Name: a mouse 5/6th nephrectomy model that induces experimental uremic cardiomyopathy
Uploaded: 2017-11-07T12:30:00
Description: Watch this Scientific Journal Video about A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy at JoVE.com

Diese Arbeit wurde von NIH R15 1R15DK106666-01A1 (, J. Liu) und NIH RO1 HL071556 (, J.I Shapiro) unterstützt.

alle Tierpflege und Experimente wurden von der Marshall University institutionelle Pflege der Tiere und Nutzung Committee (IACUC) gemäß dem National Institute of Health (NIH)-Leitfaden für die Pflege und Verwendung von Labortieren genehmigt. Männliche C57BL/6 Mäusen (10-12 Wochen alt) untergebracht waren, in ein Erreger frei Tierstation in bestimmten Zimmern ausgestattet mit Käfigen, die gereinigte Luft unter einer 12 h hell/dunkel-Zyklus zu liefern. Nahrung und Wasser waren mitgelieferten Ad Libitum....

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S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+the+epidemic+of+cardiovascular+disease+in+chronic+renal+disease:+what+do+we+know?+What+do+we+need+to+learn?+Where+do+we+go+from+here?+National+Kidney+Foundation+Task+Force+on+Cardiovascular+Disease.">Controlling the epidemic of cardiovascular disease in chronic renal disease: what do we know? What do we need to learn? Where do we go from here? National Kidney Foundation Task Force on Cardiovascular Disease.</a> Am J Kidney Dis. 32 (5), 853-906 (1998).</li><li>Schiffrin, E. L., Lipman, M. L., Mann, J. F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+Kidney+Disease.">Chronic Kidney Disease.</a> Effects on the Cardiovascular System. 116 (1), 85-97 (2007).</li><li>Foley, R. 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M., Himmelfarb, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tipping+the+redox+balance+of+oxidative+stress+in+fibrogenic+pathways+in+chronic+kidney+disease.">Tipping the redox balance of oxidative stress in fibrogenic pathways in chronic kidney disease.</a> Pediatr Nephrol. 24 (12), 2309-2319 (2009).</li><li>Himmelfarb, J., Stenvinkel, P., Ikizler, T. A., Hakim, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+elephant+in+uremia:+oxidant+stress+as+a+unifying+concept+of+cardiovascular+disease+in+uremia.">The elephant in uremia: oxidant stress as a unifying concept of cardiovascular disease in uremia.</a> Kidney Int. 62 (5), 1524-1538 (2002).</li><li>Becker, B. N., Himmelfarb, J., Henrich, W. L., Hakim, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reassessing+the+cardiac+risk+profile+in+chronic+hemodialysis+patients:+a+hypothesis+on+the+role+of+oxidant+stress+and+other+non-traditional+cardiac+risk+factors.">Reassessing the cardiac risk profile in chronic hemodialysis patients: a hypothesis on the role of oxidant stress and other non-traditional cardiac risk factors.</a> J Am Soc Nephrol. 8 (3), 475-486 (1997).</li><li>Drummond, C. 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T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monoclonal+antibody+against+marinobufagenin+reverses+cardiac+fibrosis+in+rats+with+chronic+renal+failure.">Monoclonal antibody against marinobufagenin reverses cardiac fibrosis in rats with chronic renal failure.</a> Am J Hypertens. 25 (6), 690-696 (2012).</li><li>Tian, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spironolactone+attenuates+experimental+uremic+cardiomyopathy+by+antagonizing+marinobufagenin.">Spironolactone attenuates experimental uremic cardiomyopathy by antagonizing marinobufagenin.</a> Hypertension. 54 (6), 1313-1320 (2009).</li><li>Liu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuation+of+Na/K-ATPase+Mediated+Oxidant+Amplification+with+pNaKtide+Ameliorates+Experimental+Uremic+Cardiomyopathy.">Attenuation of Na/K-ATPase Mediated Oxidant Amplification with pNaKtide Ameliorates Experimental Uremic Cardiomyopathy.</a> Sci Rep. 6, 34592 (2016).</li><li>Yan, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+reactive+oxygen+species+in+a+feed-forward+mechanism+of+Na/K-ATPase-mediated+signaling+transduction.">Involvement of reactive oxygen species in a feed-forward mechanism of Na/K-ATPase-mediated signaling transduction.</a> J Biol Chem. 288 (47), 34249-34258 (2013).</li><li>Leelahavanichkul, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiotensin+II+overcomes+strain-dependent+resistance+of+rapid+CKD+progression+in+a+new+remnant+kidney+mouse+model.">Angiotensin II overcomes strain-dependent resistance of rapid CKD progression in a new remnant kidney mouse model.</a> Kidney Int. 78 (11), 1136-1153 (2010).</li><li>Ma, L. J., Fogo, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Model+of+robust+induction+of+glomerulosclerosis+in+mice:+importance+of+genetic+background.">Model of robust induction of glomerulosclerosis in mice: importance of genetic background.</a> Kidney Int. 64 (1), 350-355 (2003).</li></ol>

Die Ratte 5/6th PNx Modell ist weit verbreitet, CKD zu studieren. Aufgrund der viel kleineren Niere Größe in der Maus sind die klassischen Arterie Ligatur und Pol Resektion in Mausmodellen mit möglichen hohen Sterblichkeitsraten und unerwartete Blutungen/Blutverlust sehr anspruchsvoll.
Wir beschlossen ein Mausmodell PNx mit Pol Ligatur der Blutungen/Blutverlust zu überwinden. Dieses PNx-Modell nimmt weniger Zeit mit verbesserten Überlebensrate und hohe Reproduzierbarkeit. Diese...

CKD, auch bekannt als chronische Niereninsuffizienz ist eine fortschreitende Verlust der Nierenfunktion im Laufe der Zeit, die schließlich zur ständigen Nierenversagen entwickelt. CKD, vom frühen Stadium Nierenkrankheit Staaten terminaler Niereninsuffizienz (ESRD), ist ein großer Risikofaktor für Herz-Kreislauf-Krankheit und Sterblichkeit, und entwickelt sich zunehmend zu der klinischen Phänotyp genannt "urämischen Kardiomyopathie"1, 2,3. Die urämische Kardiomyopathie bei Patienten mit CKD oder terminaler Niereninsuffizienz ist verbunden mit Herz-Kreislauf-Anomalien, hauptsächlich verursacht durch Überlastung des linken Ventrikeldruck (LV) und/oder Volumen, was zu LV-Hypertrophie (LVH), LV-Dilatation und LV systolische Dysfunktion4 ,5,6. Kardialer Fibrose ist eine weitere gemeinsame pathologischen Prozess der urämischen Kardiomyopathie, die kardialen Compliance LV diastolische Dysfunktion wiederum reduziert. Schwere kardiale Fibrose führt zu plötzlichen Herztod sogar in diesen ohne kardiale Symptome7.
5/6th PNx ist eine häufig verwendete CKD-Tiermodell für Tierversuche mit Nierenversagen, urämischen Kardiomyopathie und Bluthochdruck. PNx wird durch Ablation von 5/6 renale Parenchym erreicht. Die Rattenmodell entwickelte sich zunächst mit den beiden am häufigsten Protokolle eingesetzt, chirurgische Resektion oder Infarkt. PNx Rattenmodell ist ein äußerst nützliches Modell urämischen Kardiomyopathie mit erheblichen Erhöhungen des Blutdrucks, Herzhypertrophie und Beeinträchtigung der diastolischen Funktion zu studieren. Später wurden PNx Mausmodelle, betrieben mit ähnlichen Techniken wie die Rattenmodell, aufgrund der breiten Verfügbarkeit und Benutzerfreundlichkeit machen genetische Manipulationen im Maus-System entwickelt.
Es ist gut dokumentiert, dass systemische oxidativen Stress ein konstantes Merkmal der beiden klinischen und experimentellen urämischen Kardiomyopathie8,9 ist. Des weiteren oxidativen Stress trägt zur urämischen Syndrom10und spielt eine entscheidende Rolle in der Pathogenese der kardialen Anomalien gesehen im urämischen Kardiomyopathie11,12,13. Zu diesem Punkt haben wir gezeigt, dass das Nagetier 5/6th PNx Modell bewirkt, dass physiologische, morphologische und biochemische Funktionen von urämischen Kardiomyopathie14,15,16, 17. in dem PNx Mausmodell hier beschriebenen, PNx betrieben Mäuse entwickelten bedeutenderen oxidativen Stress, zumindest teilweise vermittelte von Na/K-ATPase Signalisierung-Funktion, die in PNx-vermittelten experimentelle urämischen Kardiomyopathie ankommt. Dämpfung der Na/K-ATPase Signalisierung nicht nur reduziert oxidativen Verstärkung, sondern auch mildert die phänotypischen Veränderungen in PNx-vermittelten experimentelle urämischen Kardiomyopathie18.

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			<td>www.pilgtimmedical.com</td>
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			<td>Hilltop Lab Animals Inc.</td>
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			<td>Vevo 1100</td>
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			<td>FUJIFILM VisualSonics Inc.</td>
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			<td>Crystal Chem. Inc.</td>
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			<td height="21" style="height:21px;">Mouse BUN ELISA kit&#160;</td>
			<td>MyBioSource Inc&#160;</td>
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a mouse 5/6th nephrectomy model that induces experimental uremic cardiomyopathy

Chronische Niereninsuffizienz (CNI) ist ein großer Risikofaktor für Herz-Kreislauf-Krankheit und Sterblichkeit und entwickelt sich zunehmend zu der klinischen Phänotyp "urämischen Kardiomyopathie" genannt. Wir beschreiben hier ein experimentelle CKD Maus-Modell namens 5/6th partiellen Nephrektomie (PNx) mit Pol Ligatur, die urämischen Kardiomyopathie auf vier Wochen nach der Operation entwickelt. Dieses PNx-Modell wurde von einer Operation in zwei Schritten durchgeführt. In Schritt eins Chirurgie waren beide Pole der linken Niere ligiert. In Schritt zwei Operationen, die 7 Tage nach der Schritt-eine Operation durchgeführt wurde, wurde die rechte Niere entfernt. Für die Schein-Chirurgie die gleichen Eingriffe erfolgten aber ohne Pol Ligatur der linken Niere oder Entfernung von der rechten Niere. Chirurgische Eingriffe sind einfacher und zeitsparender, im Vergleich zu anderen Methoden. Aber der Rest funktionellen Niereninsuffizienz Masse ist nicht so leicht als der Nierenarterie Ligatur gesteuert. Vier Wochen nach der Operation im Vergleich zu den Schein betrieben Mäusen die PNx Mäuse entwickelt, eingeschränkter Nierenfunktion, Anämie, Herzhypertrophie, kardialer Fibrose und sank der systolische und der diastolische Herzfunktion.

Die Daten zeigten, dass dies geändert 5/6th PNx durch Pol Ligatur ist ein einfaches und effektives Modell urämischen Kardiomyopathie zu untersuchen. Vier Wochen nach der Operation dieses Model präsentiert PNx beeinträchtigte Nierenfunktion, Anämie, Herzhypertrophie, kardialer Fibrose und sank der systolische und der diastolische Herzfunktion. Die Ergebnisse sind nachfolgend zusammengefasst.
Vier ...

Watch this Scientific Journal Video about A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy at JoVE.com

A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy

Diese Handschrift enthält eine detaillierte zweistufigen chirurgische Prozedur zum Maus 5/6th partiellen Nephrektomie (PNx) mit Pol Ligatur durchzuführen. Vier Wochen nach der Operation im Vergleich zu Schein betrieben Mäuse, die PNx Mäuse entwickelt, eingeschränkter Nierenfunktion, Anämie, Herzhypertrophie, kardialer Fibrose und sank der systolische und der diastolische Herzfunktion.

5/6th Nephrektomie Mausmodell, die experimentelle urämischen Kardiomyopathie induziert

Chronic kidney disease (CKD) is a great risk factor for cardiovascular disease events and mortality, and progressively develops to the clinical phenotype ...

The overall goal of this procedure is to set up a simple mouse five-sixth partial nephrectomy model with pole ligation with a better survival rate than traditional models. This method can help answer key questions in the chronic kidney disease field about the development and potential therapeutic treatment of uremic cardiomyopathy. The main advantage of this technique is that it is an easy and consistently reproducible animal model that mimics the pathology of chronic kidney disease in human patients.Generally, individuals new to this method will struggle because of the much smaller kidneys, blood vessels, and ureters in the mouse. Begin by confirming the appropriate level of sedation by a lack of response to toe pinch and applying lubricant to the mouse's eyes. Place the mouse on a warming pad and insert a rectal thermometer to maintain the body temperature around 37 degrees Celsius during the procedure.Then, place the animal onto its right side and shave the hair over the surgical area. Remove any remaining hair with depillatory cream and disinfect the exposed skin with polyhydroxy dine solution and 70%ethanol. Align an autoclave surgical linen over the surgery site.And make an approximately one-centimeter lumbar incision through the skin and muscle tissue. Using the scalpel and forceps, separate the muscle and fascia to expose the left kidney. Then, use small blunt forceps to gently pull on the perirenal fat to extract the kidney from the abdominal cavity.Gently tease the connective tissue and adrenal gland from the superior end of the kidney towards the middle pylorus of the organ where the blood vessels enter and exit the renal tissue. Isolate the ureter around the inferior half of the kidney within the connective tissue, and gently separate the connective tissue from the kidney to ensure that the ureter is not ligated. It is essential to separate the connective tissue carefully so that the ureter is not damaged and does not bleed.Isolate the ureter around the inferior half of the kidney within the connective tissue. Then, use three zero silk suture strings to ligate the kidney approximately 0.4 centimeters from the superior portion, toward the middle of the pylorus and 0.4 centimeters from the inferior portion towards the pylorus. When tightening the sutures, apply just enough tension to compromise the blood supply to the distal portion of the kidney without causing bleeding on either side.If no bleeding is observed after two minutes, gently return the kidney to its original anatomic location, and use four zero and three zero sutures to close the muscle and skin layers respectively. Then, apply antibiotic ointment to the surgical area and house the mouse individually with a heat lamp and monitoring until sternal recumbency. Seven days after the polar ligation, expose the right kidney as just demonstrated.Use blunt forceps to clear the surrounding fat and connective tissue, and loosely place two three zero silk sutures around the renal artery and vein. Gently tease the connective tissue and adrenal gland from the superior end of the kidney towards the middle pylorus. Gently displacing the ureter from the inferior end of the tissue, untie all of the vessels together without causing bleeding.Next, move one loose ligature knot toward the abdominal aorta side, and the second loose ligature toward the kidney side. And tie off the abdominal aorta ligature with double knots. A solid knot will change the color of the right kidney.Tie off the second ligature knot with double knots, leaving spaces between the knots so the vessels can be cut, and cut the renal vessels. After removing the kidney, close the muscle and skin layers as just demonstrated. Then, dry the kidney with sterile gauze and weigh the harvested renal tissue.At four weeks post-surgery, the partial nephrectomy mice demonstrate significantly increased levels of cyctatin C, creatinine, and blood urea nitrogen in the plasma compared to sham surgery animals. Partial nephrectomy stimulated anemia is also apparent, as evidenced by the significantly lower hematocrit levels in the polar ligated animals. Cardiac hypertrophy and diastolic dysfunction in the partially nephrectomized mice are indicated by an increased heart-to-body-weight ratio and by transthoracic echocardiography analysis which reveals significant increases in the posterior and anterior wall thicknesses, the myocardial performance index, and the left ventricle mass index.Partial nephrectomy mediated cardiac fibrosis is also demonstrated by increases in type 1 collagen expression in left ventricle homogenates and left remnant kidney homogenates. Further, oxidant stress places an important role in the experimental uremic cardiomyopathy, significantly stimulating direct protein carbonylation modification of multiple proteins and proto-oncogene tyrosine-protein kinase activation in the left ventricle homogenates. Once mastered, this technique can be completed in 40 minutes if it is performed properly.While attempting this procedure, it's important to remember to check the respiration rate of the animal and to adjust the isofluranes as appropriate. Following this procedure, other experiments like comparing the effects of specific drug treatments on partial nephrectomy recovery can be performed to answer traditional questions about specific therapeutic treatments of interest. After its development, this technique paved the way for researchers in the field of chronic kidney disease to explore the pathology of and potential drug screening in uremic cardiomyopathy.After watching this video, you should have a good understanding of how to set up a partial nephrectomy model in the mouse.

Research

JoVE Journal

Biology

A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy

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In-vivo-Imaging und therapeutische Behandlungen in einem orthotopen Mausmodell des Eierstockkrebs

Human cancer and response to therapy is better represented in orthotopic animal models.  This paper describes the development of an orthotopic mouse model ...

Forschung

Biologie

Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal

Oct4GiP Reporter Assay auf Gene, die Maus embryonalen Stammzellen Wartungs-und Self-Study Erneuerung Regulate

Pluripotency and self-renewal are two defining characteristics of embryonic stem cells (ES cells). Understanding the underlying molecular mechanism will ...

An Analytical Tool that Quantifies Cellular Morphology Changes from Three-dimensional Fluorescence Images

Ein analytisches Werkzeug, das zelluläre Morphologie Änderungen von Dreidimensionale Fluorescence Images Quantifiziert

The most common software analysis tools available for measuring fluorescence images are for two-dimensional (2D) data that rely on manual settings for ...

Strategies for Tracking Anastasis, A Cell Survival Phenomenon that Reverses Apoptosis

Strategien für die Spurhaltung Anastasis, einer Zelle Überleben Phänomen, das Apoptose Kehrt

Anastasis (Greek for &#8220;rising to life&#8221;) refers to the recovery of dying cells. Before these cells recover, they have passed through important ...

Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates

Trans-innere Zelle Masse Injektion von embryonalen Stammzellen führt zu höheren Chimerism Raten

In an effort to increase efficiency in the creation of genetically modified mice via ES Cell methodologies, we present an adaptation to the current ...

A Mouse Model of Intestinal Partial Obstruction

Ein Maus-Modell der teilweisen Darmverschluss

Intestinal obstructions, that impede or block peristaltic movement, can be caused by abdominal adhesions and most gastrointestinal (GI) diseases including ...

Purification of Platelets from Mouse Blood

Reinigung von Plateletten aus dem Mausblut

Platelets are purified from whole blood to study their functional properties, which should be free from red blood cells (RBC), white blood cells (WBC), ...

A Laser Capture Microdissection Protocol That Yields High Quality RNA from Fresh-frozen Mouse Bones

Ein Laser Capture Microdissection Protocol, das hochwertige RNA aus frisch gefrorenen Mausknochen liefert

RNA yield and integrity are decisive for RNA analysis. However, it is often technically challenging to maintain RNA integrity throughout the entire laser ...

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

Massenspektrometrieanalyse zur Identifizierung der Ubiquitylierung von EIFP-A mit EYFP-Tags (EYFP-CENP-A)

Studying the structure and the dynamics of kinetochores and centromeres is important in understanding chromosomal instability (CIN) and cancer ...

Skeletal Phenotype Analysis of a Conditional Stat3 Deletion Mouse Model

Skeletal Phenotype Analysis of a Conditional Stat3 Deletion Mouse Model

Skelett-Phänotyp-Analyse eines bedingten Stat3-Löschmausmodells

Transgenic mouse models are powerful for understanding the critical genes controlling osteoclast differentiation and activity, and for studying mechanisms ...