myocardial infarction and heart failure.

Myocardial infarction is one of the leading causes of death in the Western world. The similarity of the mouse heart to the human heart has made it an ...

A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys

Polycystic kidney disease (PKD) is one of the most common causes of end-stage kidney disease, a devastating disease for which there is no cure. The molecular mechanisms leading to cyst formation in PKD remain somewhat unclear, but many genes are thought to be involved. Wnt5a is a non-canonical glycoprotein that regulates a wide range of developmental processes. Wnt5a works through the planar cell polarity (PCP) pathway that regulates oriented cell division during renal tubular cell elongation. Defects of the PCP pathway have been found to cause kidney cyst formation. Our paper describes a method for developing a zebrafish cystic kidney disease model by knockdown of the wnt5a gene with wnt5a antisense morpholino (MO) oligonucleotides. Tg(wt1b:GFP) transgenic zebrafish were used to visualize kidney structure and kidney cysts following wnt5a knockdown. Two distinct antisense MOs (AUG - and splice-site) were used and both resulted in curly tail down phenotype and cyst formation after wnt5a knockdown. Injection of mouse Wnt5a mRNA, resistant to the MOs due to a difference in primary base pair structure, rescued the abnormal phenotype, demonstrating that the phenotype was not due to &#8220;off-target&#8221; effects of the morpholino. This work supports the validity of using a zebrafish model to study wnt5a function in the kidney.

A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys

We describe a method of generating a possible zebrafish model of polycystic kidney disease. We used Tg(wt1b:GFP) fish to visualize kidney structure. Knockdown of wnt5a was by morpholino injection. Pronephric cyst formation after wnt5a knockdown was observed in this GFP transgenic zebrafish.

多発性嚢胞腎疾患の可能性のあるゼブラフィッシュモデル：のノックダウン<em&gt; WNT5A</em&gt;ゼブラフィッシュ腎臓で嚢胞を引き起こす

Polycystic kidney disease (PKD) is one of the most common causes of end-stage kidney disease, a devastating disease for which there is no cure. The ...

Ex Vivo Intestinal Sacs to Assess Mucosal Permeability in Models of Gastrointestinal Disease

The epithelial barrier is the first innate defense of the gastrointestinal tract and selectively regulates transport from the lumen to the underlying tissue compartments, restricting the transport of smaller molecules across the epithelium and almost completely prohibiting epithelial macromolecular transport. This selectivity is determined by the mucous gel layer, which limits the transport of lipophilic molecules and both the apical receptors and tight junctional protein complexes of the epithelium. In vitro cell culture models of the epithelium are convenient, but as a model, they lack the complexity of interactions between the microbiota, mucous-gel, epithelium and immune system. On the other hand, in vivo assessment of intestinal absorption or permeability may be performed, but these assays measure overall gastrointestinal absorption, with no indication of site specificity. Ex vivo permeability assays using &#34;intestinal sacs&#34; are a rapid and sensitive method of measuring either overall intestinal integrity or comparative transport of a specific molecule, with the added advantage of intestinal site specificity. Here we describe the preparation of intestinal sacs for permeability studies and the calculation of the apparent permeability (Papp) of a molecule across the intestinal barrier. This technique may be used as a method of assessing drug absorption, or to examine regional epithelial barrier dysfunction in animal models of gastrointestinal disease.

Ex Vivo Intestinal Sacs to Assess Mucosal Permeability in Models of Gastrointestinal Disease

This protocol describes the use of excised intestinal tissue preparations or "intestinal sacs" as an ex vivo model of intestinal barrier function. This model may be used to assess integrity of both the epithelial barrier and the mucous gel layer at specific intestinal sites in animal models of digestive disease.

ex vivoで腸嚢は、胃腸疾患のモデルに粘膜透過性を評価するために

The epithelial barrier is the first innate defense of the gastrointestinal tract and selectively regulates transport from the lumen to the underlying ...

Quantitative 3D In Silico Modeling (q3DISM) of Cerebral Amyloid-beta Phagocytosis in Rodent Models of Alzheimer's Disease

Neuroinflammation is now recognized as a major etiological factor in neurodegenerative disease. Mononuclear phagocytes are innate immune cells responsible for phagocytosis and clearance of debris and detritus. These cells include CNS-resident macrophages known as microglia, and mononuclear phagocytes infiltrating from the periphery. Light microscopy has generally been used to visualize phagocytosis in rodent or human brain specimens. However, qualitative methods have not provided definitive evidence of in vivo phagocytosis. Here, we describe quantitative 3D in silico modeling (q3DISM), a robust method allowing for true 3D quantitation of amyloid-&#946; (A&#946;) phagocytosis by mononuclear phagocytes in rodent Alzheimer&#39;s Disease (AD) models. The method involves fluorescently visualizing A&#946; encapsulated within phagolysosomes in rodent brain sections. Large z-dimensional confocal datasets are then 3D reconstructed for quantitation of A&#946; spatially colocalized within the phagolysosome. We demonstrate the successful application of q3DISM to mouse and rat brains, but this methodology can be extended to virtually any phagocytic event in any tissue.

Quantitative 3D In Silico Modeling q3DISM of Cerebral Amyloid-beta Phagocytosis in Rodent Models of Alzheimer&#039;s Disease

We developed a methodology for quantitative 3D in silico modeling (q3DISM) of cerebral amyloid-&#946; (A&#946;) phagocytosis by mononuclear phagocytes in rodent models of Alzheimer&#39;s disease. This method can be generalized for the quantitation of virtually any phagocytic event in vivo.

定量的3D<em&gt;インシリコ</emアルツハイマー病の齧歯類モデルにおける脳アミロイドベータ食作用の&gt;モデリング（q3DISM）

Neuroinflammation is now recognized as a major etiological factor in neurodegenerative disease. Mononuclear phagocytes are innate immune cells responsible ...

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and &#181;CT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

The evaluation of tissue development in the fracture callus during endochondral bone healing is essential to monitor the healing process. Here, we report the use of a magnetic resonance imaging (MRI)-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice.

生体内で破壊カルス開発マウス大腿骨の骨、MRI 対応のデバイスを用いたマウスの骨治癒過程の評価

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...