Name: use of ultra-high field mri in small rodent models of polycystic kidney disease for in vivo phenotyping and drug monitoring
Uploaded: 2015-06-23T14:00:00.000+00:00
Duration: 7 min 35 s
Description: 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

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).

Canlı hayvan ile herhangi prosedürleri başlamadan önce, deney protokolleri kurumsal hayvan bakımı ve kullanımı komitesi (IACUC) tarafından onaylanmış olması gerekir. 1. Tarayıcı Yapılandırması <ol><li> Başlamadan önce, ısıtıcı KAPALI konumda olduğundan emin olun. </li><li> Mini görüntüleme degrade ve 38 mm RF bobini mini görüntüleme tutucu seçin. </li><li> Sahibinin merkez deliğinde değişken sıcaklık düzeneğini takın. </li></ol> 2. Hayvan hazırlanması <ol><li> MR deneyler için, buharlaşmış izofluran kullanarak optimum anestezi elde. Anestezinin indüksiyonu için, bir emici doku ile kaplı bir indüksiyon odasında hayvan. 2.0-2.5 L / dk ve oksijen% 3 izofluran izofluran buharlaştırıcı debi ölçer ayarlayın. </li><li> Bu aşamada herhangi bir metal etiketi veya diğer metalik nesneyi kaldırın. Anestezi altında iken kuruluğunu önlemek için hayvanın gözleri veteriner merhem sürün. </li><li> Hayvan kezanestezi, cerrahi düzlemine ulaşmıştır (yani, ayak tutam çekilme refleks kaybı), bir burun konisi içine yerleştirilir onun burunlu bir tutucuya hayvan yerleştirin. 2.0-2.5 ml / dk soruşturmada anestezi hava akımını ve oksijen 1.5-2.0% izofluran konsantrasyonu ayarlayın. Anestezi işlemi sırasında burun konisi ile teslim edilecektir. Periyodik 40 bpm ~ bir solunum hızını korumak için hayvanın yaşına ve ağırlığına bağlı olarak izofluran konsantrasyonu ayarlayın. </li><li> Yerine hayvan sabitlemek ve MRG deney sırasında hareket etmesini önlemek için hayvan sahiplerine kullanın. Vücut bölgesine bağlı olarak hayvan sahibinin tipi taranacak Vary. Not: Laboratuvar plastik (polipropilen, Teflon, polistren, polikarbonat) den Özelleştirilmiş sahipleri belirli deney karşılamak ve (160 gr sıçan yenidoğan fareden) hayvan boyutuna uyacak şekilde yapılabilir. </li><li> Hayvanın vücut sıcaklığını izlemek için hayvanda rektal termometre yerleştirin. Exp sırasındaeriment, bir sıcak hava akımı kullanılarak, 35-37 ° C&#39;de hayvan tutun. Hava sıcaklığı (30-38 ° C) ve hayvanın vücut ısısı yorumlarına dayanarak akışı (1,200-2,000 L / saat) ayarlayın. </li><li> Solunum hızını izlemek için hayvanın karın bir balon solunum basınç sensörü takın. </li><li> RF bobini merkezinde hayvan Güvenli ve dikkatli MRI tarayıcısı içine hayvan ile RF bobin yerleştirin. </li></ol> 3. MR Deney <ol><li> Ayarlama deneyleri RF gücü kullanılan en aza indirmek için ve sinyal-gürültü oranının en üst düzeye çıkarmak için başlamadan önce RF bobini maç ve. Eşleştirme / ayarlamayı başlatmak için: <ol><li> Araçlar simgesini tıklatarak spektrometre kontrol aracını açın. </li><li> Spektrometre kontrol aracı olarak Acquisition → yalpalama tıklatın. Bir Edi / Reco penceresi yalpalama eğrisi görüntüleyen açılacaktır. </li><li> Alternatif yansıyan RF gücü kadar küçük adımlarla ayarlama ve eşleştirme kapasitörler (çubuklar ayar kullanarak ve eşleştirme) ayarlamaken aza indirilir. amaç yatay eksende sıfır konumlandırılmış dikey eksende minimum bir eğri görmektir. </li><li> Bobinin kalibrasyon başarıyla elde edildiğinde, Edi / Reco penceresindeki Durdur düğmesine basın. </li></ol></li><li> Eksenel koronal ve sagital görüntüler oluşturmak için üç ortogonal düzlemde izci görüntü elde. Izci görüntüler elde etmek için 12 gibi içi Kapısı Hızlı Düşük Açılı Çekim (IG-FLASH) gibi hızlı görüntü dizisi kullanın. Gerçek görüntüleme için uygun geometriyi ayarlamak için keşif görüntüleri kullanın. </li><li> Özel araştırma amaçlarına bağlı olarak, uygun görüntü dizisi ve parametreleri seçin ve bir trafik ışığı ile tarama başlatın. Bu, tüm otomatik, RF kanalı kalibre su üzerinde rezonans mıknatıs, set taşıyıcı frekansı şim ve alıcı kazancı ayarlar. <ol><li> Anatomik çalışmalar ve T2 ağırlıklı görüntülerde, 2B çoklu dilim veya 3D modunda kazanır. Alan tutmak, belirli bir uzamsal çözünürlük için deney süresini kısaltmak için-Tabii-view (FOV) mümkün ama wrap-around eserler (2,56-3,2 cm) önlemek için yeterince büyük olduğu kadar küçük. </li></ol></li><li> Tekrarlama süresi (TR) ve / veya dilim sayısı uygun bir şekilde seçilmesi ile, hayvan respirasyon döngüsü biraz daha kısa seçilen sekansın döngüsü tutun. Bu veriler hayvanların sessiz dönemde tahsil edilmesini sağlar. <ol><li> Örneğin, karın görüntüler için, ~ 30 bpm hayvanın solunum hızı tutmak; Bu nefes başına yaklaşık 2,000 milisaniye olduğunu. TR, Dinlenme Geliştirme (NADİR) dizisi ile Turbo Hızlı Toplama kullanın ve 11-19 koronal dilim elde / TE 1500/9 msn, NADİR faktör 8 ve (matris 256 x 256, FOV 2.56 x 2.56 cm, dilim kalınlığı 0.75 mm) . Not: 1500 msn TR ayarlayarak ve ~ 30 bpm (nefes başına 2,000 msn) hayvanın solunum hızı tutarak, biz veri hayvanların sessiz dönemde toplanan emin olun. </li></ol></li><li> Tüm görüntü elde etme tamamlandıktan sonra, taranan hayvan koyunve ısıtmalı pad üzerinde ayaktan kadar izler. İyileştikten sonra, kafesine hayvan dönün ve hayvan tesis dönmeden önce 1 saat en az izlenen. </li></ol>

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The authors have nothing to disclose.

Bu yazıda, in vivo fenotipik karakterizasyonu veya PKD için kemirgen modellerinde ilaç izleme için bir araç olarak UHF MRI kullanılarak fizibilitesini göstermektedir. Biz geniş bir delik mikro ve küçük görüntüleme aksesuarlarla donatılmış Avance III yüksek çözünürlüklü NMR spektrometre ile 16.4 T yapılan deneyler açıklar. spektrometre Paravision 5.1 görüntüleme yazılımı tarafından kontrol TopSpin2.0PV toplama ve işleme yazılımı tarafından tahri...

Polikistik Böbrek Hastalığı (PKD), böbrek kist gelişimi ile karakterize tek gen hastalıkları grubunu içerir. Aralarında Otozomal dominant polikistik böbrek hastalığı (ADPKD) ve otozomal resesif polikistik böbrek hastalığı en yaygın türleri 1,2 temsil (ORPBH) bulunmaktadır. Polikistik böbrek, kalıtsal böbrek kistik hastalıkların en sık görülen formu, PKD1 veya PKD2 genlerindeki mutasyonlarla kaynaklanmaktadır. Geç başlangıçlı, birden iki taraflı böbrek kistleri, değişken ekstra renal kistler eşlik yanı sıra kardiyovasküler ve kas iskelet anomalileri ile karakterizedir. ORPBH, en sık etkileyen yenidoğan ve küçük çocuklar, PKHD1 mutasyonlar neden olur ve genişlemiş ekojenik böbrek ve konjenital hepatik fibrozis, 3 ile karakterizedir. Önemli bir şekilde, polikistik böbrek hem büyük p sonuçlanan bir genin (gen) ve mutasyon (alelik) seviyesi, en heterojenlik ile karakterize edilirhenotypic değişkenliği. PKD1 genindeki mutasyonlar ağır klinik tablo (sayısız kistler, erken tanı, hipertansiyon ve hematüri) yanı sıra, (PKD2 mutasyonları olan hastalardan daha 20 yıl önce) böbrek hastalığı son dönem hızla ilerlemesi ile ilişkili 4. Ağır polikistik karaciğer hastalığı (PLD) ve vasküler anormallikler PKD1 ve PKD2 5, her iki mutasyon ile ilişkili olabilir. bu hastalığın renal komplikasyonlara çoğunluğu ilişkili enflamasyon ve fibroz ile birlikte kist genişleme bir sonucu olarak özellikle ortaya çıkmaktadır. Kist geliştirme intrauterin başlar ve hasta ömür boyu devam eder. Böbrekler genellikle 20&#39;den fazla kez normal böbrek hacmi ulaşabilir bile onların reniform şeklini korumak. Böbrek kistlerinin hastalar mevcut ikili dağıtım çoğu, ancak bazı sıradışı durumlarda, kist tek taraflı veya asimetrik desen gelişebilir. Önemli bir Challennefrologlar Otozomal dominant polikistik böbrek hastaları takip veya tedaviler uygulanması için ge hastalığın doğal seyri olduğunu. Onun en tabii sırasında böbrek fonksiyonları normale kalır ve böbrek fonksiyonu azalmaya başlar zaman, böbrekler çoğu kistler almıştır. Terapiler sonraki aşamalarda uygulandığında, hastanın zaten kronik böbrek hastalığında geri dönüşü olmayan bir noktaya ulaşmış olabilir çünkü başarılı olmak için daha az olasıdır. Tedavi, erken aşamalarında başlatılan Aksine, bu sadece glomerüler filtrasyon hızı dayanan bir tepki tespit etmek güçtür. Bunun bir sonucu olarak, hastalık ilerlemesinin bir göstergesi olarak, böbrek hacminin kavramı dikkat çekti. Otozomal dominant polikistik böbrek hastalarında böbrek ve kist hacmindeki artış, doğrudan olarak toplam Böbrek Hacim (TKV) potansiyelini vurgulayan, böbrek fonksiyon bozukluğu ile ilişkili olduğunu Polikistik Böbrek Hastalığı (CRISP) Çalışmanın Radyolojik Görüntüleme Çalışmaları Konsorsiyumu göstermiştirhastalığın ilerlemesi 6,7 için urrogate işaretleyici. Sonuç olarak, şu anda TKV ADPKD 2,8,9 çoklu klinik çalışmalarda birincil ya da ikincil bir son nokta olarak kullanılır. Birden fare modelleri de dahil olmak üzere spontan mutasyonlar ve genetik olarak PKD 10,11 patogenezinde ışık tutmuştur. PKD1 veya PKD2 modelleri (PKD1 veya PKD2 ya mutasyonlar), onlar mükemmel taklit insan hastalığı en popüler olanları haline gelmiştir. Buna ek olarak, PKD1 ya PKD2 genler dışında genlerdeki mutasyonlarla kemirgen modelleri hastalığı ile ilişkili sinyal yollarının aydınlatmak için deneysel bir platform olarak kullanılmıştır. Buna ek olarak, bu modellerin muhtelif tedaviler test etmek için kullanılmıştır. Ancak, PKD birçok kemirgen çalışmalarında bir sınırlayıcı faktör sırasıyla böbrek anatomik ve fonksiyonel değişiklikleri analiz etmek için etkili non-invaziv yöntemlerin eksikliği genellikle. Manyetik resonance görüntüleme (MRG) mükemmel yumuşak doku kontrastı ve anatomik detay sağlayan ve TKV ölçümleri sağlayan ADPKD hastaları takip etmek Geçerli altın standart görüntüleme tekniğidir. MRG de in vivo olarak büyük hayvan ve insanlarda, görüntüleme küçük kemirgenler anatomik görüntüleme için kurulmuş olsa bile yüksek çözünürlüklü görüntüleri elde etmek için yeteneği yararlılığını kısıtlayabilir ek teknik zorlukları, gerektirir. Ultra yüksek alan (UHF) MR (7-16,4 T) ve daha güçlü geçişlerini gelişimi tanıtımı ile, benzer gürültü oranları ve teşhis kalitesi ile MR görüntülerinin mekansal çözünürlükte sinyal yüksek ulaşmak artık mümkün insanlarda elde edilmiştir. Sonuç olarak, PKD küçük kemirgen modellerinde in vivo görüntüleme için UHF MRI kullanımı araştırmacılar için güçlü bir araç haline gelmiştir.

<table><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 &lt;em&gt;in vivo&lt;/em&gt; small rodent imaging</td></tr><tr><td>TopSpin2.0PV</td><td>Bruker</td><td>H9088TA2</td><td>Spectrometer processing software</td></tr><tr><td>Paravision 5.1</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></tr></tbody></table>

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.

Bu yazıda, PKD ve diğer böbrek hastalıkları için kemirgen modellerinde in vivo fenotipik karakterizasyonu veya ilaç takibi için bir araç olarak UHF MR yararlılığını göstermeyi hedeflemekteyiz. Deneyler Tüm IACUC tarafından onaylanmış deneysel protokollerin bir parçası idi. PKD UHF MRI kullanarak küçük kemirgen modellerinde in vivo feno: Tüm görüntüleme çalışmaları Bruker AVANCEIII-700 (16.4 T) ile, izofluran anestezi...

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.

Için Polikistik Böbrek Hastalığında Küçük Kemirgen modelleri Ultra yüksek Saha MRI Kullanımı<em&gt; In Vivo</em&gt; Fenotiplendirme ve İlaç İzleme

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

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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 Görüntüleme.  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.

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

Utilizing High Resolution Ultrasound to Monitor Tumor Onset and Growth in Genetically Engineered Pancreatic Cancer Models

The LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre (KPC) mouse model represents an established and frequently used transgenic model to evaluate novel therapies in pancreatic cancer. Tumor onset is variable in the KPC model between 8 weeks and several months. Therefore, non-invasive imaging tools are required to screen for tumor onset and monitor for response to treatment. To address this issue, different approaches have emerged over the last years. High resolution ultrasound has major advantages such as non-invasiveness, fast session times and a high image resolution without radiation exposure. However, ultrasound in mice is not trivial and sufficient anatomical knowledge and practical skills are required to successfully perform high resolution ultrasound in preclinical pancreatic cancer models. With the following article, a detailed hands-on guide for abdominal ultrasound in murine models with a particular focus on endogenous pancreatic cancer models is presented. Furthermore, a summary of common mistakes and how to avoid them is provided.

This article describes the utilization of high-resolution ultrasound in genetically engineered pancreatic cancer mice. The primary aim is to provide a detailed instruction for detection and evaluation of endogenous pancreatic tumors.

Yüksek çözünürlüklü ultrason monitör tümör başlangıçlı ve büyüme genetiği pankreas kanseri modellerinde kullanmak

The LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre (KPC) mouse model represents an established and frequently used transgenic model to evaluate novel ...

Kanser Araştırmaları

Multiple-mouse Neuroanatomical Magnetic Resonance Imaging

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and evaluating mouse models of human disease. MRI is an excellent modality for investigating genetically altered animals. It is capable of whole brain coverage, can be used in vivo, and provides multiple contrast mechanisms for investigating different aspects of neuranatomy and physiology. The advent of high-field scanners along with the ability to scan multiple mice simultaneously allows for rapid phenotyping of novel mutations.
Effective mouse MRI studies require attention to many aspects of experiment design. In this article, we will describe general methods to acquire quality images for mouse phenotyping using a system that images mice concurrently in shielded transmit/receive radio frequency (RF) coils in a common magnet (Bock et al., 2003). We focus particularly on anatomical phenotyping, an important and accessible application that has shown a high potential for impact in many mouse models at our imaging centre. Before we can provide the detailed steps to acquire such images, there are important practical considerations for both in vivo brain imaging (Dazai et al., 2004) and ex vivo brain imaging (Spring et al., 2007) that should be noted. These are discussed below.

Magnetic resonance imaging (MRI) has become an increasingly popular tool for examining the phenotype of genetically altered mice. This article illustrates the methods necessary to achieve high-throughput phenotyping of genetically altered mice using multiple-mouse MRI.

Çoklu-fare nöroanatomik Manyetik Rezonans Görüntüleme

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and ...

Nörobilim

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.

Polikistik Böbrek Hastalığı Olan İnsanlar ve Fareler kromozom anomalileri Eğitim Spektral Karyotipik

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

Tıp

Early Detection of Drug-Induced Renal Hemodynamic Dysfunction Using Sonographic Technology in Rats

The kidney normally functions to maintain hemodynamic homeostasis and is a major site of damage caused by drug toxicity. Drug-induced nephrotoxicity is estimated to contribute to 19- 25% of all clinical cases of acute kidney injury (AKI) in critically ill patients. AKI detection has historically relied on metrics such as serum creatinine (sCr) or blood urea nitrogen (BUN) which are demonstrably inadequate in full assessment of nephrotoxicity in the early phase of renal dysfunction. Currently, there is no robust diagnostic method to accurately detect hemodynamic alteration in the early phase of AKI while such alterations might actually precede the rise in serum biomarker levels. Such early detection can help clinicians make an accurate diagnosis and help in in decision making for therapeutic strategy. Rats were treated with Cisplatin to induce AKI. Nephrotoxicity was assessed for six days using high-frequency sonography, sCr measurement and upon histopathology of kidney. Hemodynamic evaluation using 2D and Color-Doppler images were used to serially study nephrotoxicity in rats, using the sonography. Our data showed successful drug-induced kidney injury in adult rats by histological examination. Color-Doppler based sonographic assessment of AKI indicated that resistive-index (RI) and pulsatile-index (PI) were increased in the treatment group; and peak-systolic velocity (mm/s), end-diastolic velocity (mm/s) and velocity-time integral (VTI, mm) were decreased in renal arteries in the same group. Importantly, these hemodynamic changes evaluated by sonography preceded the rise of sCr levels. Sonography-based indices such as RI or PI can thus be useful predictive markers of declining renal function in rodents. From our sonography-based observations in the kidneys of rats that underwent AKI, we showed that these noninvasive hemodynamic measurements may consider as an accurate, sensitive and robust method in detecting early stage kidney dysfunction. This study also underscores the importance of ethical issues associated with animal use in research.

Early stage hemodynamic dysfunction is critical to the development of kidney disease. Yet, detection methodologies are limited. Recent advances in sonography provide a noninvasive, accurate option for early detection of kidney injury. This study outlines a step-by-step, sonographic methodology for detecting kidney dysfunction using a drug-induced nephrotoxicity rat model.

Sıçanlarda sonografik Teknoloji Kullanımı İlaç Kaynaklı Böbrek Hemodinamik Bozukluğu Erken Tespiti

The kidney normally functions to maintain hemodynamic homeostasis and is a major site of damage caused by drug toxicity. Drug-induced nephrotoxicity is ...

Transcutaneous Assessment of Renal Function in Conscious Rodents

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and time-consuming. In addition, serial blood or urine samples are required, with the associated stress for the experimental animals. A recent technique significantly reduces the investment in time and resources, minimizing the invasiveness and the animal stress, but being equally valid as the traditional approaches. The method measures transcutaneously renal function. Using an optical device and the exogenous renal marker fluorescein isothiocyanate (FITC)-sinistrin, this technique is capable of measuring the elimination kinetics of the marker through the skin. With neither blood nor urine samples nor the associated laboratory assays needed, the results of the transcutaneous measurement are almost instantaneously available. The method has been already validated in different species and successfully applied in several models of renal pathology. Moreover, due to its minimally invasive characteristics, it is suitable for sequential measurements within the same animal. Here is provided a detailed protocol to carry out the transcutaneous assessment of renal function in rodents.

Determination of glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional procedures to measure this parameter are cumbersome and require a large investment of time. Here we describe a faster and minimally invasive method to determinate GFR transcutaneously.

Bilinçli Kemirgenler Böbrek Fonksiyon Transkutanöz Değerlendirmesi

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and ...

Assessment of Kidney Function in Mouse Models of Glomerular Disease

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function in diabetic nephropathy and podocyte-specific VEGF-A knock-out mice; therefore, this protocol describes the full kidney work-up used in our lab to assess these mouse models of glomerular disease, enabling a vast amount of information regarding kidney and glomerular function to be obtained from a single mouse. In comparison to alternative methods presented in the literature to assess glomerular function, the use of the method outlined in this paper enables the glomerular phenotype to be fully evaluated from multiple aspects. By using this method, the researcher can determine the kidney phenotype of the model and assess the mechanism as to why the phenotype develops. This vital information on the mechanism of disease is required when examining potential therapeutic avenues in these models. The methods allow for detailed functional assessment of the glomerular filtration barrier through measurement of the urinary albumin creatinine ratio and individual glomerular water permeability, as well as both structural and ultra-structural examination using the Periodic Acid Schiff stain and electron microscopy. Furthermore, analysis of the genes dysregulated at the mRNA and protein level enables mechanistic analysis of glomerular function. This protocol outlines the generic but adaptable methods that can be applied to all mouse models of glomerular disease.

This protocol describes a full kidney work-up that should be carried out in mouse models of glomerular disease. The methods allow for detailed functional, structural, and mechanistic analysis of glomerular function, which can be applied to all mouse models of glomerular disease.

Fare modelleri Glomerular hastalığı böbrek fonksiyonlarının değerlendirilmesi

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function ...

Use of 3D Robotic Ultrasound for In Vivo Analysis of Mouse Kidneys

Common modalities for in vivo imaging of rodents include positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), and ultrasound (US). Each method has limitations and advantages, including availability, ease of use, cost, size, and the use of ionizing radiation or magnetic fields. This protocol describes the use of 3D robotic US for in vivo imaging of rodent kidneys and heart, subsequent data analysis, and possible research applications. Practical applications of robotic US are the quantification of total kidney volume (TKV), as well as the measurement of cysts, tumors, and vasculature. Although the resolution is not as high as other modalities, robotic US allows for more practical high throughput data collection. Furthermore, using US M-mode imaging, cardiac function may be quantified. Since the kidneys receive 20%-25% of the cardiac output, assessing cardiac function is critical to the understanding of kidney physiology and pathophysiology.

Use of 3D Robotic Ultrasound for In Vivo Analysis of Mouse Kidneys

This protocol demonstrates robotic ultrasound (US) as a practical, cost-effective, and quick alternative to traditional non-invasive image modalities.

Fare Böbreklerinin In Vivo Analizi için 3D Robotik Ultrason Kullanımı

Common modalities for in vivo imaging of rodents include positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), ...

Using 2-Photon Microscopy to Quantify the Effects of Chronic Unilateral Ureteral Obstruction on Glomerular Processes

Applying novel microscopy methods to suitable animal disease models to explore the dynamic physiology of the kidney remains a challenge. Rats with surface glomeruli provide a unique opportunity to investigate physiological and pathophysiological processes using intravital 2-photon microscopy. Quantification of glomerular capillary blood flow and vasoconstriction and dilatation in response to drugs, permeability, and inflammation are just some of the processes that can be studied. In addition, transgenic rats, i.e., podocytes labeled with fluorescent dyes and other molecular biomarker approaches, provide increased resolution to directly monitor and quantify protein-protein interactions and the effects of specific molecular alterations.
In mice, which lack surface glomeruli after four weeks of age, unilateral ureteral obstruction (UUO) for several weeks has been used to induce surface glomeruli. As this induction model does not allow for baseline studies, we quantified the effects of UUO on glomerular processes in the UUO model in Munich Wistar Fr&#246;mter (MWF) rats, which have surface glomeruli under physiologic conditions. The UUO model for five weeks or more induced significant alterations to gross renal morphology, the peritubular and glomerular microvasculature, as well as the structure and function of tubular epithelia. Glomerular and peritubular red blood cell (RBC) flow decreased significantly (p &#60; 0.01), probably due to the significant increase in the adherence of white blood cells (WBCs) within glomerular and peritubular capillaries. The glomerular sieving coefficient of albumin increased from 0.015 &#177; 0.002 in untreated MWFs to 0.045 &#177; 0.05 in 5-week-old UUO MWF rats. Twelve weeks of UUO resulted in further increases in surface glomerular density and glomerular sieving coefficient (GSC) for albumin. Fluorescent albumin filtered across the glomeruli was not reabsorbed by the proximal tubules. These data suggest that using UUO to induce surface glomeruli limits the ability to study and interpret normal glomerular processes and disease alterations.

Here, we present a protocol using 2-photon microscopy in Munich Wistar Fromter rats with surface glomeruli to quantifythe effects of prolonged ureteral obstruction on glomerular dynamics and function.

Kronik Tek Taraflı Üreter Tıkanıklığının Glomerüler Prosesler Üzerindeki Etkilerini Ölçmek için 2-Foton Mikroskobu Kullanılması

Applying novel microscopy methods to suitable animal disease models to explore the dynamic physiology of the kidney remains a challenge. Rats with surface ...

High-Resolution Cardiac Positron Emission Tomography/Computed Tomography for Small Animals

Positron emission tomography (PET) and computed tomography (CT) are among the most employed diagnostic imaging techniques, and both serve in understanding cardiac function and metabolism. In preclinical research, dedicated scanners with high sensitivity and high spatio-temporal resolution are employed, designed to cope with the demanding technological requirements posed by the small heart size and very high heart rates of mice and rats. In this paper, a bimodal cardiac PET/CT imaging protocol for experimental mouse and/or rat models of cardiac diseases is described, from animal preparation and image acquisition and reconstruction to image processing and visualization.
In particular, the 18F-labeled fluorodeoxyglucose ([18F]FDG)-PET scan allows for the measurement and visualization of glucose metabolism in the different segments of the left ventricle (LV). Polar maps are convenient tools to display this information. The CT part consists of a time-resolved 3D reconstruction of the entire heart (4D-CT) using retrospective gating without electrocardiography (ECG) leads, allowing the morphofunctional evaluation of the LV and the subsequent quantification of the most important cardiac function parameters, such as ejection fraction (EF) and stroke volume (SV). Using an integrated PET/CT scanner, this protocol can be executed within the same anesthesia induction without the need to reposition the animal between different scanners. Hence, PET/CT can be seen as a comprehensive tool for the morphofunctional and metabolic evaluation of the heart in several small animal models of cardiac diseases.

Here, we present an experimental imaging protocol for the quantification of cardiac function and morphology using high-resolution positron emission tomography/computed tomography for small animals. Both mice and rats are considered, discussing the different requirements of computed tomography contrast agents for the two species.

Yüksek Çözünürlüklü Kardiyak Pozitron Emisyon Tomografisi/Küçük Hayvanlar için Bilgisayarlı Tomografi

Positron emission tomography (PET) and computed tomography (CT) are among the most employed diagnostic imaging techniques, and both serve in understanding ...

Ultrafast Doppler Ultrasound for Hepatic Microvasculature Analysis in Mice

Imaging and Quantification of the Hepatic Vasculature of Mice Using Ultrafast Doppler Ultrasound

Non-invasive in vivo imaging of the vasculature is a powerful tool for studying disease mechanisms in rodents. To achieve high sensitivity imaging of the microvasculature using Doppler ultrasound methods, imaging modalities employing the concept of ultrafast imaging are preferred. By increasing the frame rate of the ultrasound scanner to thousands of frames per second, it becomes possible to improve the sensitivity of the blood flow down to 2 mm/s and to obtain functional information about the microcirculation in comparison to a sensitivity of around 1 cm/s in conventional Doppler modes. While Ultrafast Doppler ultrasound (UFUS) imaging has become adopted in neuroscience, where it can capture brain activity through neurovascular coupling, it presents greater challenges when imaging the vasculature of abdominal organs due to larger motions linked to breathing. The liver, positioned anatomically under the diaphragm, is particularly susceptible to out-of-plane movement and oscillating respiratory motion. These artifacts not only adversely affect Doppler imaging but also complicate the anatomical analysis of vascular structures and the computation of vascular parameters. Here, we present a qualitative and quantitative imaging analysis of the hepatic vasculature in mice by UFUS. We identify major anatomical vascular structures and provide graphical illustrations of the hepatic macroscopical anatomy, comparing it to an in-depth anatomical assessment of the hepatic vasculature based on Doppler readouts. Additionally, we have developed a quantification protocol for robust measurements of hepatic blood volume of the microvasculature over time. To contemplate further research, qualitative vascular analysis provides a comprehensive overview and suggests a standardized terminology for researchers working with mouse models of liver disease. Furthermore, it offers the opportunity to apply ultrasound as a non-invasive complementary method to inspect hepatic vascular defects in vivo and measure functional microvascular alterations deep within the organ before unraveling blood vessel anomalies at the micron scale levels using ex vivo staining on tissue sections.

Ultrafast Doppler ultrasound (UFUS) with high spatial resolution (100 &#956;m) and sensitivity represents a suitable noninvasive imaging modality for obtaining a comprehensive qualitative overview of the hepatic vasculature. UFUS also facilitates quantitative measurements of the microvasculature, aiming to enhance our understanding of vascular disease mechanisms.

Non-invasive in vivo imaging of the vasculature is a powerful tool for studying disease mechanisms in rodents. To achieve high sensitivity imaging of the ...

Için Polikistik Böbrek Hastalığında Küçük Kemirgen modelleri Ultra yüksek Saha MRI Kullanımı

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