Name: Unilateral Ureteral Obstruction Model for Investigating Kidney Interstitial Fibrosis (Video) | JoVE Journal
Uploaded: 2025-04-25T14:00:00.000+00:00
Duration: 4 min 37 s
Description: Learn the unilateral ureteral obstruction model for investigating kidney interstitial fibrosis in mice. This protocol enables researchers to study macrophage interactions and identify therapeutic targets.

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5 minutes

This work was supported by National Science Foundation of China Grants (82370686/2024YFA1107704), Jiangsu Specially-Appointed Professor Grant, Nanjing Science and Technology Innovation Project, Jiangsu Province Hospital High-level Talent Cultivation Program (Phase I) (CZ0121002010037), Natural Science Foundation of Jiangsu Province (BK20240055), and Jiangsu Medical Innovation Team to JR; Jiangsu Province Hospital (the First Affiliated Hospital with Nanjing Medical University) Clinical Capacity Enhancement Project (JSPH-MA-2023-4), Priority Academic Program Development of Jiangsu Higher Education Institutions (China) and National Natural Science Foundation of China (81970639/82151320) to HM.

All animal procedures are conducted in accordance with agency guidelines and approved by the Institutional Animal Ethics Committee of Nanjing Medical University. To eliminate sex and strain differences and ensure comparability of results, only male CD1 mice aged 8-10 weeks and weighing 22-25 g are used. The details of the reagents and equipment used in this study are listed in the Table of Materials.
1. Animal and instrument preparation
<ol>
	<li>Autoclave all surgical instruments before surgery.</li>
	<li>Weigh and anesthetize the mice (following institutionally approved protocols). Properly connect the inhalation anesthesia device and place the mice in the induction chamber with 2% isoflurane mixed with oxygen at 1-2 L/min. After confirming anesthetic induction (e.g., loss of righting reflex), place the mouse&#39;s nose into the nose cone with a constant supply of anesthesia. Administer buprenorphine (50 &#181;g/kg) by subcutaneous injection as pre-operative analgesia. 
	NOTE: Regularly check the respiration rate and assess the level of anesthesia using the pedal reflex of the mice. Adjust the isoflurane as appropriate.</li>
	<li>Place the mouse on an external&#160;heat source&#160;in a supine position with the head and neck extended to keep the airway open. Secure the paws with low-adhesive tape.</li>
	<li>Use a thermometer to monitor the ambient temperature and maintain it at 37-38 &#176;C. Insert a rectal probe and keep the core body temperature of the mice around 36.5-37 &#176;C during the procedure.</li>
	<li>Apply ophthalmic ointment to the eyes to prevent corneal xerosis injury.</li>
	<li>Remove hair over the surgical area with hair removal cream or clippers and disinfect it three times using betadine solution and 75% ethanol.</li>
</ol>
2. Surgical procedure
NOTE: Once the body temperature stabilizes at the set point and the toe pinch reflex is absent, initiate the following surgical procedures.
<ol>
	<li>Use a surgical blade to make a vertical surgical incision of approximately 1- 1.5 centimeters from the bladder to the lower-left edge of the ribs to expose the abdominal cavity.</li>
	<li>Move the intestine to the right side of the abdominal cavity using a cotton swab moistened with sterile normal saline to expose the left kidney, located below the back of the spleen and adjacent to the spine.</li>
	<li>Use curved iris forceps to clear the fat and connective tissue surrounding the left ureter. After gentle isolation, ligate the left ureter close to the kidney&#39;s lower pole using a 4/0 surgical suture. Perform a second ligation of the left ureter between the first ligation and the bladder. 
	NOTE: To minimize variability among the mice, ensure that the ligation position remains consistent across all subjects. Use the left-hand forceps to place the suture into the tips of the right-hand forceps and slide the suture back under the ureter17. Avoid including excess fat and connective tissue in the ligation, as this may lead to incomplete kidney obstruction. Achieve this by gently stroking the ureter with dry cotton swabs.</li>
	<li>Carefully reposition the intestines into the peritoneal cavity. Suture the muscle and skin layers separately using&#160;4/0 surgical sutures. 
	NOTE: Suturing the skin and muscle layers together may cause suture wound rupture and abdominal viscera damage.</li>
	<li>When operating on multiple mice, use a fresh autoclaved surgical instrument&#160;before proceeding to the next mouse.</li>
</ol>
3. Post-surgical care and monitoring
<ol>
	<li>Inject 0.5 mL of warm normal saline intraperitoneally to prevent dehydration.</li>
	<li>Place the mice back into a clean cage until they regain full consciousness.</li>
	<li>Administer buprenorphine (50 &#181;g/kg) by subcutaneous injection every 8 h for 3 days post-surgery. Monitor the mice daily for signs of lack of grooming, decreased eating, and abnormal posture.</li>
</ol>
4. Post-operative assessments
<ol>
	<li>Histology
	<ol>
		<li>Euthanize the mice with an overdose of isoflurane (following institutionally approved protocols). Harvest the obstructed and contralateral non-obstructed kidneys18 after euthanasia.</li>
		<li>Use the largest cross-sectional area of the kidney section for histological examination.</li>
		<li>Fix kidney tissues in 4% paraformaldehyde and cut the obtained samples into 4 &#181;m sections.</li>
		<li>Stain kidney sections with periodic acid-Schiff (PAS), Masson&#39;s trichrome staining (MTS), or Sirius red staining to detect renal tubular injury and kidney fibrosis.</li>
		<li>Select ten random fields of the kidney sections under a light microscope at a magnification of 200&#215;.</li>
		<li>Calculate the proportion of the collagen-positive blue area using ImageJ software.</li>
	</ol>
	</li>
	<li>Western blot
	<ol>
		<li>Extract approximately 20 mg of kidney tissue from the same pole of UUO kidneys using radioimmunoprecipitation assay (RIPA) buffer18.</li>
		<li>Quantify the protein concentration using a bicinchoninic acid assay19.</li>
		<li>Load equal amounts of protein samples onto 4%-10% Bis-Tris gels for electrophoresis and transfer them to polyvinylidene difluoride (PVDF) membranes.</li>
		<li>Block the PVDF membranes with 5% milk in Tris-Buffered Saline Tween (TBST) and incubate them with primary antibodies overnight at 4 &#176;C.</li>
		<li>Wash the PVDF membranes with TBST and incubate them with secondary antibodies at room temperature for 1 h.</li>
		<li>Detect protein levels using an enhanced chemiluminescence (ECL) method and analyze them using ImageJ software, with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the loading control.</li>
	</ol>
	</li>
	<li>Renal Function
	<ol>
		<li>Anesthetize the mice with inhaled isoflurane (see step 1.2).</li>
		<li>Collect 0.2-0.3 mL of blood retro-orbitally after anesthesia.</li>
		<li>Centrifuge blood samples at 3,000 x g for 15 min at room temperature.</li>
		<li>Measure serum creatinine and blood urea nitrogen (BUN) using an automatic dry chemical analyzer to monitor renal function.</li>
	</ol>
	</li>
</ol>

<ol><li>Bello, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+update+on+the+global+disparities+in+kidney+disease+burden+and+care+across+world+countries+and+regions%5BTitle%5D)+AND+%22Lancet+Glob+Health%22%5BJournal%5D)">An update on the global disparities in kidney disease burden and care across world countries and regions.</a> Lancet Glob Health. 12 (3), e382-e395 (2024).</li>
<li>Klahr, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Obstructive+nephropathy%5BTitle%5D)+AND+%22Intern+Med%22%5BJournal%5D)">Obstructive nephropathy.</a> Intern Med. 39 (5), 355-361 (2000).</li>
<li>Lovisa, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Epithelial-to-mesenchymal+transition+induces+cell+cycle+arrest+and+parenchymal+damage+in+renal+fibrosis%5BTitle%5D)+AND+%22Nat+Med%22%5BJournal%5D)">Epithelial-to-mesenchymal transition induces cell cycle arrest and parenchymal damage in renal fibrosis.</a> Nat Med. 21 (9), 998-1009 (2015).</li>
<li>Grande, M. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Snail1-induced+partial+epithelial-to-mesenchymal+transition+drives+renal+fibrosis+in+mice+and+can+be+targeted+to+reverse+established+disease%5BTitle%5D)+AND+%22Nat+Med%22%5BJournal%5D)">Snail1-induced partial epithelial-to-mesenchymal transition drives renal fibrosis in mice and can be targeted to reverse established disease.</a> Nat Med. 21 (9), 989-997 (2015).</li>
<li>Kang, H. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Defective+fatty+acid+oxidation+in+renal+tubular+epithelial+cells+has+a+key+role+in+kidney+fibrosis+development%5BTitle%5D)+AND+%22Nat+Med%22%5BJournal%5D)">Defective fatty acid oxidation in renal tubular epithelial cells has a key role in kidney fibrosis development.</a> Nat Med. 21 (1), 37-46 (2015).</li>
<li>Lebleu, V. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Origin+and+function+of+myofibroblasts+in+kidney+fibrosis%5BTitle%5D)+AND+%22Nat+Med%22%5BJournal%5D)">Origin and function of myofibroblasts in kidney fibrosis.</a> Nat Med. 19 (8), 1047-1053 (2013).</li>
<li>Yang, L., Besschetnova, T. Y., Brooks, C. R., Shah, J. V., Bonventre, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Epithelial+cell+cycle+arrest+in+g2%2Fm+mediates+kidney+fibrosis+after+injury%5BTitle%5D)+AND+%22Nat+Med%22%5BJournal%5D)">Epithelial cell cycle arrest in g2/m mediates kidney fibrosis after injury.</a> Nat Med. 16 (5), 535-543 (2010).</li>
<li>Kuppe, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Decoding+myofibroblast+origins+in+human+kidney+fibrosis%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Decoding myofibroblast origins in human kidney fibrosis.</a> Nature. 589 (7841), 281-286 (2021).</li>
<li>Ren, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+transcription+factor+twist1+in+the+distal+nephron+but+not+in+macrophages+propagates+aristolochic+acid+nephropathy%5BTitle%5D)+AND+%22Kidney+Int%22%5BJournal%5D)">The transcription factor twist1 in the distal nephron but not in macrophages propagates aristolochic acid nephropathy.</a> Kidney Int. 97 (1), 119-129 (2020).</li>
<li>Perales-Quintana, M. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Metabolomic+and+biochemical+characterization+of+a+new+model+of+the+transition+of+acute+kidney+injury+to+chronic+kidney+disease+induced+by+folic+acid%5BTitle%5D)+AND+%22PeerJ%22%5BJournal%5D)">Metabolomic and biochemical characterization of a new model of the transition of acute kidney injury to chronic kidney disease induced by folic acid.</a> PeerJ. 7, e7113(2019).</li>
<li>Kim, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Skeletal+myopathy+in+CKD%3A+A+comparison+of+adenine-induced+nephropathy+and+5%2F6+nephrectomy+models+in+mice%5BTitle%5D)+AND+%22Am+J+Physiol+Renal+Physiol%22%5BJournal%5D)">Skeletal myopathy in CKD: A comparison of adenine-induced nephropathy and 5/6 nephrectomy models in mice.</a> Am J Physiol Renal Physiol. 321 (1), F106-F119 (2021).</li>
<li>Tan, R. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+optimized+5%2F6+nephrectomy+mouse+model+based+on+unilateral+kidney+ligation+and+its+application+in+renal+fibrosis+research%5BTitle%5D)+AND+%22Ren+Fail%22%5BJournal%5D)">An optimized 5/6 nephrectomy mouse model based on unilateral kidney ligation and its application in renal fibrosis research.</a> Ren Fail. 41 (1), 555-566 (2019).</li>
<li>Fu, Y., Xiang, Y., Wei, Q., Ilatovskaya, D., Dong, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rodent+models+of+aki+and+AKI-CKD+transition%3A+An+update+in+2024%5BTitle%5D)+AND+%22Am+J+Physiol+Renal+Physiol%22%5BJournal%5D)">Rodent models of aki and AKI-CKD transition: An update in 2024.</a> Am J Physiol Renal Physiol. 326 (4), F563-F583 (2024).</li>
<li>Kramann, R., Menzel, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mouse+models+of+kidney+fibrosis%5BTitle%5D)+AND+%22Methods+Mol+Biol%22%5BJournal%5D)">Mouse models of kidney fibrosis.</a> Methods Mol Biol. 2299, 323-338 (2021).</li>
<li>Chevalier, R. L., Forbes, M. S., Thornhill, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ureteral+obstruction+as+a+model+of+renal+interstitial+fibrosis+and+obstructive+nephropathy%5BTitle%5D)+AND+%22Kidney+Int%22%5BJournal%5D)">Ureteral obstruction as a model of renal interstitial fibrosis and obstructive nephropathy.</a> Kidney Int. 75 (11), 1145-1152 (2009).</li>
<li>Nørregaard, R., Mutsaers, H. aM., Frøkiær, J., Kwon, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Obstructive+nephropathy+and+molecular+pathophysiology+of+renal+interstitial+fibrosis%5BTitle%5D)+AND+%22Physiol+Rev%22%5BJournal%5D)">Obstructive nephropathy and molecular pathophysiology of renal interstitial fibrosis.</a> Physiol Rev. 103 (4), 2827-2872 (2023).</li>
<li>Thornhill, B. A., Chevalier, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Variable+partial+unilateral+ureteral+obstruction+and+its+release+in+the+neonatal+and+adult+mouse%5BTitle%5D)+AND+%22Methods+Mol+Biol%22%5BJournal%5D)">Variable partial unilateral ureteral obstruction and its release in the neonatal and adult mouse.</a> Methods Mol Biol. 886, 381-392 (2012).</li>
<li>Ren, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Twist1+in+infiltrating+macrophages+attenuates+kidney+fibrosis+via+matrix+metallopeptidase+13-mediated+matrix+degradation%5BTitle%5D)+AND+%22J+Am+Soc+Nephrol%22%5BJournal%5D)">Twist1 in infiltrating macrophages attenuates kidney fibrosis via matrix metallopeptidase 13-mediated matrix degradation.</a> J Am Soc Nephrol. 30 (9), 1674-1685 (2019).</li>
<li>Smith, P. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Measurement+of+protein+using+bicinchoninic+acid%5BTitle%5D)+AND+%22Anal+Biochem%22%5BJournal%5D)">Measurement of protein using bicinchoninic acid.</a> Anal Biochem. 150 (1), 76-85 (1985).</li>
<li>Puri, T. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Chronic+kidney+disease+induced+in+mice+by+reversible+unilateral+ureteral+obstruction+is+dependent+on+genetic+background%5BTitle%5D)+AND+%22Am+J+Physiol+Renal+Physiol%22%5BJournal%5D)">Chronic kidney disease induced in mice by reversible unilateral ureteral obstruction is dependent on genetic background.</a> Am J Physiol Renal Physiol. 298 (4), F1024-F1032 (2010).</li>
<li>Falke, L. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Age-dependent+shifts+in+renal+response+to+injury+relate+to+altered+BMP6%2FCTGF+expression+and+signaling%5BTitle%5D)+AND+%22Am+J+Physiol+Renal+Physiol%22%5BJournal%5D)">Age-dependent shifts in renal response to injury relate to altered BMP6/CTGF expression and signaling.</a> Am J Physiol Renal Physiol. 311 (5), F926-F934 (2016).</li>
<li>Song, L., Shi, S., Jiang, W., Liu, X., He, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Protective+role+of+propofol+on+the+kidney+during+early+unilateral+ureteral+obstruction+through+inhibition+of+epithelial-mesenchymal+transition%5BTitle%5D)+AND+%22Am+J+Transl+Res%22%5BJournal%5D)">Protective role of propofol on the kidney during early unilateral ureteral obstruction through inhibition of epithelial-mesenchymal transition.</a> Am J Transl Res. 8 (2), 460-472 (2016).</li>
<li>Saad, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reno-protective+effect+of+protocatechuic+acid+is+independent+of+sex-related+differences+in+murine+model+of+UUO-induced+kidney+injury%5BTitle%5D)+AND+%22Pharmacol+Rep%22%5BJournal%5D)">Reno-protective effect of protocatechuic acid is independent of sex-related differences in murine model of UUO-induced kidney injury.</a> Pharmacol Rep. 76 (1), 98-111 (2024).</li>
<li>Goorani, S., Khan, A. H., Mishra, A., El-Meanawy, A., Imig, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Kidney+injury+by+unilateral+ureteral+obstruction+in+mice+lacks+sex+differences%5BTitle%5D)+AND+%22Kidney+Blood+Press+Res%22%5BJournal%5D)">Kidney injury by unilateral ureteral obstruction in mice lacks sex differences.</a> Kidney Blood Press Res. 49 (1), 69-80 (2024).</li>
<li>La Russa, D., Barberio, L., Marrone, A., Perri, A., Pellegrino, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Caloric+restriction+mitigates+kidney+fibrosis+in+an+aged+and+obese+rat+model%5BTitle%5D)+AND+%22Antioxidants+%28Basel%29%22%5BJournal%5D)">Caloric restriction mitigates kidney fibrosis in an aged and obese rat model.</a> Antioxidants (Basel). 12 (9), 1778(2023).</li>
<li>Wei, Q., Dong, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mouse+model+of+ischemic+acute+kidney+injury%3A+Technical+notes+and+tricks%5BTitle%5D)+AND+%22Am+J+Physiol+Renal+Physiol%22%5BJournal%5D)">Mouse model of ischemic acute kidney injury: Technical notes and tricks.</a> Am J Physiol Renal Physiol. 303 (11), F1487-F1494 (2012).</li>
<li>Hammad, F. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+long-term+renal+effects+of+short+periods+of+unilateral+ureteral+obstruction%5BTitle%5D)+AND+%22Int+J+Physiol+Pathophysiol+Pharmacol%22%5BJournal%5D)">The long-term renal effects of short periods of unilateral ureteral obstruction.</a> Int J Physiol Pathophysiol Pharmacol. 14 (2), 60-72 (2022).</li>
<li>Chevalier, R. L., Thornhill, B. A., Wolstenholme, J. T., Kim, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Unilateral+ureteral+obstruction+in+early+development+alters+renal+growth%3A+Dependence+on+the+duration+of+obstruction%5BTitle%5D)+AND+%22J+Urol%22%5BJournal%5D)">Unilateral ureteral obstruction in early development alters renal growth: Dependence on the duration of obstruction.</a> J Urol. 161 (1), 309-313 (1999).</li>
</ol>

The authors declare no conflict of interest.

A comprehensive procedure for establishing the UUO model, a widely used approach for investigating kidney interstitial fibrosis, is provided. Additionally, the identification and assessment of the model, including evaluations of renal function and histological alterations, are demonstrated. The variables contributing to the model's heterogeneity and modifiable technical factors are discussed.
Susceptibility to UUO varies significantly based on age, sex, and mouse strain. Compared to C57BL/6 mi...

Chronic kidney disease (CKD) affects over 10% of the global population, and its prevalence is increasing1. Various urinary tract conditions, including congenital anatomical anomalies, nephrolithiasis, prostatic hyperplasia, and bladder tumors, can lead to ureteral obstruction&#178;. As a result, the unilateral ureteral obstruction (UUO) mouse model is a key tool for identifying new mechanisms of kidney interstitial fibrosis, understanding disease progression, and evaluating potential treatment strategies. It has been widely used to investigate the origin of myofibroblasts, (myo)fibroblast subclusters, tubular cell metabolism, and cell cycle arrest, partial epithelial-mesenchymal transition, and other related processes3,4,5,6,7,8.
In addition to UUO-induced kidney interstitial fibrosis, other commonly used rodent models of kidney interstitial fibrosis include toxin-induced models, such as those using aristolochic acid, folic acid, and adenine, as well as surgically induced models like 5/6 nephrectomy and ischemia-reperfusion injury (IRI). The UUO model offers several advantages over alternative kidney fibrosis models. For example, toxin-induced kidney fibrosis requires a relatively long modeling period (approximately 1-2 months), and its toxic side effects on other organs can complicate the investigation of fibrosis mechanisms9,10,11. Surgically induced models, such as 5/6 nephrectomy, can lead to significant kidney bleeding and infection, increasing the risk of post-operative mortality. Additionally, the extent of induced interstitial fibrosis is directly correlated with the volume of resected kidney tissue, making it challenging to consistently reproduce the same degree of fibrosis in each mouse12.
The renal IRI model is a primary method for inducing acute kidney injury to CKD and has significant clinical relevance. The severity of fibrosis can be modulated by adjusting ischemic time and body temperature; however, compared to the UUO model, it is more surgically complex, and the induction of interstitial fibrosis requires a longer duration13. Compared to these models, the UUO model has several advantages, including a short modeling duration, minimal variability, repeatability, and a relatively simple surgical procedure. The UUO mouse model, which does not involve toxins, is created by ligating one ureter, leading to obstructive nephropathy within two weeks. This results in hydronephrosis, tubular dilatation, and interstitial fibrosis, closely resembling the pathological process observed in humans14. The severity of fibrosis -- mild, moderate, or severe -- can be controlled by adjusting the experiment&#39;s duration.
Although the UUO mouse model is simpler to perform than other insult-induced models for investigating CKD, several factors can significantly affect its stability. These factors include mouse strain, age, sex, type of anesthesia, surgery duration, body temperature during surgery, the surgical skills of the operator, and the feeding conditions and health status of the mice15,16.
Minimizing surgical stress and infection while performing the procedure in a steady and organized manner under anesthesia is essential for creating a reproducible UUO mouse model. Additionally, research on the mechanisms and potential therapeutic targets of CKD can be compromised by inexperienced operators, leading to increased mouse loss and greater model heterogeneity. To address these challenges, key technical aspects of the surgical process -- before, during, and after the procedure -- are outlined, highlighting critical issues that require attention. Furthermore, the evaluation methodology for the UUO mouse model is detailed to provide researchers with a consistent and reliable approach.

<table><tbody><tr><td>1 mL Syringe</td><td>Mingankang</td><td>/</td><td></td></tr><tr><td>4/0 surgical suture</td><td>Jinhuan Medical</td><td>F401</td><td></td></tr><tr><td>75% Ethanol</td><td>Lircon</td><td>6303060031</td><td></td></tr><tr><td>Anesthesia Air Pump</td><td>RWD Life Science</td><td>R510-29</td><td></td></tr><tr><td>Anesthesia Induction Chambers</td><td>RWD Life Science</td><td>V102-V</td><td></td></tr><tr><td>Animal hair clipper</td><td>Jinke</td><td>/</td><td></td></tr><tr><td>Betadine solution</td><td>Lircon</td><td>6303030036</td><td></td></tr><tr><td>Buprenorphine (analgesic)</td><td>RWD Life Science</td><td>/</td><td></td></tr><tr><td>Curved iris forceps</td><td>jinke</td><td>/</td><td></td></tr><tr><td>Electronic heat pad</td><td>Reptizoo</td><td>AHM23</td><td></td></tr><tr><td>fine straight forceps</td><td>Jinke</td><td>/</td><td></td></tr><tr><td>Gas Filter Canister</td><td>RWD Life Science</td><td>R510-31-6</td><td></td></tr><tr><td>Gauze Pads</td><td>Winner Medical</td><td>601-026576</td><td></td></tr><tr><td>Iris Scissors</td><td>Jinke</td><td>/</td><td></td></tr><tr><td>Isoflurane (anesthetic)&amp;nbsp;</td><td>RWD Life Science</td><td>R510-22-10</td><td></td></tr><tr><td>Multi-output Animal Anesthesia Machine</td><td>RWD Life Science</td><td>R550IE</td><td></td></tr><tr><td>Needle holder</td><td>jinke</td><td>/</td><td></td></tr><tr><td>Ophthalmic ointment</td><td>Dechra NDC&amp;nbsp;</td><td>17033-211-38</td><td></td></tr><tr><td>Sterile Cotton swab</td><td>Winner Medical</td><td>601-015213</td><td></td></tr><tr><td>Sterile saline</td><td>Shimen</td><td>H20066533</td><td></td></tr></tbody></table>

unilateral ureteral obstruction model for investigating kidney interstitial fibrosis

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.

Histology 
Periodic acid-Schiff (PAS) staining revealed tubular dilation, loss of brush borders, cast formation, and tubular epithelial swelling. Masson's trichrome and Sirius red staining showed interstitial fibrosis following UUO, in contrast to the normal compact tubules with discernible lumens observed in the sham group. The degree of renal interstitial fibrosis, indicated by blue areas in Masson's trichrome staining and red areas in Sirius red staining, increased in a time-dependent manner (<stro...

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.

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

unilateral-ureteral-obstruction-model-for-investigating-kidney

Research

JoVE Journal

Medicine

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.

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

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

Mouse Kidney Transplantation: Models of Allograft Rejection

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.

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

This study presents a highly reproducible large animal model of renal ischemia-reperfusion injury in swine using temporary percutaneous bilateral balloon-catheter occlusion of the renal arteries for 60 min and reperfusion for 24 h.

A Large Animal Model for Acute Kidney Injury by Temporary Bilateral Renal Artery Occlusion

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.

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

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

An Immature Murine Model of Reversible Unilateral Ureteral Obstruction

This paper provides detailed instructions to enable investigators to perform reversible unilateral ureteric obstruction in mice using a standardized model that allows analysis of the long-term structural and functional outcomes after the reversal of prolonged ureteric obstruction.

A Mouse Model to Evaluate the Long-Term Structural and Functional Outcomes after the Reversal of Prolonged Unilateral Ureteric Obstruction

This article presents an ischemic CKD model using unilateral renal IRI with contralateral nephrectomy performed one day prior to sample collection; this protocol enables serum-based functional assessment, high survival, and reproducible CKD pathology at 28 and 42 days, providing a versatile platform for mechanistic and interventional studies.

Mouse Model of Acute to Chronic Kidney Disease Transition Induced by Renal Ischemia/Reperfusion Injury

We describe a method to produce an animal model of liver fibrosis in the rat, and assess the degree of fibrosis by histological examination of the liver. The model can be used to study the development of liver disease as well as to test the efficacy of potential anti-fibrotic agents.

The Dimethylnitrosamine Induced Liver Fibrosis Model in the Rat

Learn the unilateral ureteral obstruction model for investigating kidney interstitial fibrosis in mice. This protocol enables researchers to study macrophage interactions and identify therapeutic targets.

Unilateral Ureteral Obstruction Model for Investigating Kidney Interstitial Fibrosis

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