Name: a murine model of irreversible and reversible unilateral ureteric obstruction
Uploaded: 2014-12-20T15:00:00.000+00:00
Duration: 14 min 5 s
Description: Watch this Scientific Journal Video about A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction at JoVE.com

The present study was supported by grants from Kidney Research UK (ST4/2011), the Cunningham Trust (CT11/14), the Mrs AE Hogg Charitable Trust for Kidney Research and the Renal Endowment Fund of the Royal Infirmary of Edinburgh.

일반 지침 : 동물 실험은 동물에 의해 부과 된 지침과 규정에 따라 (과학적 절차) 법 1986이 프로토콜 많은에서 수행 할 수있는 동반 비디오 프로토콜 표준 UUO 및 R-UUO 모두이며, 수행되었다 마우스 균주. 첨부 된 비디오에서 두 절차는 남성 C57BL / 6 마​​우스 세 팔주에서 수행됩니다. 대표적인 결과 섹션에 제시된 데이터는 남성 FVB / N 마우스로부터 얻었다. 주 :이 프로토콜 및 표준 방법을 수행 UUO R-UUO 왼쪽 요관을 이용하여 비디오 정보를 수반하지만 동일한 기술을 쉽게 오른쪽 요관에 적용될 수있다. 1. 동물 준비 및 개복술 <ol><li> 살균 (멸균) 장비 및 소모품 모든 절차를 수행합니다. </li><li> 마취 복강 케타민 염산염 (70 ㎎ / kg) 및 medetomidine 염산염 (1 ㎎ / kg)을 주사마우스. 참고 : 결과 마취 평면의 지속 시간은 4 시간이며, 더 추가 마취가 필요하지 않습니다. </li><li> 반사 예를 들어, 발가락 핀치의 손실에 의한 마취의 깊이를 확인합니다. </li><li> 절개 주변 지역 모든 머리를 제거하고 희석 클로르헥시딘 솔루션의 응용 프로그램을 통해 수술을 위해 복부 피부를 준비합니다. </li><li> 앙와위에서 가열 수술 패드에 마우스를 놓고 낮은 압정 접착 테이프를 사용하여 패드에 사지를 고정합니다. </li><li> 절차 동안 가열 패드 수술의 결과로서 열 화상 흔적 마우스를 모니터링. 가능하면 비 전기 열원을 사용한다. </li><li> 부 프레 노르 핀 염산염 (내지 0.06 ㎎ / kg)의 피하 주사에 의해 진통을 관리하고 각막 건조를 방지하기 위해 눈 윤활제를 적용합니다. </li><li> 정중선 개복 및 복강에 접근하기 위해 조직 분리 가위를 사용하여 무 혈관의 백선 알바의 절개를. </li><li> 마우스를 올려 놓와 콜리 브리를 삽입절개에 트랙터. </li></ol> 2. 일방적 Ureteric의 방해 <ol><li> 멸균 면봉을 사용하여 복강의 오른쪽으로 장을 변위에 의해 왼쪽 요관을 노출하고 젖은 커튼로 커버. </li><li> 각도 집게 격리하고 왼쪽 요관을 들어 올려 사용. </li><li> ureteric 방해를 만들려면 아무 곳이나 방광 및 신장 골반 사이의 6 / O 검은 꼰 실크 봉합사로 두 번 왼쪽 요관을 결찰. 장기 실험을 위해, 모든 복부 수술에 대한 흡수성 봉합사를 사용합니다. 대안 적으로, 결찰 클립 요관에 적용될 수있다. </li><li> 방광 요관으로부터 격리하기 위해 두 요관 봉합 사이를 나눈다. </li><li> 조심스럽게 복강에 장을 교체합니다. </li><li> 4 절에 나와있는 단계를 수행 - 수술 후 회복 및 관리는 절개를 닫고 마취를 반전 할 수 있습니다. </li></ol> Reversibl 3. 준비전자 일방적 Ureteric의 방해 <ol><li> 수술 마우스를 준비하고 단계 위의 1.1-2.2에서 설명하는대로 왼쪽 요관을 격리 할 것. </li><li> 반전 될 수있는 ureteric 방해를 만들려면, 방광에 가까운 6 / O 검은 꼰 실크 봉합사로 두 번 왼쪽 요관을 결찰. 이는 요관 주위 장소에 부드러운 벽 플라스틱 튜브를 고정하는 데 사용되는만큼 상부 봉합사의 한쪽 끝을 남겨주세요. </li><li> 그것은 그것이 요관에인가 될 수 있도록 오픈 스플레이 될 수 있도록 부드러운 벽의 플라스틱 튜브의 길이 5mm의 길이 방향 슬릿을 확인. 1mm와 2mm의 외경 내경 상관 벽으로 부드러운 실리콘 플라스틱 튜브가 사용될 수있다. </li><li> 요관 주위에 부드럽게 부드러운 벽 플라스틱 튜브를 놓습니다. 이 요관 주위를 닫으면 긴 봉합 튜브의 슬릿의 중심에서 나온다 있는지 확인합니다. </li><li> 요관을 둘러싸는 부드러운 벽으로 둘러싸인 튜브 주위에 6 / O 검은 꼰 실크 봉합사의 길이를 놓고 넥타이한 번. 이제 길이 방향으로는 이전에 묶여 봉합 위에 놓여 있도록 튜브를 통해, 튜브의 슬릿의 중심에서 신흥, 봉합의 긴 끝을 배치합니다. </li><li> 튜브 및 요관 주위 장소에 긴 봉합 끝을 고정 두 번 튜브 주위에있는 봉합사를 묶어. 주 : 요관 및 연질 플라스틱 배관 벽에 유착 형성을 방지하기 위해 접착 환원 액 배관 주변에 적용될 수있다. </li><li> 조심스럽게 복강에 장을 교체합니다. </li></ol> 4. 수술 후 복구 및 관리 <ol><li> 5 / O 검은 꼰 실크 봉합사를 사용하여 블랭킷 스티치와 복막을 닫고 금속 피부 클립을 사용하여 피부에 근접. </li><li> 수술 후 감염의 위험을 최소화하기 위해, 복부 피부에 요오드 / 알코올 용액으로서 방부제 등을 적용한다. </li><li> 피하 atipamezole 염산염 (2 ㎎ / kg)와 부분적으로 반대 마취. </lI&gt; <li> 1 ml의 피하 주사에 의해 유체를 관리 식염수를 따뜻하게. </li><li> 이 의식을 회복 할 때까지 마우스를 모니터링합니다. </li><li> 마우스가 24 시간 동안 29 ° C로 유지 가열 상자에 복구 할 수 있습니다. 적신 음식은 유체 및 영양 섭취를 장려하기 위해 제공 될 수있다. </li><li> 장애물이 원하는 수준의, 일반적으로 7 일 유도하기 위해 복구하기 위해 마우스를 둡니다. </li><li> 장기적 회복 실험에 지속적인 진통제를 제공한다. 마우스를 7 일 이상을 위해 회수 될 경우, 피부 클립을 수술 후 7 일 제거. </li><li> 폐색 원하는 레벨이 유도되면, 하나 후술하는 바와 같이, 역방향 UUO, 경부 탈구에 의해 마우스를 안락사 조직 병리학 분석 신장을 모은다. </li></ol> 5 가역 일방적 Ureteric의 방해 <ol><li> 단계에 설명 된대로 R-UUO가 수술을 위해, R-UUO을위한 준비를 마쳤습니다 마우스를 준비 수행하려면 10.1-1.7. </li><li> 존재하는 경우, 피부 클립을 제거하고 분할 또는 복강에 접근하기 복막에 봉합사를 제거한다. 주 :이 요관 장기 실험 피부 클립 방해 되었다면 응용 다음 7 일 제거 된 것이다. </li><li> 마우스를 준비 단계 1.8-2.2에서 설명한 바와 같이 왼쪽 요관을 격리. </li><li> 각도 집게를 사용하여 형성 할 수있는 모든 육아 종성 조직에서 부드러운 벽 플라스틱 튜브를 무료. </li><li> 메스 차단 된 요관 주위의 플라스틱 튜브를 들고 봉합을 잘라 튜브를 제거합니다. </li><li> 왼쪽 신장에 수신증의 존재를 평가하여 성공적인 UUO을 확인, 신장은 창백 나타납니다. </li><li> 봉합 사이의 요관을 나눈다. </li><li> 멸균 거즈의 작은 조각에 부착 신장 수뇨관의 나머지 길이를 놓는다. 이것은 수뇨관에서 소모 시키게 뇨 침전물과 사균을 수집하는 데 사용할차 봉합 한 번 신우가 제거됩니다. </li><li> 위의 요관을 분할하지만, 근처, 봉합사와는 신장이 거즈에 배출 할 수 있습니다. 방광에 가장 가까운 그대로, 낮은 봉합을 남겨주세요. 이은 소변이 방광과 복강으로 누출 없다는 것을 확인하는 것입니다. </li><li> 요관과 신우가 배출되고 나면, 요관의 남은 길이의 끝에 긴 6 / O 검은 꼰 실크 봉합사를 적용합니다. 이것은 나중에 수행 방광 문합에 요관을 돕기 위해 사용됩니다. </li><li> 방광 요관의 나머지 길이를 문합 아래 단계에 따라 : <ol><li> 그것의 원래 위치로 전방을 거짓말과 신장을 통해 거짓말을하도록 요관 돌립니다. </li><li> 그것이 방광의 방향으로 나온다되도록 요관의 단부로부터 하나의 8 / O 폴리 아미드 모노 필라멘트 봉합사를 점착 2mm를 놓는다. 봉합사는 근육 코트에 남아 있고 요관 루멘를 입력하지 않도록주의하십시오. 시침 봉합 줘야L은 방광 요관을 고정하기 위해 사용될 수. </li><li> 방광을 통해 채널을 생성하기 위해서는 방광의 정면 (복부) 벽쪽으로 종료하도록 방광을 통해 대각선으로 21G 바늘을 전달합니다. </li><li> 21G 바늘의 경사에 눈 바늘을 휴식. 방광을 통해 눈의 바늘을 안내하는 21G 바늘을 사용합니다. 눈 바늘은 방광을 통해 요관을 위해 사용됩니다. </li><li> 9 / O 시침 봉합사를 통과, 방광에서 밖으로 엔트리 포인트에 인접한 방광 벽을 통해 첫 절개를 통해, 단계 5.11.2에서 요관에 적용. 방광 요관이 통과되면 이는 방광 요관 앵커에 연결된다. </li><li> 눈 바늘을 통해 단계 5.10에서 요관의 끝 부분에 적용되는 긴 6 / O 검은 꼰 실크 봉합사를 놓습니다. </li><li> 조심스럽게 요관도 방광을 당겨을 보장하는 동안 방광 밖으로 눈 바늘을 철회. </li><li> 요관은 일에서 나온다되면전자 블래 눈동자 바늘을 제거하고 다시 방광 수축을 방지 요관의 끝에 긴 봉합 클램프를 적용. </li><li> 방광의 장소에서 요관을 고정하려면 단계 5.11.5에 적용되는 9 / O 폴리 아미드 모노 필라멘트 압정 봉합을, 넥타이. </li><li> 단단히 방광 내 요관을 고정하기 위해 진입 점을 중심으로 두 위치에서 단계 5.11.2 및 5.11.5에 설명 된대로 추가 한 9 / O 폴리 아미드 모노 필라멘트 타이 봉합을 적용합니다. </li><li> 열이 위의 뒷면을 사용하여 응급 요관의 더 많은 노출을 뒤로 살짝 방광 벽에 ​​밀어 넣습니다. </li><li> 요관의 끝에있는 긴 봉합 위의 요관을 나눈다. 요관 방광의 몸으로 철회해야한다. 이 요관의 오픈 루멘을 확인, 방광의 구멍에서 흐르는 소변을 관찰하는 것이 일반적이다. </li><li> 9 / O 폴리 아미드 모노 필라멘트 봉합사 방광에 상처 출구를 닫습니다. </li></ol></li><li> 조심스럽게에 장을 교체복강. </li><li> 단계 4.1-4.8에서 설명하는대로 수술 후 치료를 제공합니다. </li><li> 신장이 칠일 일반적으로 압축이 해제 될 때까지 마우스를 복구 할 수 있습니다. </li><li> 실험 종료 시점에서, 경부 탈구에 의해 마우스를 안락사 및 조직 병리학 적 분석을 위해 신장을 모은다. </li></ol>

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C., 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=Postobstructive+regeneration+of+kidney+is+derailed+when+surge+in+renal+stem+cells+during+course+of+unilateral+ureteral+obstruction+is+halted.">Postobstructive regeneration of kidney is derailed when surge in renal stem cells during course of unilateral ureteral obstruction is halted.</a> Am J Physiol Renal Physiol. 298, F357-F364 (2010).</li><li>Manson, S. R., Niederhoff, R. A., Hruska, K. A., Austin, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endogenous+BMP-7+is+a+critical+molecular+determinant+of+the+reversibility+of+obstruction-induced+renal+injuries.">Endogenous BMP-7 is a critical molecular determinant of the reversibility of obstruction-induced renal injuries.</a> Am J Physiol Renal Physiol. 301, F1293-F1302 (2011).</li><li>Manson, S. R., Niederhoff, R. A., Hruska, K. A., Austin, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+BMP-7-Smad1/5/8+pathway+promotes+kidney+repair+after+obstruction+induced+renal+injury.">The BMP-7-Smad1/5/8 pathway promotes kidney repair after obstruction induced renal injury.</a> J Urol. 185, 2523-2530 (2011).</li><li>Chaves, L. D., 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=Contrasting+effects+of+systemic+monocyte/macrophage+and+CD4++T+cell+depletion+in+a+reversible+ureteral+obstruction+mouse+model+of+chronic+kidney+disease.">Contrasting effects of systemic monocyte/macrophage and CD4+ T cell depletion in a reversible ureteral obstruction mouse model of chronic kidney disease.</a> Clin Dev Immunol. 2013, 836-989 (2013).</li><li>Haque, M. E., 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=Longitudinal+changes+in+MRI+markers+in+a+reversible+unilateral+ureteral+obstruction+mouse+model:+preliminary+experience.">Longitudinal changes in MRI markers in a reversible unilateral ureteral obstruction mouse model: preliminary experience.</a> J Magn Reson Imaging. 39, 835-841 (2014).</li><li>Bai, Z. M., 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=Arterially+transplanted+mesenchymal+stem+cells+in+a+mouse+reversible+unilateral+ureteral+obstruction+model:+in+vivo+bioluminescence+imaging+and+effects+on+renal+fibrosis.">Arterially transplanted mesenchymal stem cells in a mouse reversible unilateral ureteral obstruction model: in vivo bioluminescence imaging and effects on renal fibrosis.</a> Chinese Med J. 126, 1890-1894 (2013).</li><li>Chaabane, W., 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=Renal+functional+decline+and+glomerulotubular+injury+are+arrested+but+not+restored+by+release+of+unilateral+ureteral+obstruction+(UUO).">Renal functional decline and glomerulotubular injury are arrested but not restored by release of unilateral ureteral obstruction (UUO).</a> Am J Physiol Renal Physiol. 304, F432-F439 (2013).</li><li>Thornhill, B. A., Burt, L. E., Chen, C., Forbes, M. S., Chevalier, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variable+chronic+partial+ureteral+obstruction+in+the+neonatal+rat:+a+new+model+of+ureteropelvic+junction+obstruction.">Variable chronic partial ureteral obstruction in the neonatal rat: a new model of ureteropelvic junction obstruction.</a> Kidney Int. 67, 42-52 (2005).</li><li>Thornhill, B. A., Chevalier, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variable+partial+unilateral+ureteral+obstruction+and+its+release+in+the+neonatal+and+adult+mouse.">Variable partial unilateral ureteral obstruction and its release in the neonatal and adult mouse.</a> Methods Mol Biol. 886, 381-392 (2012).</li></ol>

저자는 더 경쟁 또는 공개 할 이해 관계 충돌이 없습니다.

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. The R-UUO model is of relevance to human disease as patients often present with obstruction of the renal tract secondary to prostatic hypertrophy, posterior urethral valves etc. that requires decompression with patients commonly left with significant residual kidney impairment that has no current treatment options and may eventually lead to end stage kidney disease.8,9 The models detailed here enable ...

The overall aim of the experimental model described here is to induce obstructive nephropathy by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). A simple irreversible model of UUO is presented in which the left ureter is permanently obstructed by ligation with a suture or by the application of a ligating clip. This results in marked dilatation of the ureter together with reduced renal blood flow and glomerular filtration. Renal histology demonstrates tubular dilatation and increasingly severe interstitial renal inflammation and fibrosis. Irreversible UUO is a useful model and has been adopted by many researchers in the study of both renal inflammation and fibrosis.1-4 Although the irreversible model of UUO requires some surgical expertise it is relatively straightforward and is often used to seek insights into the pathogenesis of interstitial renal injury and the ensuing fibrosis. Also presented is a less frequently used R-UUO model using a modification of the method originally described by Tapmeier et al.5 The R-UUO model has much future potential for the study of inflammatory and immune processes, cellular and tissue regeneration as well as the subsequent tissue remodeling following the removal of an injurious stimulus. 
The more challenging R-UUO model has been used by a limited number of investigators with some groups employing a significantly different surgical technique to that described here6,7 though with interesting results. In the R-UUO model presented, the ureter is ligated to induce complete ureteric obstruction for a period of time sufficient to induce the level of injury and fibrosis desired: 7 days of UUO was chosen in the method described here. The ureteric obstruction is reversed and the kidney is allowed to decompress for a period of time determined by the investigator before the mice are culled and the kidney removed for analysis: 7 days of decompression was chosen in the method described here though a longer period would be chosen if the resolution of inflammation and fibrosis was being studied. Although the method described here requires significant surgical expertise, it offers several advantages over other R-UUO models. The application of soft walled plastic tubing to the obstructed ureter prevents excessive ureteric dilation and this facilitates the subsequent manipulation and anastomosis of the ureter. Furthermore, in the described R-UUO model the ureter is divided thereby allowing the removal of any residual urinary sediment and debris. This confirms that the remaining ureter lumen is de-obstructed and patent prior to anastomosis to the bladder. 
Experiments incorporating both the irreversible and reversible UUO models can provide researchers with a powerful insight into the molecular and cellular mechanisms of both injury and subsequent resolution and regeneration. Thus, the model of R-UUO described here would be highly relevant to researchers interested in post-inflammatory tissue remodeling and how this can be modulated.

<table><tbody><tr><td>Blunt Dissecting Scissors</td><td>Fine Science Tools</td><td>14072-10</td><td></td></tr><tr><td>Spring Scissors &amp;ndash; straight</td><td>Fine Science Tools</td><td>15000-10</td><td></td></tr><tr><td>Toothed forceps</td><td>Fine Science Tools</td><td>11021-12</td><td></td></tr><tr><td>Angled forceps x 2</td><td>Fine Science Tools</td><td>00649-11</td><td></td></tr><tr><td>Straight forceps</td><td>Fine Science Tools</td><td>00632-11</td><td></td></tr><tr><td>Colibri 3 cm wire retractor</td><td>Fine Science Tools</td><td>17000-03</td><td></td></tr><tr><td>Castroviejo needle holder with lock</td><td>Fine Science Tools</td><td>12565-14</td><td></td></tr><tr><td>Wound clip applicator</td><td>Fine Science Tools</td><td>12031-07</td><td></td></tr><tr><td>7 mm wound clips</td><td>Fine Science Tools</td><td>12032-07</td><td></td></tr><tr><td>Castroviejo micro needle holder with lock</td><td>Fine Science Tools</td><td>12060-01</td><td></td></tr><tr><td>OPMI pico microscope</td><td>Carl Zeiss</td><td>S100</td><td></td></tr><tr><td>Heat electronic pad</td><td>Cozee Comfort</td><td>n/a</td><td></td></tr><tr><td>6/O silk braided suture</td><td>Harvard Apparatus</td><td>72-3287</td><td></td></tr><tr><td>9/O Dafilon (polyamide) suture</td><td>B-Braun</td><td>G1111434</td><td></td></tr><tr><td>5/O braided silk suture</td><td>Harvard Apparatus</td><td>51-7680</td><td></td></tr><tr><td>Regular bevel needle, 1 inch, 21G</td><td>Becton, Dickinson and Company</td><td>305175</td><td></td></tr><tr><td>1 ml syringe slip tip</td><td>Becton, Dickinson and Company</td><td>300184</td><td></td></tr><tr><td>Wypall paper swabs</td><td>Kimberley-Clark</td><td>L40</td><td>Sterilised (Autoclave)</td></tr><tr><td>Cotton wool buds</td><td>Johnson and Johnson</td><td>n/a</td><td>Sterilised (Autoclave)</td></tr><tr><td>Plain drapes</td><td>Guardian</td><td>CB03</td><td>Sterilised (Autoclave)</td></tr><tr><td>Soft wall silicone rubber tubing</td><td></td><td></td><td>Silicone tubing - internal diameter 1.0 mm; external diameter 2.0 mm; wall thickness 0.6 mm</td></tr><tr><td>(Lacri-Lube) White soft paraffin 57.3%, mineral oil 42.5% and lanolin alcohols 0.2%</td><td>Allergan Ltd</td><td>21956GB10X</td><td></td></tr><tr><td>(Videne) Povidone-iodine 10%</td><td>Ecolab Ltd</td><td>PL 04509/0041</td><td></td></tr><tr><td>(Vetalar V) Ketamine hydrochloride</td><td>Pfizer Animal Health</td><td>Vm 42058/4165</td><td>100mg/ml solution</td></tr><tr><td>(Domitor) Medetomidine hydrochloride</td><td>Orion Pharma</td><td>Vm 06043/4003</td><td>1mg/ml</td></tr><tr><td>(Vetergesic) Bupernorphine hydrochloride</td><td>Alsto Animal Health</td><td>Vm 00063/4002</td><td>0.3mg/ml</td></tr><tr><td>(Antisedan) Atipamezole hydrochoride</td><td>Orion Pharma</td><td>Vm 06043/4004</td><td>5mg/ml</td></tr><tr><td>(Adept) 4% Icodextrin</td><td>Baxter</td><td></td><td>Adhesion reduction solution</td></tr><tr><td>NaCl 0.9%</td><td>Baxter</td><td>FKE1323</td><td></td></tr></tbody></table>

a murine model of irreversible and reversible unilateral ureteric obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

신장의 모양이 변경 현저 ureteric 폐쇄를 다음과는 창백한와 시간 촉진에 시제 (그림 1)가된다. 장애물과 신우 요관의 근위부의 증가 팽창이있다. 신장은 신장 피질과 수질의 현저한 얇아 결과 연장 방해 폐쇄성 신증 증가의 지속 시간으로 점점 위축된다. ureteric 폐색의 반전에 따라, 신장의 색 혈류 증가 및 조직 부종 해소 할 어두워진다. 그들은 연장 팽만의 기간 (그림 1)의 결과로 약?...

Watch this Scientific Journal Video about A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction at JoVE.com

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

돌이킬 수 및 가역 일방적 Ureteric 폐색의 쥐 모델

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

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30.4K 조회수.  University of Edinburgh. The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

비디오: A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Creation of Reversible Cholestatic Rat Model

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and affect the nutritional status and surgical outcomes. Study of the pathophysiological changes occurring in the liver and other organs is of importance. Various studies have been done in cholestatic rat models.
We used a reversible cholestatic rat model in our recent study looking at the role of methylprednisolone in the ischemia reperfusion injury. Various techniques for creation of a reversible cholestatic model have been described. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.
This model can be used in various studies, such as looking at the changes in nutritional, physiological, pathological, histological and immunological changes in the gastrointestinal tract. This model can also be used to see the effects of cholestasis and various therapeutic interventions on major hepatic surgeries.

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.

가역 Cholestatic 쥐 모델의 생성

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and ...

연구

의학

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

무한 호라이즌 임팩터을 사용하여 일방적 경부 척수의 부상의 Contusive 모델

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction

MicroRNAs (miRNAs) are single stranded, non-coding RNA molecules that typically regulate gene expression at the post-transcriptional level by binding to partially complementary target sites in the 3&#39; untranslated region (UTR) of messenger RNA (mRNA), which reduces the mRNA&#39;s translation and stability. The miRNA expression profiles in various organs and tissues of mice have been investigated, but standard methods for the purification and quantification of miRNA in mouse kidney have not been available. We have established an effective and reliable method for extracting and evaluating miRNA expression in mouse kidney with renal interstitial fibrosis by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The protocol required five steps: (1) creation of sham and unilateral ureteral obstruction (UUO) mice; (2) extraction of kidney samples from the UUO mice; (3) extraction of total RNA, which includes miRNA, from the kidney samples; (4) complementary DNA (cDNA) synthesis with reverse transcription from miRNA; and (5) qRT-PCR using the cDNA. Using this protocol, we successfully confirmed that compared to the controls, the expression of miRNA-3070-3p was significantly increased and those of miRNA-7218-5p and miRNA-7219-5p were significantly decreased in the kidneys of a mouse model of renal interstitial fibrosis. This protocol can be used to determine the miRNA expression in the kidneys of mice with UUO.

We describe a method for evaluating the microRNA expression in the kidneys of mice with unilateral ureteral obstruction (UUO) by quantitative reverse-transcription polymerase chain reaction. This protocol is suitable for studying kidney microRNA expression profiles in mice with UUO and in the context of other pathological conditions.

일방적 인 요원 방해와 마우스 신장에서 microRNA 표현의 정량적 실시간 PCR 평가

MicroRNAs (miRNAs) are single stranded, non-coding RNA molecules that typically regulate gene expression at the post-transcriptional level by binding to ...

유전학

Nerve-sparing Mid-urethral Obstruction (NeMO) in Female Small Rodents

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal model to investigate bladder remodeling in pBOO are female rodents undergoing partial obstruction at the proximal urethra. Variability in the degree of obstruction and animal mortality are major concerns with proximal obstruction. Furthermore, dissecting around the proximal urethra and bladder neck jeopardizes bladder innervation.
We developed a nerve-sparing mid-urethral obstruction (NeMO) model for pBOO avoiding the disadvantages of the traditional model. We approached the urethra just inferior to the pubic symphysis, which obviated the need for laparotomy as well as for dissection in this area; also, the striated urethral sphincter remained untouched. We performed NeMO in female Sprague-Dawley rats (12 obstructions, 6 sham animals) as well as in female C57/bl6 mice (20 obstructions, 18 sham animals). After two weeks, we evaluated bladder function, bladder mass, and body mass.
We had no mortalities among obstructed- or sham-operated female rats; as described for the traditional proximal pBOO-method, we tied the suture around the proximal urethra and a temporarily placed 0.9 mm metal rod. NeMO induced an 85% increase in bladder mass after two weeks, average residual urine volume was 0.4 mL in partially obstructed rats while only 0.03 mL in sham animals. In mice, we tested 3 sizes of cannulas that we placed along the urethra when tying the suture. We found that using a 27-gauge cannula resulted in over 50% animal mortality; placing the 25-gauge cannula did not yield the desired response in increasing bladder mass; utilizing a 26-gauge cannula yielded favorable results with minimal animal mortality (1/8) yet a significant 2-fold increase in bladder mass.

Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents

Traditional modeling of partial bladder outlet obstruction in rodents is fraught with animal mortality. A denervation injury from dissection around the proximal urethra and bladder neck is also of major concern. We developed and evaluated a safe and reliable mid-urethral obstruction model, avoiding the shortcomings of the traditional model.

여성 작은 설치류에 신경을 살려주는 중간 요도 폐색 (네모)

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal ...

Detrusor Underactivity Model in Rats by Conus Medullaris Transection

The goal of the presented protocol was to establish a detrusor underactivity (DU) model in the rat through conus medullaris transection. Laminectomy was performed in a total of 40 female Wistar rats (control group: 10 rats; test group: 30 rats) weighing 200&#8211;220 g, and the conus medullaris was transected at the L4&#8210;L5 level in the test group. All the rats were housed and fed under the same environmental conditions for six weeks. In the test group, urine voiding was performed twice daily for six weeks, and mean residual urine volume was recorded. A cystometrogram was performed in both groups. Maximum cystometric capacity (MCC), detrusor opening pressure (DOP), and compliance of the bladder were recorded and calculated. The test group showed significant urinary retention after the surgery, both during and after the spinal shock. However, no abnormality was observed in the control group. When compared to the control group, the MCC and compliance of bladder in the test group was significantly higher than that of the test group (3.24 &#177; 2.261 mL versus 1.04 &#177; 0.571 mL; 0.43 &#177; 0.578 mL/cmH2O versus 0.032 &#177; 0.016 mL/cmH2O), whereas DOP in the test group was lower than control (20.28 &#177; 14.022 cmH2O versus 35 &#177; 13.258 cmH2O). This method of establishing an animal model of DU by the conus medullaris transection offers an excellent opportunity to understand DU&#8217;s pathophysiology in a better manner.

We present a method for establishing a detrusor underactivity model by conus medullaris transection in rats. Detrusor underactivity was successfully stimulated in these animals. The model can be used for studying urinary tract function.

원추 수질 절개에 의한 쥐의 배뇨근 과소 모델

The goal of the presented protocol was to establish a detrusor underactivity (DU) model in the rat through conus medullaris transection. Laminectomy was ...

신경과학

An Effective Mouse Model of Unilateral Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models are the only available resources that mimic the complexities of the IRI-associated damage encountered in vivo. This paper describes an effective mouse model of unilateral renal IRI that delivers highly reproducible data. Ischemia is induced by occluding the right renal pedicle for 30 min followed by reperfusion. In addition to the surgical procedure, a sequential overview of the expected physiological and histopathological changes following renal IRI will be provided by comparing data from seven different reperfusion times (4 h, 8 h, 16 h, 1 day, 2 days, 4 days, and 7 days). Critical data for planning experiments ahead, such as mean surgical time, average anesthetic consumption, and body weight changes over time, will be shared. This work will help researchers implement a reliable renal IRI model and select the appropriate reperfusion time that aligns with their intended investigative goals.

Renal ischemia-reperfusion injury is associated with high morbidity and mortality in hospitalized patients. Here, we present a simple and effective mouse model of unilateral renal ischemia-reperfusion injury and provide a sequential overview of representative pathological changes observed in the kidney.

일방적 인 신장 허혈 - 재관류 손상의 효과적인 마우스 모델

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models ...

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.

2광자 현미경을 사용하여 만성 일방적 요관 폐쇄가 사구체 과정에 미치는 영향을 정량화

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

An Immature Murine Model of Reversible Unilateral Ureteral Obstruction

Unilateral ureteral obstruction (UUO) is a common cause of chronic kidney disease (CKD), leading to the progression of renal interstitial fibrosis and ultimately resulting in irreversible kidney damage. The alleviation of UUO is crucial. Several animal models of reversible unilateral ureteral obstruction (RUUO) have been established in the literature, enabling the observation of structural changes and functional damage while also simulating physiological and pathophysiological changes following the relief of ureteral obstruction. In this study, a reversible obstruction model was established in the unilateral murine ureter using a silicone tube. Significant renal damage was observed prior to obstruction relief, with partial recovery noted afterward. Unlike UUO, this model prevents progressive hydronephrosis, leading to distinct pathological outcomes. This simple surgical procedure demonstrates a high success rate and holds promise as a classical model for investigating reversible obstructive nephropathy and potential treatments for renal interstitial fibrosis. Furthermore, it provides a practical platform for studying the mechanisms of recovery from obstructive nephropathy, renal cell regeneration, and tissue remodeling.

The present protocol describes a step-by-step, reproducible model of unilateral ureteral obstruction.

가역적 일방적 요관 폐색의 미성숙한 쥐 모델(An Immature Murine Model of Reversible Unilateral Ureteral Obstruction)

Unilateral ureteral obstruction (UUO) is a common cause of chronic kidney disease (CKD), leading to the progression of renal interstitial fibrosis and ...

Unilateral Ureteral Obstruction Model for Investigating Kidney Interstitial Fibrosis

Kidney fibrosis is the final pathological outcome of progressive chronic kidney disease (CKD). The unilateral ureteral obstruction (UUO) model is widely used to elucidate the molecular and cellular mechanisms underlying kidney interstitial fibrosis and to identify potential therapeutic targets. This model is established in mice through surgical ligation of a unilateral ureter, a procedure that is relatively straightforward and easy to perform. However, the UUO mouse model is known to exhibit significant variability and inconsistency, influenced by factors such as mouse strain, age, sex, anesthesia type, duration of surgery, body temperature during the procedure, the operator's surgical skills, feeding conditions, and the overall health status of the mice. Variations in surgical techniques, suture placement, and the duration of obstruction contribute to the variability in outcomes. Additionally, inconsistent sampling of obstructed kidneys further increases variability in the assessment of kidney fibrosis. This study outlines the process of developing the UUO mouse model and evaluating interstitial fibrosis, discusses the technical challenges contributing to the model's unpredictability, and proposes potential solutions. These insights aim to establish a more standardized and universally applicable approach for investigating kidney fibrosis.

This protocol describes a method for inducing unilateral ureteral obstruction (UUO) in mice to study the progression of tissue fibrosis in obstructive nephropathy. It includes surgical procedures, post-operative care, and methods for fibrosis assessment.

신장 간질성 섬유증(Kidney Interstitial Fibrosis) 조사를 위한 일측성 요관 폐쇄 모델(Unilateral Ureteral Obstruction Model for Investigating Kidney Interstitial Fibrosis)

Kidney fibrosis is the final pathological outcome of progressive chronic kidney disease (CKD). The unilateral ureteral obstruction (UUO) model is widely ...

Unilateral Ureteral Obstruction Murine Model: A Surgical Procedure to Obstruct the Urinary Flow from a Kidney in Mouse Models

Source: Yanai, K. et al. <a href="https://www.jove.com/video/61383" target="_blank">Quantitative real-time PCR evaluation of microRNA expressions in mouse kidney with unilateral ureteral obstruction.</a> J. Vis. Exp. (2020)
This video demonstrates the procedure to generate a unilateral ureteral obstruction mouse model. The procedure involves cutting one of the ureters of the mouse to obstruct urine flow.

편측성 요관 폐색 쥐 모델: 마우스 모델에서 신장의 소변 흐름을 막는 수술 절차

Source: Yanai, K. et al. Quantitative real-time PCR evaluation of microRNA expressions in mouse kidney with unilateral ureteral obstruction. J. Vis. Exp. ...