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).

قبل البدء في أي إجراءات مع الحيوانات الحية، يجب أن تتم الموافقة على البروتوكولات التجريبية من قبل لجنة رعاية الحيوان واستخدام المؤسسية (IACUC). 1. تكوين الماسح الضوئي <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> قبل البدء، تأكد من أن سخان هو في موقف OFF. </li><li style=";text-align:right;direction:rtl"> حدد التدرج التصوير صغير و38 ملم لفائف RF وحامل التصوير المصغر. </li><li style=";text-align:right;direction:rtl"> في تجويف المركزي للحامل تثبيت التجميع درجة حرارة متغيرة. </li></ol> 2. إعداد الحيوان <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> للتجارب التصوير بالرنين المغناطيسي، وتحقيق التخدير الأمثل باستخدام الأيزوفلورين المتبخرة. لتحريض التخدير، ووضع الحيوان في غرفة الاستقراء واصطف مع الأنسجة الماصة. ضبط مقياس الجريان من المرذاذ الأيزوفلورين إلى 2،0-2،5 لتر / دقيقة، والأيزوفلورين إلى 3٪ في الأكسجين. </li><li style=";text-align:right;direction:rtl"> إزالة أي علامة معدنية أو جسم معدني آخر في هذه المرحلة. تطبيق البيطري مرهم على عيون الحيوان لمنع جفاف بينما تحت التخدير. </li><li style=";text-align:right;direction:rtl"> مرة واحدة الحيوانوصلت الطائرة من التخدير الجراحي (أي فقدان الانسحاب المنعكس إلى أخمص قدميه قرصة)، ووضع الحيوان على حامل مع أنفها إدراجه ضمن مخروط الأنف. ضبط تدفق الهواء التخدير في التحقيق إلى 2،0-2،5 مل / دقيقة وتركيز الأيزوفلورين إلى 1،5-2،0٪ في الأكسجين. وسيتم تسليم التخدير من خلال مخروط الأنف أثناء العملية. دوري ضبط تركيز الأيزوفلورين اعتمادا على عمر الحيوان والوزن للحفاظ على معدل التنفس من ~ 40 نبضة في الدقيقة. </li><li style=";text-align:right;direction:rtl"> استخدام أصحاب الحيوانات لتأمين الحيوانات في مكان ومنع الحركة أثناء التجربة MRI. تختلف نوع من صاحب الحيوان اعتمادا على المنطقة الجسم التي يتم مسحها ضوئيا. ملاحظة: يمكن جعل أصحاب مخصص من مختبر البلاستيك (البولي بروبلين، تفلون، البوليسترين، البولي) لاستيعاب تجربة محددة وتتناسب مع حجم الحيوان (من الماوس الجديد إلى 160 غ الفئران). </li><li style=";text-align:right;direction:rtl"> وضع ميزان حرارة المستقيم في الحيوان لمراقبة درجة حرارة الجسم الحيوان. خلال إكسبeriment، والحفاظ على الحيوانات في 35-37 درجة مئوية، وذلك باستخدام تيار من الهواء الدافئ. ضبط درجة حرارة الهواء (30-38 درجة مئوية) وتدفق (1،200-2،000 L / ساعة) على أساس درجة حرارة الجسم ردود فعل الحيوان. </li><li style=";text-align:right;direction:rtl"> إرفاق بالون التنفسي استشعار الضغط على البطن الحيوان لمراقبة معدل التنفس. </li><li style=";text-align:right;direction:rtl"> تأمين الحيوان في وسط لفائف RF ووضع بعناية لفائف RF مع الحيوان إلى التصوير بالرنين المغناطيسي الماسح الضوئي. </li></ol> 3. MRI التجربة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> لحن، وتتناسب مع لفائف RF قبل بدء التجارب للحد من قوة RF المستخدمة وتعظيم نسبة الإشارة إلى الضوضاء. لبدء مطابقة / ضبط: <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> فتح أداة التحكم مطياف بالنقر على أيقونة أدوات. </li><li style=";text-align:right;direction:rtl"> في أداة التحكم مطياف انقر اقتناء → تمايل. تفتح نافذة ACQ / ريكو عرض منحنى تمايل. </li><li style=";text-align:right;direction:rtl"> بدلا من ذلك ضبط ضبط ومطابقة المكثفات (باستخدام ضبط ومطابقة قضبان) في خطوات صغيرة حتى السلطة RF تنعكستم تصغير. الهدف هو أن نرى منحنى مع حد أدنى على المحور الرأسي المتمركزة صفر على المحور الأفقي. </li><li style=";text-align:right;direction:rtl"> عندما تم تحقيقه معايرة من لفائف بنجاح، اضغط على زر الإيقاف في إطار ACQ / ريكو. </li></ol></li><li style=";text-align:right;direction:rtl"> الحصول على صور الكشفية في طائرات المتعامدة الثلاثة لخلق صور المحوري، الاكليلية والسهمي. استخدام تسلسل الصور السريعة مثل بوابة البينية السريعة انخفاض زاوية اطلاق النار (IG-FLASH) للحصول على الصور الكشفية 12. استخدام الصور الكشفية لضبط هندسة المناسبة للتصوير الفعلي. </li><li style=";text-align:right;direction:rtl"> تبعا لأهداف بحثية محددة، حدد السليم تسلسل الصور والمعلمات وبدء المسح الضوئي مع ضوء حركة المرور. وهذا معايرة قناة RF، شيم المغناطيس، تعيين تردد الناقل على صدى للمياه وضبط كسب المتلقي، كل تلقائيا. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> للدراسات التشريحية والصور T2 المرجحة، والحصول على شريحة متعددة في 2D أو 3D واسطة. لاختصار الوقت تجربة لقرار المكاني معين، والحفاظ على الميدان-of الرؤية (فوف) صغيرة ممكن ولكن كبيرة بما يكفي لتجنب القطع الأثرية التفاف حول (2،56 حتي 3،2 سم). </li></ol></li><li style=";text-align:right;direction:rtl"> تستمر دائرة تسلسل اختيار أقصر قليلا من دورة تنفس الحيوان عن طريق الانتقاء السليم لوقت التكرار (TR) و / أو عدد شرائح. وهذا يضمن أن البيانات يتم جمعها خلال فترة هدوء الحيوانات. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> على سبيل المثال، للصور في البطن، والحفاظ على معدل التنفس الحيوان في ~ 30 نبضة في الدقيقة. هذا هو حوالي 2000 ميللي ثانية في التنفس. استخدام توربو سرعة اقتناء مع تعزيز الاسترخاء (نادر) تسلسل واكتساب 11-19 شرائح الاكليلية، مع TR / TE 1500/9 ميللي ثانية، عامل نادر 8 و (مصفوفة 256 × 256، FOV 2.56 X 2.56 سم، وسمك شريحة 0.75 مم) . ملاحظة: من خلال تعديل TR إلى 1500 ميللي ثانية، والحفاظ على معدل التنفس الحيوان ~ 30 نبضة في الدقيقة (2000 ميللي ثانية في التنفس)، ونحن تأكد من أن البيانات التي تم جمعها خلال فترة هدوء الحيوانات. </li></ol></li><li style=";text-align:right;direction:rtl"> بعد أن تم الانتهاء من جميع الحصول على الصور، ضع الحيوان الممسوحة ضوئياعلى لوحة ساخنة ورصد حتى الإسعافية. بعد الشفاء، والعودة الحيوان إلى القفص ورصد ما لا يقل عن 1 ساعة قبل أن تعود إلى منشأة الحيوان. </li></ol>

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The authors have nothing to disclose.

وتظهر هذه المخطوطة جدوى استخدام UHF MRI كأداة لفي الجسم الحي توصيف phenotypical أو مراقبة المخدرات في نماذج القوارض لPKD. وصفنا التجارب القيام به في 16.4 T مع تحمل واسعة سميت بال المقدمة III عالية الدقة NMR مطياف مجهزة مع الملحقات التصوير ال...

مرض الكلى المتعدد الكيسات (PKD) يتضمن مجموعة من الاضطرابات أحادية المنشأ تتميز تطوير الكيسات الكلوية. فيما بينها هي من جسمي قاهر مرض الكلى المتعدد الكيسات (ADPKD) وراثي متنحي تكيس أمراض الكلى (ARPKD)، والتي تمثل الأنواع الأكثر شيوعا 1،2. ADPKD، الشكل الأكثر شيوعا من أمراض الكلى الكيسي وراثية، ونشأت عن طفرات في PKD1 أو PKD2 الجينات. ويتميز هذا في وقت متأخر من البداية، متعددة الكيسات الكلوية الثنائية، يرافقه متغير الخراجات خارج الكلى، فضلا عن تشوهات الهيكل العظمي القلب والأوعية الدموية والعضلات. ARPKD، الأكثر شيوعا التي تؤثر على الأطفال حديثي الولادة والأطفال الصغار، والتي تسببها طفرات في PKHD1 ويتميز الكلى مولد للصدى تضخم وتليف كبدي خلقي 3. الأهم من ذلك، يتميز ADPKD من قبل التجانس، سواء في الجين (الجيني) وطفرة (أليلية) المستويات، مما يؤدي إلى ص كبيرتقلب henotypic. ترتبط طفرات في الجين PKD1 مع العرض شديد السريري (العديد من الخراجات، والتشخيص المبكر وارتفاع ضغط الدم، وبيلة ​​دموية)، فضلا عن التقدم السريع لإنهاء مرحلة المرض الكلوي (قبل 20 سنة من المرضى الذين يعانون من الطفرات PKD2) 4. مرض شديد تكيس الكبد (الملعوبة) وتشوهات الأوعية الدموية يمكن أن تترافق مع طفرات في كل من PKD1 وPKD2 5. غالبية المضاعفات الكلوية من ADPKD تنشأ أساسا نتيجة للتوسع الكيس جنبا إلى جنب مع التهاب وتليف المرتبطة بها. تبدأ تطوير الكيس داخل الرحم وتستمر خلال فترة حياة المريض. الكلى عادة الحفاظ على شكل كلوي الشكل على الرغم من أنهم يمكن أن تصل إلى أكثر من 20 أضعاف حجم الكلى العادي. معظم المرضى الحالي توزيع الثنائي من الكيسات الكلوية، ولكن في بعض الحالات غير عادية، والكيس قد تتطور في نمط أحادي الجانب أو غير المتماثلة. يشكل تحديا كبيراشركة جنرال الكتريك لالكلى عقب المرضى الذين يعانون من ADPKD أو تنفيذ العلاجات هو التاريخ الطبيعي للمرض. خلال معظم مجراه، تبقى وظيفة الكلى طبيعية وبحلول الوقت الذي تبدأ وظيفة الكلى في الانخفاض، فإن معظم الكلى قد حلت محلها الخراجات. عندما يتم تنفيذ العلاجات في مراحل لاحقة، فإنه من غير المحتمل أن تكون ناجحة منذ المريض قد بالفعل قد وصلت إلى نقطة اللاعودة في مرض الكلى المزمن. في المقابل، عندما بدأ العلاج في المراحل المبكرة، فإنه من الصعب تحديد استجابة تستند فقط على معدل الترشيح الكبيبي. ونتيجة لذلك، فإن فكرة حجم الكلى كعلامة من تطور المرض اكتسبت اهتمام. وقد أظهرت التجمع من أجل إشعاعي الدراسات التصوير من مرض الكلى المتعدد الكيسات الدراسة (CRISP) أنه في المرضى الذين يعانون من ADPKD الزيادة في الكلى والكيس كميات يرتبط بشكل مباشر مع الكلوي تدهور وظيفة، مع التشديد على أهمية إجمالى حجم التداول الكلى (TKV) كماعلامة urrogate لتطور المرض 6،7. ونتيجة لذلك، يتم استخدام TKV حاليا منصب نقطة النهاية الأولية أو الثانوية في تجارب سريرية متعددة لADPKD 2،8،9. ألقت نماذج الفئران متعددة بما في ذلك الطفرات العفوية وراثيا الضوء على التسبب في PKD 10،11. أصبحت PKD1 أو Pkd2 نماذج (الطفرات في أي PKD1 أو Pkd2) الأكثر شعبية، لأنها الأمراض التي تصيب البشر تقليد تماما. بالإضافة إلى ذلك، تم استخدام نماذج القوارض مع الطفرات في جينات أخرى من PKD1 أو Pkd2 الجينات باعتباره منصة تجريبية لتوضيح مسارات الإشارات المتعلقة بهذا المرض. بالإضافة إلى ذلك، تم استخدام العديد من هذه النماذج لاختبار العلاجات المحتملة. ومع ذلك، فإن العامل المحدد في العديد من الدراسات القوارض لPKD هو في كثير من الأحيان عدم وجود طرق غير الغازية فعالة لتحليل بالتتابع التغيرات التشريحية والوظيفية في الكلى. ص المغناطيسيالتصوير esonance (MRI) هو الأسلوب الحالية التصوير معيار الذهب لمتابعة المرضى ADPKD، وتوفير ممتازة النقيض من الأنسجة الرخوة والتفاصيل التشريحية، والسماح للقياسات TKV. على الرغم من أن التصوير بالرنين المغناطيسي هي راسخة للتصوير التشريحية في الحيوانات والبشر أكبر، القوارض الصغيرة التصوير في الجسم الحي تنطوي على تحديات تقنية إضافية، حيث القدرة على الحصول على صور عالية الدقة قد يحد من فائدته. مع إدخال الحقل فائق (UHF) MRI (7-16،4 T) وتطوير التدرجات أقوى، أصبح من الممكن الآن لتحقيق أعلى إشارة إلى نسبة الضوضاء والتحليل المكاني للصور MRI مع نوعية التشخيص مماثلة لتلك التي التي تم الحصول عليها في البشر. ونتيجة لذلك، أصبح استخدام UHF MRI للتصوير في الجسم الحي من نماذج القوارض الصغيرة لPKD أداة قوية للباحثين.

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			<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>
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			<td>TopSpin2.0PV&#160;</td>
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			<td>VTU BVT 3000 digital</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.

في هذه المخطوطة، ونحن نهدف لإظهار فائدة UHF MRI كأداة لفي الجسم الحي توصيف phenotypical أو مراقبة المخدرات في نماذج القوارض لPKD وأمراض الكلى الأخرى. وكانت جميع التجارب جزءا من البروتوكولات التجريبية التي وافق عليها IACUC. في الجسم الحي...

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.

استخدام فائقة MRI الميداني في القوارض الصغيرة نماذج من مرض الكلى المتعدد الكيسات ل<em&gt; في فيفو</em&gt; Phenotyping والدواء المراقبة

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 Views.  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

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.

و في الجسم الحي النموذجي القوارض من الإصابات الناجمة عن انكماش ، وغير الغازية ورصد الانتعاش

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

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.

الطيفية Karyotyping إلى دراسة حالة شذوذ الكروموسومات في البشر والفئران مع مرض الكلى المتعدد الكيسات

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.

استخدام الخلايا الليفية الإنسان الأساسية لرصد نمط ظاهري الميتوكوندريا في مجال مرض باركنسون

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.

تقييم وظيفة القلب والصرف عضلة القلب عن طريق الحيوانات الصغيرة نظرة خزانة الاسترداد عكس (صلي) التصوير بالرنين المغناطيسي في الجرذان

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.

في الجسم الحي 19 F التصوير بالرنين المغناطيسي لتتبع الخليوي

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.

نموذج الزرد ممكن من مرض الكلى المتعدد الكيسات: ضربة قاضية لل<em&gt; wnt5a</em&gt; أسباب الخراجات في الكلى اسماك الزرد

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.

فيفو السابقين المعوية الحويصلات لتقييم الأغشية المخاطية النفاذية في نماذج من الأمراض المعوية

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.

3D الكمي<em&gt; في السيليكو</em&gt; النمذجة (q3DISM) من الدماغيه اميلويد بيتا البلعمة في نماذج القوارض من مرض الزهايمر

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.

في فيفو تقييم التنمية دشبذ الكسر أثناء التئام العظام في الفئران باستخدام جهاز الصدمة المتوافقة مع التصوير بالرنين المغناطيسي لعظم الفخذ الماوس

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