The authors would like to acknowledge the support of NIH/NIDDK K12 DK100014 (Lan Lu), the Case Comprehensive Cancer Center (NIH/NCI P30 CA43703), and the Clinical and Translation Science Collaborative of Cleveland (NIH/NCATS UL1 TR000439).

所有动物研究都是根据Case Western Reserve大学批准的机构动物护理和使用委员会（IACUC）方案进行的。下述外科手术的持续时间约为45-60分钟。每个时间点MRI程序本身约为15分钟。 麻醉<ol><li>在异氟烷麻醉室内麻醉大鼠，其设定为与氧气混合的2％异氟烷，以便在腹膜内施用可注射麻醉之前促进动物处理和约束。 </li><li>暴露于异氟烷3-5分钟后，检查动物是否被麻醉，并且对脚趾没有反应。 </li><li>进一步镇静大鼠腹膜内注射赛拉嗪和氯胺酮:75mg / kg氯胺酮/ 10mg / kg甲苯噻嗪。当进行腹膜内注射时，拉回针头以确保肠或其他重要器官的部分未被穿刺。 </li><li>一世在切口部位皮下注射2 mg / kg布比卡因以提供局部疼痛缓解。 </li></ol> 2.手术区域的准备<ol><li>在外科手术器械和用品用于手术之前灭菌并放置在外科手术垫上进行无菌处理。大多数仪器和耗材可以进行高压灭菌和重新使用。 </li><li>使用无菌手套进行所有外科手术。 </li><li>使用电动剃须刀将皮毛从动物的右侧剃掉。将动物从肋骨的底部剃除到后腿的顶部，为切口提供一个大的无毛的区域。 </li><li>将动物放在无菌手术垫上，以将消毒区域与周围区域隔离。 </li><li>用消毒剂如聚维酮碘或betadine擦洗皮肤。开始在手术中心的擦洗，并以圆形的方式移动到外面。用新的聚维酮碘或betadine擦拭至少重复三次。 </LI&gt; <li>用70％酒精拭子擦洗手术部位，直到皮肤清晰，因为如果吸收碘可能会有毒性。 </li></ol>手术程序<ol><li>在无菌条件下维持此程序。 </li><li>将麻醉动物置于左侧卧位的暖暖床上，右侧腹部朝上。 注意:必须注意将动物的核心体温保持在35-37°C以防止体温过低。这个升温床也应根据需要进行灭菌以维持无菌条件。 </li><li>感觉为肋骨笼，并使用无菌尺寸10手术刀刀片从肋骨底部开始，做一个小2-3厘米的右背腹膜后切口。 </li><li>沿切口两侧纵向放置无菌纱布。 </li><li>解剖皮下组织，脂肪和肌肉，以便可视化和进入腹腔。使用弯刀Mayo剪刀可以进一步穿透配给伤口并切割厚厚的组织。 </li><li>一旦肝脏清晰可见，便可使用钝镊子将肝脏向上收缩。 </li><li>另一方面使用另一对钝钳，暴露右肾，使其位于腹腔外。 </li><li>使用左手的手指和拇指将肾脏固定就位。用右手慢慢稳定地将0.1ml UTI89 大肠杆菌溶液（浓度在1×10 8 -1×10 9 ）从无菌注射器注入肾盂（其呈现为白色气泡） 15 。 注意:按照参考文献15所述准备细菌滴度。 </li><li>在针上放置一条可吸收的止血钳，以防止接种物流入腹膜。慢慢将针从肾盂拉出。 注意:注意不要刺穿或感染周围的组织导致非目标感染症状和/或并发症。 </li><li>使用较大的注射器，用生理盐水彻底冲洗肾脏，然后放回腹腔。 </li></ol>缝合线注意:将被埋在组织中的缝线应为4-0不可吸收的编织缝线。可吸收或单丝缝线可用于身体表面。 <ol><li>将缝合线均匀并尽可能靠近组织边缘，以防止血流阻塞;通常距离边缘不超过0.3厘米。 </li><li>抓住皮肤并使用一对细齿镊略微转动，并将针头保持器旋转到旋前的位置，以准备刺穿皮肤。 </li><li>通过放置手腕旋转针头并将其穿过皮肤，将针头穿过皮肤的全部厚度。 </li><li>对最接近执行过程的个人的皮肤边缘重复此过程。 </li><li>紧缩缝线足以抵抗组织边缘。任何更紧的会阻塞血液供应，伤口愈合缓慢，并可能导致开裂。 </li><li>使用方形结缝合缝合线，就好像它是一个简单的中断线迹，只不过只有短绞线被切割，留下约3-4毫米的尾巴。 </li><li>在第一针被束缚之后，准备放置距离第一针约3毫米的第二针，然后继续缝合线。 </li><li>一旦达到切口的末端，不要将最后一个针头完全拉过来。相反，使用在此处将针保持器保持的环作为短股线，以便将缝合线封闭件的远端扣紧。 </li><li>使用仪器领带，使用方形结缝合缝线。这导致3股从远端的完成的结粘连。 </li></ol> 5.动物恢复<ol><li>手术后腹膜内注射2.1 mg / kg育亨宾逆转麻醉。一个3-5小时后，预计手术完全恢复。 </li><li>手术后将动物放在加热垫（避免体温过低）和吸收性软床上。 </li><li>提供口服或肠胃外液体的补液，直到它能够恢复正常进食（24小时内）。手术后立即口服约0.6ml生理盐水溶液以限制脱水效果。 </li><li>按照IACUC批准的动物保健和使用协议中所述提供止痛。皮下注射5 mg / kg卡洛芬用于疼痛管理。 </li><li>定期监测切口是否肿胀，渗出，疼痛或开裂。 </li></ol> 6. 通过磁共振成像验证<ol><li>对高场小动物MRI扫描仪进行体内 MRI实验。 </li><li>用3％异氟烷在氧气中诱导麻醉，并将动物与右肾在等中心放置在MRI扫描仪中吃射频线圈。在整个成像过程中通过鼻锥连续地给动物1-2％的异氟烷​​麻醉。 注意:对于图1所示的体内 MRI图像，使用大鼠体积线圈（内径= 72mm）。 </li><li>使用动物监测和控制系统来维持每只动物的呼吸速率（40-60次呼吸/分钟）和核心体温（35±1℃）。 </li><li>使用多层，多回波自旋回波MRI采集获得感染和对照肾脏的高分辨率，轴向T2加权图像。典型的MRI采集参数是重复时间= 5000msec，回波时间= 40msec，切片厚度= 2.0mm，平面内空间分辨率=200μm，3个信号平均值，并且采集时间为8分钟。 </li></ol>

<ol><li>Saliba, W., Barnett-Griness, O., Rennert, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+association+between+obesity+and+urinary+tract+infection%5BTitle%5D)+AND+%22Eur+J+Intern+Med%22%5BJournal%5D)">The association between obesity and urinary tract infection.</a> Eur J Intern Med. 24 (2), 27-31 (2012).</li>
<li>Semins, M., Shore, A., Makary, M., Weiner, J., Matlaga, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+impact+of+obesity+on+urinary+tract+infection+risk%5BTitle%5D)+AND+%22Urology%22%5BJournal%5D)">The impact of obesity on urinary tract infection risk.</a> Urology. 79 (2), 266-269 (2011).</li>
<li>Zilberberg, M., Shorr, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Secular+trends+in+gram-negative+resistance+among+urinary+tract+infection+hospitalizations+in+the+United+States%2C+2000-2009%5BTitle%5D)+AND+%22Infect+Control+Hosp+Epidemiol%22%5BJournal%5D)">Secular trends in gram-negative resistance among urinary tract infection hospitalizations in the United States, 2000-2009.</a> Infect Control Hosp Epidemiol. 34 (9), 940-946 (2013).</li>
<li>Whiting, D., Guariguata, L., Weil, C., Shaw, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((IDF+diabetes+atlas%3A+global+estimates+of+the+prevalence+of+diabetes+for+2011+and+2030%5BTitle%5D)+AND+%22Diabetes+Res+Clin+Pract%22%5BJournal%5D)">IDF diabetes atlas: global estimates of the prevalence of diabetes for 2011 and 2030.</a> Diabetes Res Clin Pract. 94 (3), 311-321 (2011).</li>
<li>Wild, S., Roglic, G., Green, A., Sicree, R., King, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Global+prevalence+of+diabetes%3A+estimates+for+the+year+2000+and+projections+for+2030%5BTitle%5D)+AND+%22Diabetes+Care%22%5BJournal%5D)">Global prevalence of diabetes: estimates for the year 2000 and projections for 2030.</a> Diabetes Care. 27 (5), 1047-1053 (2004).</li>
<li>Ma, D., Gulani, V., Seiberlich, N., Liu, K., Sunshine, J., Duerk, J., et al. <a target="_blank" href="https://www.nature.com/nature/journal/v495/n7440/full/nature11971.html">Magnetic resonance fingerprinting.</a> Nature. 495 (7440), 187-192 (2013).</li>
<li>Lu, L., Sedor, J., Gulani, V., Schelling, J., O'Brien, A., Flask, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Use+of+diffusion+tensor+MRI+to+identify+early+changes+in+diabetic+nephropathy%5BTitle%5D)+AND+%22Am+J+Nephrol%22%5BJournal%5D)">Use of diffusion tensor MRI to identify early changes in diabetic nephropathy.</a> Am J Nephrol. 34 (5), 476-482 (2011).</li>
<li>Rosen, D., Hooton, T., Stamm, W., Humphrey, P., Hultgren, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Detection+of+intracellular+bacterial+communities+in+human+urinary+tract+infection%5BTitle%5D)+AND+%22PLoS+Med%22%5BJournal%5D)">Detection of intracellular bacterial communities in human urinary tract infection.</a> PLoS Med. 4 (12), e329(2007).</li>
<li>Torine, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Urinary+tract+infection%3A+diabetic+women%27s+strategies+for+prevention%5BTitle%5D)+AND+%22Br+J+Nurs%22%5BJournal%5D)">Urinary tract infection: diabetic women's strategies for prevention.</a> Br J Nurs. 20 (13), 791-792 (2011).</li>
<li>Rosen, D., Hung, C., Kline, K., Hultgren, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Streptozocin-induced+diabetic+mouse+model+of+urinary+tract+infection%5BTitle%5D)+AND+%22Infect+Immun%22%5BJournal%5D)">Streptozocin-induced diabetic mouse model of urinary tract infection.</a> Infect Immun. 76 (9), 4290-4298 (2008).</li>
<li>Larsson, P., Kaijser, B., Mattsby-Baltzer, I., Olling, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+experimental+model+for+ascending+acute+pyelonephritis+caused+by+Escherichia+coli+or+proteus+in+rats%5BTitle%5D)+AND+%22J+Clin+Pathol%22%5BJournal%5D)">An experimental model for ascending acute pyelonephritis caused by Escherichia coli or proteus in rats.</a> J Clin Pathol. 33 (4), 408-412 (1980).</li>
<li>Gupta, R., Ganguly, N., Ahuja, V., Joshi, K., Sharma, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+ascending+non-obstructive+model+for+chronic+pyelonephritis+in+BALB%2Fc+mice%5BTitle%5D)+AND+%22J.+Med.+Microbiol%22%5BJournal%5D)">An ascending non-obstructive model for chronic pyelonephritis in BALB/c mice.</a> J. Med. Microbiol. 43 (1), 33-36 (1995).</li>
<li>Fernandes, P., Shipkowitz, N., Bower, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Murine+models+for+studying+the+pathogenesis+and+treatment+of+pyelonephritis%5BTitle%5D)+AND+%22Adv.+Exp.+Med.+Biol%22%5BJournal%5D)">Murine models for studying the pathogenesis and treatment of pyelonephritis.</a> Adv. Exp. Med. Biol. 224, 35-51 (1987).</li>
<li>Kaye, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+effect+of+water+diuresis+on+spread+of+bacteria+through+the+urinary+tract%5BTitle%5D)+AND+%22J.+Infect.+Dis%22%5BJournal%5D)">The effect of water diuresis on spread of bacteria through the urinary tract.</a> J. Infect. Dis. 124 (3), 297-305 (1971).</li>
<li>Fierer, J., Tainer, L., Braude, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Bacteremia+in+the+pathogenesis+of+retrograde+E.+coli+pyelonephritis+in+the+rat%5BTitle%5D)+AND+%22Am.+J.+Pathol%22%5BJournal%5D)">Bacteremia in the pathogenesis of retrograde E. coli pyelonephritis in the rat.</a> Am. J. Pathol. 64 (2), 443-456 (1971).</li>
<li>Nickel, J., Olson, M., Costerton, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rat+model+of+experimental+bacterial+prostatitis%5BTitle%5D)+AND+%22Infection%22%5BJournal%5D)">Rat model of experimental bacterial prostatitis.</a> Infection. 19 (3), S126-S130 (1991).</li>
<li>Hagberg, L., Engberg, I., Freter, R., Olling, S., Eden, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ascending%2C+unobstructed+urinary+tract+infection+in+mice+caused+by+pyelonephritogenic+Escherichia+coli+of+human+origin%5BTitle%5D)+AND+%22Am+Soc+Microbiol%22%5BJournal%5D)">Ascending, unobstructed urinary tract infection in mice caused by pyelonephritogenic Escherichia coli of human origin.</a> Am Soc Microbiol. 40 (1), 273-283 (1983).</li>
<li>Kurosaka, Y., Ishida, Y., Yamamura, E., Takase, H., Otani, T., Kumon, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+non-surgical+rat+model+of+foreign+body-associated+urinary+tract+infection+with+Pseudomonas+aeruginosa%5BTitle%5D)+AND+%22Microbiol.+Immunol%22%5BJournal%5D)">A non-surgical rat model of foreign body-associated urinary tract infection with Pseudomonas aeruginosa.</a> Microbiol. Immunol. 45 (1), 9-15 (2001).</li>
<li>Anderson, B., Jackson, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pyelitis%2C+an+important+factor+in+the+pathogenesis+of+retrograde+pyelonephritis%5BTitle%5D)+AND+%22J+Exp+Med%22%5BJournal%5D)">Pyelitis, an important factor in the pathogenesis of retrograde pyelonephritis.</a> J Exp Med. 114 (3), 375-384 (1961).</li>
<li>Anderson, J. <a target="_blank" href="http://journals.sagepub.com/doi/pdf/10.1177/003591576205500601">Vesico-ureteric reflux.</a> J R Soc Med. 55 (6), 419-426 (1962).</li>
</ol>

The authors have no other disclosures.

啮齿动物（即小鼠和大鼠）升高的急性肾盂肾炎可以通过经尿道导尿术产生。 16，17，18经尿道这种感染的方法是有利的，因为它是非侵入性的和模拟上行感染的人类病理生理学。 17，18，19，20然而，这种方法也可以遭受不?...

啮齿动物模型已被用于研究许多人类疾病表现，包括肾盂肾炎和尿路感染（UTI）。 UTI是一个全球性的健康问题，可以影响所有年龄段的儿童，男性和女性。 1，2，3尿路感染的初始表现形式包括膀胱炎，如果感染沿输尿管上升，肾脏感染（肾盂肾炎）可以遵循。与此同时，全球糖尿病患病率接近四亿人。 4,5重要的是，尿路感染发病率可能高达谁是病人肥胖或有2型糖尿病，导致复发性尿路感染感染（入替），败血症，从肾盂肾炎肾纤维化和膀胱功能障碍的风险增加4倍以上升高。 6，7，8鼠类模型在研究UTI方面很重要，因为目前的抗生素疗法仅在UTI患者的一个子集中产生持续的预防性应答。为了改善临床UTI护理，关键步骤是了解rUTI及其从急性感染到炎症到纤维化的病理生理过程的机制以及2型糖尿病的影响。 改进动物模型的目标是开发可以更准确地评估疾病进展和治疗干预措施的技术。已经采用几种不同的方法在大鼠和/或小鼠中诱导肾盂肾炎，以研究肾损伤的病理生理学，抗生素治疗的效果以及UTI的自然过程的其他方面。建立逆行性UTI的常见方法是经尿道导尿。 10，11，12，13 &lt;/ sup&gt;该方法通过尿道将细菌引入麻醉动物的膀胱。虽然这种技术密切模拟人类肾盂肾炎，但由于多种因素，肾盂肾炎感染的实际发生率和大小可能是非常可变的，包括在接种期间或之后紧随其后的输尿管反流自发或排尿不足。 11其结果是，在诱导上行肾盂肾炎的感染可以限制这种模型来研究肾脏感染以及治疗策略的效用的实验变化。 该报告描述了一种手术肾盂肾炎大鼠模型，其中将大肠杆菌直接注射入大鼠肾脏。尽管这种大鼠模型是侵入性的，但是可以有效地控制输送到肾脏的大肠杆菌的数量，从而能够强壮的肾脏感染和炎症。 14在这个过程中，我们还描述如何通过体内磁共振成像（MRI）纵向监测这些诱导的肾脏感染。

<table><tbody><tr><td>Absorbing Pad</td><td>Fisher</td><td>14-127-47</td><td></td></tr><tr><td>Sterile Cotton Gauze Pad</td><td>Fisher</td><td>22-415-469</td><td></td></tr><tr><td>Latex Surgical Gloves</td><td>Henry Schein Animal Health</td><td>21540</td><td></td></tr><tr><td>Curved Mayo Scissors</td><td>Fisher</td><td>S17341</td><td></td></tr><tr><td>Straight Blunt Foreceps</td><td>Fisher</td><td>08-895</td><td></td></tr><tr><td>Scalpel Handle&amp;nbsp;</td><td>Fisher</td><td>08-913-5</td><td></td></tr><tr><td>Sterile Scalpel Blades</td><td>Fisher</td><td>53220</td><td></td></tr><tr><td>1 ml Luer-Lok Syringe</td><td>BD Biosciences</td><td>309628</td><td>For bacterial injections</td></tr><tr><td>20 ml Luer-Lok Syringe&amp;nbsp;</td><td>BD Biosciences</td><td>301031</td><td>For saline wash</td></tr><tr><td>Hemostat</td><td>Seneca Medical</td><td>240267</td><td></td></tr><tr><td>23 G 3/4 in. Needle&amp;nbsp;</td><td>BD Biosciences</td><td>305143</td><td></td></tr><tr><td>30 G 1 in. Needle&amp;nbsp;</td><td>BD Biosciences</td><td>305128</td><td></td></tr><tr><td>U-100 Insulin Syringe</td><td>Exel International&amp;nbsp;</td><td>25846</td><td>For medication injections</td></tr><tr><td>Isoflurane</td><td>Henry Schein Animal Health</td><td>050033</td><td></td></tr><tr><td>Xylazine</td><td>Henry Schein Animal Health</td><td>33197</td><td>Inject IP</td></tr><tr><td>Ketamine</td><td>Patterson Vetrinary&amp;nbsp;</td><td>07-881-9413</td><td>Inject IP</td></tr><tr><td>Yohimbine (Atipamezole)</td><td>Patterson Vetrinary&amp;nbsp;</td><td>07-867-7097</td><td>Inject IP after surgery</td></tr><tr><td>Bupivacaine (Marcaine)</td><td>Patterson Vetrinary&amp;nbsp;</td><td>07-890-4584</td><td>Inject SQ at site of incision&amp;nbsp;</td></tr><tr><td>4-0 Chromic Gut Suture</td><td>Ethicon Inc.</td><td>U203H</td><td></td></tr><tr><td>4-0 Braided Vicryl Suture</td><td>Ethicon Inc.</td><td>J304H</td><td></td></tr><tr><td>1 ml SubQ Syringe</td><td>BD Biosciences</td><td>309597</td><td></td></tr><tr><td>&lt;em&gt;E. coli&lt;/em&gt;&amp;nbsp; UTI89 or CFT073</td><td>ATCC</td><td>700928</td><td></td></tr><tr><td>Surgicel Absorbable Hemostat</td><td>Ethicon Inc.</td><td>ETH1951CS&amp;nbsp;</td><td></td></tr><tr><td>Biospec 9.4T MRI&amp;nbsp;</td><td>Bruker&amp;nbsp;</td><td>94/20 USR</td><td></td></tr></tbody></table>

intrarenal injection of escherichia coli in a rat model of pyelonephritis

Pyelonephritis is a bacterial infection of the kidney and is most commonly caused by Escherichia coli. Recurrent infections can cause significant renal inflammation and fibrosis ultimately resulting in declining kidney function. Before improved clinical management and prevention of pyelonephritis can be instituted, a reliable animal model must be established in order to study the mechanisms of progression, recurrence, and therapeutic efficacy. The transurethral infection model closely mimics human pyelonephritis but exhibits considerable variation due to its reliance on urethral reflux to transport the bacteria to the kidney. Herein, a detailed surgical protocol for performing bacterial injections into the rat renal pelvis is provided and confirmed by non-invasive Magnetic Resonance Imaging (MRI). Using this protocol, animals receive direct exposure to a desired concentration of E. coli bacteria and can fully recover from the surgical procedure with adequate post-operative care. This facilitates subsequent longitudinal MRI assessments of the experimental animal models for comparison with saline (sham) controls. Using this direct delivery approach, the severity of infection is controllable and applicable for mechanistic studies of progression as well as development of novel treatment strategies.

医学成像技术提供了非侵入性评估UTI和治疗功效的机会。因此，MRI用于验证注射1-2×10 7 UTI89 大肠杆菌后的急性感染诱导，并可视化手术前后肾脏的变化。 图1a-b显示肾脏感染逐渐增加的区域（黄色箭头）。在感染后第1天和第4天为每只动物获得的MRI图像有助于表征急性肾盂肾炎感染的生长。 <img alt="?...

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Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

This manuscript describes a rat surgical model of pyelonephritis using direct intra-renal infection by Escherichia coli into the renal pelvis. The experimental procedure can be utilized to study the pathogenesis of pyelonephritis as well as the associated inflammation and fibrosis.

肾内注射<em&gt;大肠杆菌</em&gt;在肾性肾炎大鼠模型中

Pyelonephritis is a bacterial infection of the kidney and is most commonly caused by Escherichia coli. Recurrent infections can cause significant renal ...

intrarenal-injection-escherichia-coli-rat-model

Research

JoVE Journal

Medicine

Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

8.0K 次观看.  Case Western Reserve University. This manuscript describes a rat surgical model of pyelonephritis using direct intra-renal infection by Escherichia coli into the renal pelvis. The experimental procedure can be utilized to study the pathogenesis of pyelonephritis as well as the associated inflammation and fibrosis.

视频: Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

Transurethral Induction of Mouse Urinary Tract Infection

Uropathogenic bacterial strains of interest are grown on agar. Generally, uropathogenic E. coli (UPEC) and other strains can be grown overnight on Luria-Bertani (LB) agar at 37&deg;C in ambient air. UPEC strains grow as yellowish-white translucent colonies on LB agar. Following confirmation of appropriate colony morphology, single colonies are then picked to be cultured in broth. LB broth can be used for most uropathogenic bacterial strains. Two serial, overnight LB broth cultures can be employed to enhance expression of type I pili, a well-defined virulence factor for uropathogenic bacteria. Broth cultures are diluted to the desired concentration in phosphate buffered saline (PBS). Eight to 12 week old female mice are placed under isoflurane anesthesia and transurethrally inoculated with bacteria using polyethylene tubing-covered 30 gauge syringes. Typical inocula, which must be empirically determined for each bacterial/mouse strain combination, are 106 to 108 cfu per mouse in 10 to 50 microliters of PBS. After the desired infection period (one day to several weeks), urine samples and the bladder and both kidneys are harvested. Each organ is minced, placed in PBS, and homogenized in a Blue Bullet homogenizer. Urine and tissue homogenates are serially diluted in PBS and cultured on appropriate agar. The following day, colony forming units are counted.

This video will demonstrate methods to transurethrally induce mouse urinary tract infections and quantify the extent of resulting infections.

经尿道诱导小鼠尿路感染

Uropathogenic bacterial strains of interest are grown on agar. Generally, uropathogenic E. coli (UPEC) and other strains can be grown overnight on ...

科研

免疫与感染

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Investigation of the interactions between animal host and bacterial pathogen is only meaningful if the infection model employed replicates the principal features of the natural infection. This protocol describes procedures for the establishment and evaluation of systemic infection due to neuropathogenic Escherichia coli K1 in the neonatal rat. Colonization of the gastrointestinal tract leads to dissemination of the pathogen along the gut-lymph-blood-brain course of infection and the model displays strong age dependency. A strain of E. coli O18:K1 with enhanced virulence for the neonatal rat produces exceptionally high rates of colonization, translocation to the blood compartment and invasion of the meninges following transit through the choroid plexus. As in the human host, penetration of the central nervous system is accompanied by local inflammation and an invariably lethal outcome. The model is of proven utility for studies of the mechanism of pathogenesis, for evaluation of therapeutic interventions and for assessment of bacterial virulence.

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Here, a procedure is described for the establishment of systemic infection in the neonatal rat with cultures of Escherichia coli K1. This non-invasive procedure permits colonization of the gastrointestinal tract, translocation of the pathogen to the systemic circulation, and invasion of the central nervous system at the choroid plexus.

Neuropathogenic的非侵入型<em&gt;大肠杆菌</em&gt;感染乳鼠

Investigation of the interactions between animal host and bacterial pathogen is only meaningful if the infection model employed replicates the principal ...

Urinary Tract Infection in a Small Animal Model: Transurethral Catheterization of Male and Female Mice

Urinary tract infections (UTI) are extremely common worldwide, incurring significant morbidity and healthcare-associated expenses. Small animal models, which accurately reflect disease establishment and progression, permit dissection of host-pathogen interactions and generation of immunity to infection. In mice, intravesical instillation of uropathogenic E. coli, the causative agent in more than 85% of community acquired UTI, recapitulates many of the stages of infection observed in humans. Until recently, however, UTI could only be modeled in female animals. This limitation has hindered the study of sex-related differences in UTI, as well as other bladder pathologies, such as cancer. Here, we describe a method to instill male mice that allows direct comparison between female and male animals and provide a detailed protocol to assess bladder tissue by flow cytometry as a means to better understand host responses to infection. Together, these approaches will aid in the identification of host factors that contribute to sex biases observed in UTI and other bladder-associated diseases.

The established model of transurethral catheterization of mice allows the study of bladder pathologies, including urinary tract infection, but can only be performed in females. A new model of male transurethral instillation, presented here, will enable research in an area marked by strong clinical and epidemiological differences between the sexes.

小动物模型尿路感染的实验研究: 男性和女性的经尿道插管

Urinary tract infections (UTI) are extremely common worldwide, incurring significant morbidity and healthcare-associated expenses. Small animal models, ...

Standardized Colon Ascendens Stent Peritonitis in Rats - a Simple, Feasible Animal Model to Induce Septic Acute Kidney Injury

AKI in septic patients is associated with increased mortality and poor outcome despite major efforts to refine the understanding of its pathophysiology. Here, an in vivo model is presented that combines a standardized septic focus to induce AKI and an intensive care (ICU) setup to provide an advanced hemodynamic monitoring and therapy comparable in human sepsis. Sepsis is induced by standardized colon ascendens stent peritonitis (sCASP). AKI is investigated functionally by measurement of blood and urine samples as well as histologically by evaluation of histopathological scores. Furthermore, the advanced hemodynamic monitoring and the possibility of repetitive blood gas sampling enable a differentiated analysis of severity of induced sepsis.
The sCASP method is a standardized, reliable and reproducible method to induce septic AKI. The intensive care setup, continuous hemodynamic and gas exchange monitoring, low mortality rate as well as the opportunity of detailed analyses of kidney function and impairments are advantages of this setup. Therefore, the described method may serve as a new standard for experimental investigations of septic AKI.

<meta charset="utf8"></head><body>大鼠标准化结肠升行支架腹膜炎 - 一种诱导脓毒性急性肾损伤的简单、可行的动物模型

Acute kidney injury (AKI) is a severe complication in critically ill patients and is related with an increased mortality. Here, we present a reliable and reproducible in vivo model to mimic AKI under inflammatory conditions that might contribute towards understanding the pathogenesis of septic AKI.

大鼠标准化结肠升行支架腹膜炎 - 一种诱导脓毒性急性肾损伤的简单、可行的动物模型

AKI in septic patients is associated with increased mortality and poor outcome despite major efforts to refine the understanding of its pathophysiology. ...

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of UTI in male and female mice have illustrated the procedures for bacterial inoculation and enumeration in urine and tissues. During an initial bladder infection in C57BL/6 mice, UPEC establish latent reservoirs inside bladder epithelial cells that persist following clearance of UPEC bacteriuria. This model builds on these studies to examine rUTI caused by the emergence of UPEC from within latent bladder reservoirs. The urogenital bacterium Gardnerella vaginalis is used as the trigger of rUTI in this model because it is frequently present in the urogenital tracts of women, especially in the context of vaginal dysbiosis that has been associated with UTI. In addition, a method for in situ bladder fixation followed by scanning electron microscopy (SEM) analysis of bladder tissue is also described, with potential application to other studies involving the bladder.

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

A mouse model of uropathogenic E. coli (UPEC) transurethral inoculation to establish latent intracellular bladder reservoirs and subsequent bladder exposure to G. vaginalis to induce recurrent UPEC UTI is demonstrated. Also demonstrated are the enumeration of bacteria, urine cytology, and in situ bladder fixation and processing for scanning electron microscopy.

由加德纳菌阴道膀胱暴露在小鼠中引发的复发性大肠杆菌尿路感染

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of ...

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

Acute kidney injury (AKI) is associated with higher risk for morbidity and mortality post-operatively. Ischemia-reperfusion injury (IRI) is the most common cause of AKI. To mimic this clinical scenario, this study presents a highly reproducible large animal model of renal IRI in swine using temporary percutaneous bilateral balloon-catheter occlusion of the renal arteries. The renal arteries are occluded for 60 min by introducing the balloon-catheters through the femoral and carotid artery and advancing them into the proximal portion of the arteries. Iodinated contrast is injected in the aorta to assess any opacification of the kidney vessels and confirm the success of the artery occlusion. This is furtherly confirmed by the flattening of the pulse waveform at the tip of the balloon catheters. The balloons are deflated and removed after 60 min of bilateral renal artery occlusion, and the animals are allowed to recover for 24 h. At the end of the study, plasma creatinine and blood urea nitrogen significantly increase, while eGFR and urine output significantly decrease. The need for iodinated contrast is minimal and does not affect renal function. Bilateral renal artery occlusion better mimics the clinical scenario of perioperative renal hypoperfusion, and the percutaneous approach minimizes the impact of the inflammatory response and the risk of infection seen with an open approach, such as a laparotomy. The ability to create and reproduce this clinically relevant swine model eases the clinical translation to humans.

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.

临时双性肾动脉闭塞的急性肾损伤大型动物模型

Acute kidney injury (AKI) is associated with higher risk for morbidity and mortality post-operatively. Ischemia-reperfusion injury (IRI) is the most ...

医学

Hydrodynamic Renal Pelvis Injection: A Technique to Directly Deliver DNA into Kidney Cells of Murine Model

Source: Woodard, L. E. et al. <a href="https://www.jove.com/video/56324" target="_blank">Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney.</a> J. Vis. Exp. (2018)
In this video, we present the protocol for hydrodynamic injection of plasmids into the renal pelvis of the mouse kidney for nucleic acid transfection. This technique allows organ-specific delivery of plasmids by utilizing the principle of high hydrostatic pressure, which forcefully drives the plasmids into the specific site without causing tissue injury.

流体动力学肾盂注射液:一种将 DNA 直接递送到小鼠肾细胞模型的技术

Source: Woodard, L. E. et al. Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney. J. Vis. Exp. (2018)
In this video, ...

JoVE Encyclopedia of Experiments

癌症研究

Urinary Tract Cancer

In vivo studies

Transurethral Catheterization and Bacterial Inoculation to Generate a Urinary Tract Infection Model in Male Mice

Source: Zychlinsky Scharff, A., et al. <a href="https://app.jove.com/v/54432">Urinary Tract Infection in a Small Animal Model: Transurethral Catheterization of Male and Female Mice.</a> J. Vis. Exp. (2017).
This video demonstrates the establishment of uropathogenic Escherichia coli infection in a female mouse bladder through transurethral catheter instillation. The injected bacteria successfully establish a urinary tract infection within the bladder, providing a valuable platform for studying healthcare-associated diseases related to the urinary tract.

经尿道导管插入术和细菌接种在雄性小鼠中生成尿路感染模型

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

Immunology

Immunopathology

Host-Pathogen Interaction Models

Establishment of a Urinary Tract Infection Model in Female Mice

Source: Zychlinsky Scharff, A. et al., <a href="https://app.jove.com/v/54432">Urinary Tract Infection in a Small Animal Model: Transurethral Catheterization of Male and Female Mice.</a>&#160;J. Vis. Exp. (2017)
This video demonstrates the establishment of uropathogenic Escherichia coli infection in a female mouse bladder through transurethral catheter instillation. The injected bacteria successfully establish a urinary tract infection within the bladder, providing a valuable platform for studying healthcare-associated diseases related to the urinary tract.

肾内注射

摘要

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经尿道诱导小鼠尿路感染

Neuropathogenic的非侵入型

小动物模型尿路感染的实验研究: 男性和女性的经尿道插管

大鼠标准化结肠升行支架腹膜炎 - 一种诱导脓毒性急性肾损伤的简单、可行的动物模型

大鼠标准化结肠升行支架腹膜炎 - 一种诱导脓毒性急性肾损伤的简单、可行的动物模型

由加德纳菌阴道膀胱暴露在小鼠中引发的复发性大肠杆菌尿路感染

临时双性肾动脉闭塞的急性肾损伤大型动物模型

流体动力学肾盂注射液:一种将 DNA 直接递送到小鼠肾细胞模型的技术

经尿道导管插入术和细菌接种在雄性小鼠中生成尿路感染模型

雌性小鼠尿路感染模型的建立