Name: urinary tract infection in a small animal model: transurethral catheterization of male and female mice
Uploaded: 2017-12-01T13:00:00.000+00:00
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Description: Watch this Scientific Journal Video about Urinary Tract Infection in a Small Animal Model: Transurethral Catheterization of Male and Female Mice at JoVE.com

我们感谢马修博士在本议定书和项目的发展过程中对树突免疫的手稿和实验室的批判性阅读。这项工作得到了欧洲联盟第七框架方案居里夫人行动 (PCIG11-GA-2012-3221170 和免疫肿瘤学 LabEx (马伊) 的资助。

根据éthique en expérimentation animale 巴黎中心和南方 (动物实验伦理学委员会) 批准的2012-0024 号议定书, 在应用欧洲指令2010/63 时进行了老鼠实验。欧盟.
1. 导管的制备
<ol><li>为每组被感染的小鼠准备一个儿科静脉插管。使用内置弹簧机构, 剥离其针的每个套管, 按照制造商的指示。丢弃针头, 只保留塑料静脉导管。</li>
	<li>在层流罩内消毒导管, 用于一个紫外线 (紫外线) 循环, 通常为 25-30 分钟。 注: 导管可用于多只鼠标, 但应在实验组、性别或细菌菌株之间使用新的导管。</li>
</ol>2. 为感染而制备肾炎的大肠杆菌
<ol><li>至少在感染前两天, 使用无菌接种回路, 条纹 UPEC 从一个冷冻细菌库存到 Luria 肉汤 (LB) 琼脂板, 含有抗生素, 如果适当的。在晚上37° c 孵育。板材可储存在4° c, 最多一周。</li>
	<li>在一个100毫升无菌锥形烧瓶, 接种10毫升磅, 其中含有抗生素, 如果合适的, 一个单一的殖民地从 LB 板和孵化站在37° c 过夜 (约 16-18 小时)。 注意: 站立区域性允许类型1毛的表达式, 这是感染23所必需的。文化的颤抖将有较低的效率毛表达, 因此, 不一致的感染率。</li>
	<li>测量过夜培养的光学密度 (OD)600 , 并使用预先确定的生长曲线计算每毫升细菌的数量。举例来说, 对于 UPEC 应变 UTI89, od600 = 0.35 等于每毫升 2 x 108 CFU, 因此, 具有 od600 = 2.4 的隔夜区域性相当于 13.7 x 108 CFU 每毫升. 根据经验确定OD600和可存活的细菌与每一个新的菌株使用的感染。</li>
	<li>
通过考虑被感染动物的数量来确定所需的细菌数量, 并在50μ l PBS 中每只老鼠接收大约 107菌落形成单位 (CFU)。在计算中包括130µl 死亡体积的导管和注射器的核心和100µl 需要确定接种。
	<ol>
<li>旋转的细菌悬浮在一个桌面离心在 1.7万 x g 为1分钟和重的结果细菌颗粒在 2 x 108 CFU 每毫升在 PBS。连续地稀释这个悬浮和板材的分在 LB 琼脂, 与抗生素, 如果适当, 确定确切的接种为每传染。</li>
	</ol>
</li>
	<li>将细菌接种成1毫升的注射器, 并将导管连接到注射器的末端。在开始灌输前, 轻敲注射器以除去任何空气, 并压低柱塞以填满导管内的死亡空气空间。</li>
</ol>3. 小鼠的制备
<ol><li>麻醉小鼠通过腹腔注射100毫克/千克氯胺酮和5毫克/千克嗪。可提供补充热量。</li>
	<li>确保每只老鼠都通过挤压脚中压来充分镇静。小鼠完全镇静, 当他们没有反应, 并要求 3-5 分钟达到这个状态。</li>
	<li>将小鼠仰卧, 应用中压到下腹, 排空尿液中的膀胱。全膀胱感觉像一个豌豆下的皮肤之间的回肠波峰。 注: 吸入异氟醚已被用于 catheterize 雌性小鼠15,22, 但是, 该方法尚未对雄性小鼠是否已被吸入异氟醚充分麻醉进行了测试。</li>
</ol>4. 经尿道灌注的雌性小鼠
<ol><li>使用非显性的手, 将拇指放在鼠标的腹部和手指上, 并在相反的方向施加柔和的压力, 以使鼠标牢牢地固定到位。</li>
	<li>
将导管的尖端垂直于老鼠的尿道口。用轻柔的压力, 将导管滑入尿道, 直到轮毂碰到尿道口, 同时降低注射器, 使其与工作表面平行。导管不需要润滑。不要将导管推入尿道。导管应平滑滑入尿道, 几乎无阻力。
	<ol>
<li>在灌输接种 (步骤 4.3) 之前, 一旦导管到位, 就用手指从已经在鼠标腹部的非显性手轻轻地将腹部皮肤拉向鼠标的头部 (步骤 4.1)。如果导管在尿道内, 构成尿道口的组织不会移动, 而如果导管不正确放置在阴道内, 则组织会向上和远离导管。这种快速测试应该在每只老鼠身上进行。</li>
	</ol>
</li>
	<li>当导管的中心与尿道口相遇时, 慢慢免除50µl 的细菌接种。缓慢的灌注率将膀胱的回流降到肾脏。慢慢地取出导管以防止渗漏, 超过数5。把老鼠放在笼子里放在仰卧位。</li>
</ol>5. 经尿道灌注的雄性小鼠
<ol><li>放置两个拇指钳 cranially 和尾到鼠标的外生殖器。收回包皮以充分暴露头头的阳具。一旦阳具外部定位, 松开拇指钳。</li>
	<li>重新定位钳, 以稳定突出的生殖器垂直的动物, 轻轻地拿着器官, 但紧。可视化尿道道, 并小心地将导管插入器官尖端的小开口处。将导管轻轻地引导到阳具中, 朝向老鼠的身体, 用镊子保持温和的张力。导管不需要润滑。不要强迫导管进入生殖器;导管应平滑滑入尿道, 指示正确放置。</li>
	<li>一旦导管的中心与生殖器的尖端, 非常缓慢地免除50µl 的接种, 同时保持的位置的小弟弟。根据预先确定的灌输质量分数 (如图 2所示), 此时可以注意到灌输的质量。</li>
	<li>慢慢缩回导管, 超过数 5, 以防止接种的渗漏。把老鼠放在笼子里放在仰卧位。动物应开始恢复 30-45 分钟后的麻醉管理。</li>
</ol>6. 感染器官中 CFU 的测定
<ol><li>在预定的时间点, 牺牲小鼠使用批准的标准操作程序 (例如, 颈椎脱位或二氧化碳过量)。用70% 乙醇彻底滋润腹部, 以减少毛皮污染。</li>
	<li>用剪刀和菌将至少2厘米的小鼠腹部的下三分之一切开, 除去膀胱和其他所需器官, 包括但不限于肾脏、睾丸、精囊或包皮腺体, 到5毫升含有1毫升无菌 PBS 的聚丙烯快盖管。把所有的样品放在冰上。</li>
	<li>用手持式均质器质器官, 直到几乎没有固体组织残留, 大约 15-60 秒取决于器官。脂肪不质好, 容易辨认为闪亮的米色白色组织。在同质化之前或之后丢弃脂肪。在每个实验或器官组之间, 用乙醇清洗匀浆器。 注: 通常使用手持质或珠磨质。虽然所有这些系统都产生了类似的结果, 通过直接比较细菌 CFU 在24小时后感染, 质宿主组织的最佳条件, 同时保留细菌的生存能力, 应在经验前确定例行执行此协议。</li>
	<li>执行均匀化器官悬浮的串联稀释。平板稀释在 LB 琼脂板上, 适当时含有抗生素, 在37° c 时孵育过夜 (约15小时)。UPEC 菌株有非常快的倍倍, 因此, 应采取谨慎不孵化琼脂板太长, 因为这将阻碍能力计数个别殖民地。</li>
</ol>7. 膀胱组织的流动细胞分析
<ol><li>在 PBS 的100µg/毫升, 准备34单位/毫升和 dnasei 的消化缓冲液中含有胶原酶。继续冰每只动物准备一个15毫升的锥管进行分析, 每管分1毫升消化缓冲液。</li>
	<li>在预定的 timepoints, 使用批准的标准操作程序 (例如,颈椎脱位或二氧化碳过量) 牺牲小鼠。用70% 乙醇彻底滋润腹部, 以减少毛皮污染。</li>
	<li>
用剪刀将至少2厘米的小鼠腹部的下三分之一切开, 并取出整个膀胱。用剪刀切碎, 将膀胱切成 2-3 mm2大小的碎片, 通常约8件。
	<ol>
<li>每管含消化缓冲液加一个切碎的膀胱。摇动以将切碎的膀胱组织冲洗到消化缓冲液中。继续冰, 直到所有膀胱被解剖和剁碎。</li>
	</ol>
</li>
	<li>在37° c 下孵育 60-75 分钟的碎膀胱, 用手剧烈摇动5秒, 每15分钟。当组织有玻璃透明的外观, 类似于湿纸巾, 消化是完整的。长时间的消化将从细胞中去除表面蛋白, 以供鉴别, 应避免。</li>
	<li>通过加入 2-3 毫升的冰-冷流式细胞仪缓冲液 (PBS 含有2% 胎牛血清 (FBS) 和0.2 毫米 EDTA) 使消化酶钝化, 并轻轻混合。把管子放在冰上。</li>
	<li>为了确保单个细胞悬浮, 并去除任何结缔组织, 通过100µm 过滤器放置在一个15毫升锥形管管的内容。用注射器柱塞的末端轻轻按压任何剩余的组织。用额外的2毫升流式细胞仪缓冲清洗过滤器。把样品放在冰上。</li>
	<li>用离心法洗涤样品7分钟, 在 200 x g, 4 ° c。重颗粒在100µL 流式细胞仪缓冲包含 Fc 块, 稀释到5µg/毫升和转移到一个96井圆底板。10-15 分钟后, 在每个样本中添加100µL 所需的抗体鸡尾酒。在冰上孵育样品 30-45 分钟, 免受光照。 注: fc 块是一种试剂, 用于防止抗体 fc 部分与表达 fc 受体的细胞结合。其目的是尽量减少非特异性抗体结合。使用的抗体的测定将取决于在实验中测试的假说, 并应选择从文献的输入。在图 3中, 为了测量整体免疫细胞浸润, 采用了一种 anti-CD45 抗体, 它是一种泛免疫细胞蛋白。</li>
	<li>用离心法洗涤样品7分钟, 在 200 x g, 4 ° c。重细胞颗粒在200µL 流式细胞仪缓冲和通过一个40µm 细胞过滤器上的5毫升聚苯乙烯管, 刚刚获得的流量胞内液。</li>
	<li>收集整个样品流式细胞仪。一个良好的消化将产生 20万-40万细胞, 8000-1万 CD45+免疫细胞从天真的小鼠膀胱, 而感染器官将包含更多的细胞16 (图 3)。 注: 为流细胞分析准备的样品不能用于评价 CFU。不建议将膀胱分成两部分, 因为没有证据证明 UPEC 定植或免疫细胞浸润在整个组织中都是均匀的。</li>
</ol>

<ol>
	<li>Hooton, T. M., Stamm, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+and+treatment+of+uncomplicated+urinary+tract+infection.">Diagnosis and treatment of uncomplicated urinary tract infection.</a> Infect Dis Clin North Am. 11 (3), 551-581 (1997).</li><li>Harper, M., Fowlis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+urinary+tract+infections+in+men.">Management of urinary tract infections in men.</a> Trends in Urology, Gynaecology &amp; Sexual Health. 12 (1), 30-35 (2007).</li><li>Foxman, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+epidemiology+of+urinary+tract+infection.">The epidemiology of urinary tract infection.</a> Nat Rev Urol. 7 (12), 653-660 (2010).</li><li>Lipsky, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Urinary+tract+infections+in+men.+Epidemiology,+pathophysiology,+diagnosis,+and+treatment.">Urinary tract infections in men. Epidemiology, pathophysiology, diagnosis, and treatment.</a> Ann Intern Med. 110 (2), 138-150 (1989).</li><li>Conway, L. J., Carter, E. J., Larson, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Risk+Factors+for+Nosocomial+Bacteremia+Secondary+to+Urinary+Catheter-Associated+Bacteriuria:+A+Systematic+Review.">Risk Factors for Nosocomial Bacteremia Secondary to Urinary Catheter-Associated Bacteriuria: A Systematic Review.</a> Urol Nurs. 35 (4), 191-203 (2015).</li><li>Markle, J. G., Fish, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SeXX+matters+in+immunity.">SeXX matters in immunity.</a> Trends Immunol. 35 (3), 97-104 (2014).</li><li>Yu, C. Y., Whitacre, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex,+MHC+and+complement+C4+in+autoimmune+diseases.">Sex, MHC and complement C4 in autoimmune diseases.</a> Trends Immunol. 25 (12), 694-699 (2004).</li><li>Castelao, J. 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=Gender-+and+smoking-related+bladder+cancer+risk.">Gender- and smoking-related bladder cancer risk.</a> J Natl Cancer Inst. 93 (7), 538-545 (2001).</li><li>Donsky, H., Coyle, S., Scosyrev, E., Messing, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex+differences+in+incidence+and+mortality+of+bladder+and+kidney+cancers:+national+estimates+from+49+countries.">Sex differences in incidence and mortality of bladder and kidney cancers: national estimates from 49 countries.</a> Urol Oncol. 32 (1), 40-e31 (2014).</li><li>Garg, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender+Disparities+in+Hematuria+Evaluation+and+Bladder+Cancer+Diagnosis:+A+Population-Based+Analysis.">Gender Disparities in Hematuria Evaluation and Bladder Cancer Diagnosis: A Population-Based Analysis.</a> J Urol. , (2014).</li><li>Dobruch, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender+and+Bladder+Cancer:+A+Collaborative+Review+of+Etiology,+Biology,+and+Outcomes.">Gender and Bladder Cancer: A Collaborative Review of Etiology, Biology, and Outcomes.</a> Eur Urol. , (2015).</li><li>Hsu, J. 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=Decreased+tumorigenesis+and+mortality+from+bladder+cancer+in+mice+lacking+urothelial+androgen+receptor.">Decreased tumorigenesis and mortality from bladder cancer in mice lacking urothelial androgen receptor.</a> Am J Pathol. 182 (5), 1811-1820 (2013).</li><li>El Behi, 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=An+essential+role+for+decorin+in+bladder+cancer+invasiveness.">An essential role for decorin in bladder cancer invasiveness.</a> EMBO Mol Med. 5 (12), 1835-1851 (2013).</li><li>Foxman, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+urinary+tract+infections:+incidence,+morbidity,+and+economic+costs.">Epidemiology of urinary tract infections: incidence, morbidity, and economic costs.</a> Am J Med. 113, 05S-13S (2002).</li><li>Ingersoll, M. A., Kline, K. A., Nielsen, H. V., Hultgren, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=G-CSF+induction+early+in+uropathogenic+Escherichia+coli+infection+of+the+urinary+tract+modulates+host+immunity.">G-CSF induction early in uropathogenic Escherichia coli infection of the urinary tract modulates host immunity.</a> Cell Microbiol. 10 (12), 2568-2578 (2008).</li><li>Mora-Bau, G., 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=Macrophages+Subvert+Adaptive+Immunity+to+Urinary+Tract+Infection.">Macrophages Subvert Adaptive Immunity to Urinary Tract Infection.</a> PLoS Pathog. 11 (7), e1005044 (2015).</li><li>Oliveira, P. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technical+Report:+Technique+of+Bladder+Catheterization+in+Female+Mice+and+Rats+for+Intravesical+Instillation+in+Models+of+Bladder+Cancer.">Technical Report: Technique of Bladder Catheterization in Female Mice and Rats for Intravesical Instillation in Models of Bladder Cancer.</a> (Conference Title)39th Scand-LAS and ICLAS Joint Meeting. 36, 5-9 (2009).</li><li>Seager, C. 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=Intravesical+delivery+of+rapamycin+suppresses+tumorigenesis+in+a+mouse+model+of+progressive+bladder+cancer.">Intravesical delivery of rapamycin suppresses tumorigenesis in a mouse model of progressive bladder cancer.</a> Cancer Prev Res (Phila). 2 (12), 1008-1014 (2009).</li><li>Hagberg, L., 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=Ascending,+unobstructed+urinary+tract+infection+in+mice+caused+by+pyelonephritogenic+Escherichia+coli+of+human+origin).">Ascending, unobstructed urinary tract infection in mice caused by pyelonephritogenic Escherichia coli of human origin).</a> Infect Immun. 40 (1), 273-283 (1983).</li><li>Olson, P. D., Hruska, K. A., Hunstad, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Androgens+Enhance+Male+Urinary+Tract+Infection+Severity+in+a+New+Model.">Androgens Enhance Male Urinary Tract Infection Severity in a New Model.</a> J Am Soc Nephrol. , (2015).</li><li>Knipper, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interleukin-4+Receptor+alpha+Signaling+in+Myeloid+Cells+Controls+Collagen+Fibril+Assembly+in+Skin+Repair.">Interleukin-4 Receptor alpha Signaling in Myeloid Cells Controls Collagen Fibril Assembly in Skin Repair.</a> Immunity. 43 (4), 803-816 (2015).</li><li>Hung, C. S., Dodson, K. W., Hultgren, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+murine+model+of+urinary+tract+infection.">A murine model of urinary tract infection.</a> Nat Protoc. 4 (8), 1230-1243 (2009).</li><li>Wright, K. J., Seed, P. C., Hultgren, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+intracellular+bacterial+communities+of+uropathogenic+Escherichia+coli+depends+on+type+1+pili.">Development of intracellular bacterial communities of uropathogenic Escherichia coli depends on type 1 pili.</a> Cell Microbiol. 9 (9), 2230-2241 (2007).</li></ol>

作者没有什么可透露的。

经尿道电切的雄性小鼠对膀胱黏膜疾病的影响提供了许多新的研究机会, 但也提出了一些挑战。首先, 一个限制是, 灌输可能最初证明是技术上的困难, 导致过度炎症在导管插入。技术的改进可以通过用有色溶液注入死老鼠来实现, 如埃文斯蓝染料。为了证实注入成功, 膀胱应进行视觉检查和灌输的液体的体积可以用结核菌素注射器的提取评估。缓慢的, 温和的运动的重要性不能被夸大: 不应该有阻力和没有力量用于插入导管, 因为这将导致过多的炎症和组织损伤。当正确地执行时, 导尿管将毫不费力地滑入尿道。如果感觉到阻力或阻塞, 最好是取出导管和尝试插入物。
通过将灌注对细菌负荷的质量与24小时后感染的相关性, 我们证明了一种不完美的技术, 在感染时少量的细菌接种丢失, 仍然会导致强大的定植膀胱.在使用本协议时, 调查人员应致力于实现无泄漏灌注。然而, 该程序的稳健性确保了不完美的灌注仍将提供有用的数据和解释的结果。
我们的技术的巨大优势, 在唯一发表的替代方法的男性膀胱感染是其非侵入性的, 非手术方法。我们使用经尿道电灌注的方法是沿着尿道到膀胱的生理途径, 而不破坏腹部的结构完整性。奥尔森和同事20描述的侵入性技术包括了手术过程中固有的因素, 包括炎症、延迟愈合和瘢痕组织。特别是有关免疫应答感染的研究是机体对外科创伤的反应。这包括形成一个亲炎症的环境和潜在的组织肉芽, 以及抗炎伤口愈合程序的一部分, 愈合过程。这些因素在实验环境中表现出不良影响。
最后, 本研究的目的是开发一种方法, 使男性和女性的免疫应答过程中, 在尿路感染的过程中, 可用于未来的研究, 以解决性别对膀胱黏膜疾病的影响, 如传染病和癌症生物学。作为第二个明显的优势, 这里描述的方法在雌性小鼠中使用了几十年19。因此, 这种技术在雄性小鼠身上进行的实验与现有研究的范围相当广泛。我们的实验表明, 反应的差异确实存在, 这些差异可以提供线索, 以了解基于性别的差异, 对粘膜感染的反应, 以及其他疾病的膀胱。

泌尿道感染是发达国家最常见的感染之一1。在新生儿和老年人中, 女性和男性的感染率相似,2。绝经前的成年女性, 但是, 有一个大大增加的发病率社区获得性尿路感染相比, 男性2,3。鉴于这一疾病主要影响妇女, 基础和临床研究压倒性地集中在女性尿路感染。然而, 在男性泌尿感染是一个重要的和理解的医疗保健挑战4。事实上, 由于男性泌尿系感染与发病率较高有关, 这些传染病通常被定义为复杂的4,5。
随着我们对性别偏见在生理学和病理学中的核心作用的理解, 我们需要新的方法来探索这一以前被忽视的疾病方面。性别差异在免疫和感染中起着至关重要的作用;女性自身免疫性疾病的发病率较高, 而男性更容易受到某些感染, 如结核病、疟疾和 HIV6,7。膀胱癌, 另一个泌尿系病理, 是显着更普遍的男性比妇女, 和几项研究显示了作用的雄激素在发展的恶性肿瘤8,9,10,11 ,12。然而, 值得注意的是, 由于无法反复 catheterize 雄性小鼠13, 对膀胱癌的膀胱灌注治疗的研究完全是在雌性动物身上进行的。
对泌尿学发病机制的研究主要依赖于啮齿动物的感染模型 (如、小鼠和大鼠)。小鼠尿路感染模型可能采用 uropathogens 最初孤立于人类传染病, 如肾炎大肠杆菌(UPEC),克雷伯杆菌,肠球菌,葡萄球菌, 或变形杆菌14。通常情况下, 细菌被引入膀胱通过导尿管的尿道。继感染后, 膀胱和肾脏可以被移除, 以评估特定的感染参数, 如细菌定植、组织损伤或宿主免疫应答15,16。然而, 普遍的, 只有雌性动物被用于泌尿的研究。事实上, 许多研究指出, 由于解剖原因, 男性小鼠的插管是不可能的13,17,18,19。最近, 对男性灌输的外科手术方法已经被描述, 在其中腹部打开, 膀胱是外部流离失所通过施加温和的压力, 在腹部的开放, 细菌被注射到膀胱20。这种方法能使男性感染, 以手术治疗为代价。因此, 这种方法的一个主要的告诫包括炎症的影响, 对感染的结果, 如潜在的诱导抗炎伤口愈合的反应的切口21。由于我们的兴趣包括对疾病的性别偏见的理解, 我们开发了一种在雄性小鼠体内进行细菌膀胱灌注的方法, 它与长期建立的非手术电切法在雌性啮齿动物中的应用更加吻合22.
我们的模型建立在一个既定的方法和提供的能力, 直接比较宿主免疫反应与泌尿系统感染的雌性和雄性动物。这种方法将允许解剖的性别差异的感染, 并可能提供分子和细胞线索的明显差异的敏感性和反应的性别感染。此外, 这一模式的价值超越了泌尿系研究, 允许建立模型, 以调查其他膀胱相关疾病, 如膀胱癌, 前列腺炎, 下或过活动膀胱综合征, 和间质性膀胱炎。

<table><tbody><tr><td>BD Insyte Autoguard Shielded IV Catheters 24 G, 0.7 mm external diameter, 14 mM long</td><td>BD Medical</td><td>381811 or 381411*</td><td>* catalog number is country-dependent</td></tr><tr><td>inoculating loop</td><td>Greiner Bio-One</td><td>731171</td><td></td></tr><tr><td>LB Miller broth</td><td>Difco</td><td>244620</td><td></td></tr><tr><td>LB Miller agar</td><td>Difco</td><td>244520</td><td></td></tr><tr><td>Cuvettes</td><td>Bio-rad</td><td>223-9955</td><td></td></tr><tr><td>syringes</td><td>B Braun</td><td>9166017V</td><td></td></tr><tr><td>Thumb (Adson) forceps</td><td>Fine Science Tools</td><td>11006-12</td><td></td></tr><tr><td>5 ml tubes, polypropylene</td><td>Falcon</td><td>352063</td><td></td></tr><tr><td>Tissue Ruptor disposable probes</td><td>Qiagen</td><td>990890</td><td></td></tr><tr><td>15 ml flip cap tubes</td><td>Thermo Scientific</td><td>362694</td><td></td></tr><tr><td>DNAse</td><td>Invitrogen</td><td>18047019</td><td></td></tr><tr><td>Liberase TM</td><td>Sigma-Aldrich</td><td>5401119001</td><td>should be aliquotted and stored at -20 &amp;deg;C to avoid repeated freeze-thaw</td></tr><tr><td>100 &amp;micro;M MACS SmartStrainers&amp;nbsp;</td><td>Miltenyi</td><td>130-098-463</td><td>compatible with 15 ml tubes, preferred</td></tr><tr><td>100 &amp;micro;m cell strainers&amp;nbsp;</td><td>Falcon</td><td>352360</td><td>compatible with 50 ml tubes, if MACS Smart Strainers are not available</td></tr><tr><td>96 well plate</td><td>Falcon</td><td>353077</td><td></td></tr><tr><td>Fc block</td><td>BD Pharmingen</td><td>553142</td><td>anti-CD16/CD32</td></tr><tr><td>anti-mouse CD45 antibody</td><td>BD Biosciences</td><td>561487</td><td>many different fluorescent conjugation options are available</td></tr><tr><td>5 ml tubes polystyrene with filter cap</td><td>Falcon</td><td>352235</td><td></td></tr><tr><td>insulin syringe</td><td>Terumo</td><td>BS05M2913</td><td></td></tr><tr><td>hand held homogenizer</td><td>Qiagen</td><td>9001272</td><td>TissueRuptor</td></tr><tr><td>flow cytometer</td><td>BD Biosciences</td><td>N/A</td><td>Fortessa SORP&amp;nbsp;</td></tr><tr><td>Flow cytometry software</td><td>BD Biosciences</td><td>N/A</td><td>Diva</td></tr></tbody></table>

urinary tract infection in a small animal model: transurethral catheterization of male and female mice

泌尿道感染在世界范围内非常普遍, 引起严重的发病率和医疗费用。小动物模型, 准确地反映疾病的建立和进展, 允许解剖宿主-病原体相互作用和产生的免疫力感染。在小鼠, 膀胱灌注肾炎的大肠杆菌, 致病剂在85% 以上的社区获得性尿路感染, 重述的许多阶段的感染者观察到的人。然而, 直到最近, 泌尿感染只能以雌性动物为模型。这一限制阻碍了研究泌尿和其他膀胱疾病, 如癌症的性别相关的差异。在这里, 我们描述一种方法来灌输雄性小鼠, 允许直接比较雌性和雄性动物, 并提供一个详细的协议, 以评估膀胱组织的流式细胞仪作为一种手段, 更好地了解宿主对感染的反应。这些方法将有助于确定在泌尿系和其他膀胱相关疾病中所观察到的性别偏见的寄主因素。

在这个协议的发展中, 雌性和雄性 C57Bl/6 小鼠的年龄在6到8周之间被灌输, 并且细菌负担评估在膀胱在时间点范围从1小时到30天。这些研究的结果在别处 (Zychlinsky 沙夫et al., 待定) 进行了详细说明。在这里, 我们提供来自24小时感染的代表性数据。值得注意的是, 24 小时后感染的雄性和雌性小鼠的细菌负担是相等的 (图 1)。在每组24小时感染后观察到的细菌负荷变化不大, 与雌性小鼠的报告相似,15,16。
如果在感染前对尿液进行足够的排空, 雌性小鼠很少会发现接种的严重渗漏。然而, 对雄性小鼠的灌输导致大量的细菌接种从尿道漏出, 特别是在本议定书的发展阶段。为了确定感染时接种的损失是否会影响定植或感染的建立, 我们对每种灌输都采用了评分系统。每一个灌输的评分是 1-5, 1 是最理想的, 细菌定植确定24小时后感染 (图 2, 框)。虽然这个系统是调查者依赖, 因此, 潜在的主观, 本研究的主要作者确定了所有的分数后立即灌输, 在评估细菌的负担在感染膀胱。从不同的灌注分数的动物获得的 CFU 之间没有统计学意义上的差异, 这是由非参数克鲁斯卡尔-瓦利斯试验, 比较每一个列平均秩和每其他列的平均秩的评估 (p = 0.17)用邓恩的测试进行多重比较从这一分析, 可以得出结论, 次优灌注 (评分 > 3), 导致大量泄漏细菌接种在感染期间, 不会显着影响细菌定植在24小时 (图 2,图)。值得注意的是, 根据经验, 雄性小鼠的渗漏频率显著降低。
最后, 开发该模型的目的是直接比较雌性和雄性动物对 UPEC 的免疫应答。通过流式细胞仪测定雌性和雄性小鼠在单纯性和感染人群中的 CD45+免疫细胞的数量, 以检测细胞浸润是否在对泌尿网的反应中发生改变。虽然在天真的动物中存在的免疫细胞的数量在雌性和雄性小鼠之间没有区别 (p = 0.95), 但在感染雌性小鼠的膀胱中有统计学显著的增加 (p = 0.0015) (图 3 a-b)。
<img alt="Figure 1" src="/files/ftp_upload/54432/54432fig1.jpg"> 图 1:UPEC 殖民女性和男性膀胱的效率相同.六到8周的老雌性和雄性 C57Bl/6 小鼠被灌输了 107 CFU UPEC。24小时后感染, 膀胱被菌清除, 以列举细菌的负担。剧情描绘 CFU/膀胱。每个点是一只老鼠, 代表性实验5显示。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54432/54432fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" src="/files/ftp_upload/54432/54432fig2.jpg"> 图 2:灌注质量与24小时感染后的细菌负担无关.六到8周的老雌性和雄性 C57Bl/6 小鼠被灌输了 107 CFU UPEC。在每次灌输后, 一个研究人员会根据特定的标准 (盒装文本) 对灌输的质量评分进行分配。24小时后感染, 膀胱被菌清除, 以列举细菌的负担。剧情描绘了被分配的质量比分与CFU/膀胱。每个点是一只老鼠, 5 汇集实验显示。p = 0.17, 克鲁斯卡尔-沃利斯测试比较每列的平均秩与每隔一列的平均秩, 并对邓恩的测试进行多次比较。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54432/54432fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" src="/files/ftp_upload/54432/54432fig3.jpg"> 图 3:雌性小鼠免疫细胞浸润增加, 以应对尿路感染.六至8周的老雌性和雄性 C57Bl/6 小鼠被灌输或不与 107 CFU UPEC。有代表性的点地块描述总膀胱细胞与 CD45 的+免疫细胞封闭在粉红色的天真或受感染的小鼠。图显示了来自天真或24小时感染小鼠的膀胱中 CD45 的+免疫细胞的绝对数量。每个点是一只老鼠, 2 汇集实验显示。由曼-惠特尼测试确定的 p 值。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54432/54432fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Urinary Tract Infection in a Small Animal Model: Transurethral Catheterization of Male and Female Mice at JoVE.com

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

建立的模型, 经尿道小鼠导尿, 允许研究膀胱疾病, 包括泌尿道感染, 但只能在女性进行。一个新的男性经尿道灌注的模式, 在这里提出, 将使研究在一个地区的特点是强的临床和流行病学差异的性别。

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, ...

urinary-tract-infection-small-animal-model-transurethral

Nanyang Technological University

Research

JoVE Journal

Immunology and Infection

16.4K 次观看.  Institut Pasteur, INSERM U818. 建立的模型, 经尿道小鼠导尿, 允许研究膀胱疾病, 包括泌尿道感染, 但只能在女性进行。一个新的男性经尿道灌注的模式, 在这里提出, 将使研究在一个地区的特点是强的临床和流行病学差异的性别。

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

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

Diagnosis of Ecto- and Endoparasites in Laboratory Rats and Mice

Internal and external parasites remain a significant concern in laboratory rodent facilities, and many research facilities harbor some parasitized animals. Before embarking on an examination of animals for parasites, two things should be considered. One: what use will be made of the information collected, and two: which test is the most appropriate. Knowing that animals are parasitized may be something that the facility accepts, but there is often a need to treat animals and then to determine the efficacy of treatment. Parasites may be detected in animals through various techniques, including samples taken from live or euthanized animals. Historically, the tests with the greatest diagnostic sensitivity required euthanasia of the animal, although PCR has allowed high-sensitivity testing for several types of parasite. This article demonstrates procedures for the detection of endo- and ectoparasites in mice and rats. The same procedures are applicable to other rodents, although the species of parasites found will differ.

This article describes various procedures for screening rats and mice to detect endo- or ectoparasitism. Several diagnostic assays will be demonstrated, both those suitable for use on live animals and those used after euthanasia of the animal. Photographs to aid in identification of rat and mouse parasites will be included.

Ecto和实验室大鼠和小鼠体内寄生虫的诊断

Internal and external parasites remain a significant concern in laboratory rodent facilities, and many research facilities harbor some parasitized ...

A Method for Labeling Vasculature in Embryonic Mice

The establishment of a functional blood vessel network is an essential part of organogenesis, and is required for optimal organ function. For example, in the thymus proper vasculature formation and patterning is essential for thymocyte entry into the organ and mature T-cell exit to the periphery. The spatial arrangement of blood vessels in the thymus is dependent upon signals from the local microenvironment, namely thymic epithelial cells (TEC). Several recent reports suggest that disruption of these signals results in thymus blood vessel defects 1,2. Previous studies have described techniques used to label the neonatal and adult thymus vasculature 1,2. We demonstrate here a technique for labeling blood vessels in the embryonic thymus. This method combines the use of FITC-dextran or Griffonia (Bandeiraea) Simplicifolia Lectin I (GSL 1 - isolectin B4) facial vein injections and CD31 antibody staining to identify thymus vascular structures and PDGFR-&beta; to label thymic perivascular mesenchyme 3-5. The option of using cryosections or vibratome sections is also provided. This protocol can be used to identify thymus vascular defects, which is critical for defining the roles of TEC-derived molecules in thymus blood vessel formation. As the method labels the entire vasculature, it can also be used to analyze the vascular networks in multiple organs and tissues throughout the embryo including skin and heart 6-10.

This article describes a method for labeling embryonic skin and thymus blood vessels.

标签胚胎小鼠血管的方法

The establishment of a functional blood vessel network is an essential part of organogenesis, and is required for optimal organ function. For example, in ...

Establishment and Characterization of UTI and CAUTI in a Mouse Model

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. Females are disproportionately afflicted by UTI: 50% of all women will have a UTI in their lifetime. Additionally, 20-40% of these women who have an initial UTI will suffer a recurrence with some suffering frequent recurrences with serious deterioration in the quality of life, pain and discomfort, disruption of daily activities, increased healthcare costs, and few treatment options other than long-term antibiotic prophylaxis. Uropathogenic Escherichia coli (UPEC) is the primary causative agent of community acquired UTI. Catheter-associated UTI (CAUTI) is the most common hospital acquired infection accounting for a million occurrences in the US annually and dramatic healthcare costs. While UPEC is also the primary cause of CAUTI, other causative agents are of increased significance including Enterococcus faecalis. Here we utilize two well-established mouse models that recapitulate many of the clinical characteristics of these human diseases. For UTI, a C3H/HeN model recapitulates many of the features of UPEC virulence observed in humans including host responses, IBC formation and filamentation. For CAUTI, a model using C57BL/6 mice, which retain catheter bladder implants, has been shown to be susceptible to E. faecalis bladder infection. These representative models are being used to gain striking new insights into the pathogenesis of UTI disease, which is leading to the development of novel therapeutics and management or prevention strategies.

The ability to model urinary tract infections (UTI) is crucial in order to be able to understand bacterial pathogenesis and spawn the development of novel therapeutics. This work&#8217;s goal is to demonstrate mouse models of experimental UTI and catheter associated UTI that recapitulate and predict findings seen in humans.

建立和UTI和CAUTI表征在小鼠模型

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. ...

医学

Transurethral Instillation Procedure in Adult Male Mouse

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into the urinary bladder to either induce animal models of bladder pathologies or evaluate the effectiveness of intravesical treatments. Most rodent models of lower urinary tract (LUT) pathologies are induced in female mice due to ease of intravesical instillation of the substances via the female urethra. However, due to anatomical differences between the female and male LUT, transurethral instillation in a male mouse has been deemed a very challenging procedure, and it has not been previously described. In this manuscript, we provide a detailed description of how to prepare polyethylene (PE) tubing for subsequent insertion into the urethra of a male mouse. In addition, we discuss the ideal types of PE tubing to be used depending on the desired site of inoculation. Furthermore, we describe point by point how to prepare an animal for a successful transurethral instillation to avoid injury to the urethra and ensure the delivery of the solution to the desired location. The procedure is started by retracting the prepuce and the glans to expose the opening of the urethral meatus. Next, the glans are grasped by blunt non-crushing forceps to stabilize the penis and the PE tubing. The PE tubing is first inserted into the urethral meatus parallel to the animal body, then its angle is adjusted by tilting the catheter to maneuver it to follow the natural curvature of the urethra. This technique can be used to induced murine models of bladder pathologies and/or evaluate the effectiveness of intravesical treatments in male mice.

Transurethral instillation is a challenging procedure and has not been well described in the literature. The aim of this manuscript is to describe a technique for transurethral insertion of a catheter for intravesical delivery of liquids with active substances into the urinary bladder and/or prostate in adult male mice.

成年雄性小鼠经尿道灌注术

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into ...

Longitudinal Follow-Up of Urinary Tract Infections and Their Treatment in Mice using Bioluminescence Imaging

Urinary tract infections (UTI) rank among the most common bacterial infections in humans and are routinely treated with empirical antibiotics. However, due to increasing microbial resistance, the efficacy of the most used antibiotics has declined. To find alternative treatment options, there is a great need for a better understanding of the UTI pathogenesis and the mechanisms that determine UTI susceptibility. In order to investigate this in an animal model, a reproducible, non-invasive assay to study the course of UTI is indispensable.
For years, the gold standard for the enumeration of bacterial load has been the determination of Colony Forming Units (CFU) for a particular sample volume. This technique requires post-mortem organ homogenates and serial dilutions, limiting data output and reproducibility. As an alternative, bioluminescence imaging (BLI) is gaining popularity to determine the bacterial load. Labeling pathogens with a lux operon allow for the sensitive detection and quantification in a non-invasive manner, thereby enabling longitudinal follow-up. So far, the adoption of BLI in UTI research remains limited.
This manuscript describes the practical implementation of BLI in a mouse urinary tract infection model. Here, a step-by-step guide for culturing bacteria, intravesical instillation and imaging is provided. The in vivo correlation with CFU is examined and a proof-of-concept is provided by comparing the bacterial load of untreated infected animals with antibiotic-treated animals. Furthermore, the advantages, limitations, and considerations specific to the implementation of BLI in an in vivo UTI model are discussed. The implementation of BLI in the UTI research field will greatly facilitate research on the pathogenesis of UTI and the discovery of new ways to prevent and treat UTI.

This manuscript describes the intravesical administration of uropathogenic bacteria with a lux operon to induce a urinary tract infection in mice and subsequent longitudinal in vivo analysis of the bacterial load using bioluminescence imaging.

使用生物发光成像对小鼠尿路感染及其治疗的纵向随访

Urinary tract infections (UTI) rank among the most common bacterial infections in humans and are routinely treated with empirical antibiotics. However, ...

Evaluating the Anti-Tumor Efficacy of a Bacterial Immunomodulatory Protein in a Murine Model

Source: Codolo, G. et al. <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/52743">Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model.</a>&#160;J. Vis. Exp. (2015)
This video demonstrates a technique to assess the anti-tumor activity of Helicobacter pylori neutrophil-activating protein, or HP-NAP, in a mouse bladder cancer model. HP-NAP interacts with tumor-cell neutrophils to produce pro-inflammatory cytokines, inducing the differentiation of na&#239;ve T cells into type 1 T helper (Th1) and type 1 cytotoxic (Tc1) T cell phenotypes. The Th1 and Tc1 cells secrete interferon-gamma, inhibiting tumor angiogenesis and promoting cytotoxic T cell-mediated tumor destruction, reducing tumor volume.

在小鼠模型中评估细菌免疫调节蛋白的抗肿瘤功效

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

JoVE Encyclopedia of Experiments

Immunology

Immunotherapy

Model Development and Disease Study

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.remotexs.ntu.edu.sg/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

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.remotexs.ntu.edu.sg/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.

雌性小鼠尿路感染模型的建立

Generating Urinary Tract Infection Model: A Technique to Administer Uropathogenic Bacteria in Mouse to Develop Urinary Tract Infection Model

Source: Luyts, N. et al., <a href="https://www-jove-com.remotexs.ntu.edu.sg/v/62614">Longitudinal Follow-Up of Urinary Tract Infections and Their Treatment in Mice using Bioluminescence Imaging.</a> J. Vis. Exp. (2021).
In this video, we describe the technique for the intravesical administration of uropathogenic bacteria to generate a urinary tract infection model in mice. This model can be used to understand the spread of uropathogenic bacterial infection within the urinary system and the mechanisms that determine UTI susceptibility.

生成尿路感染模型:一种在小鼠中施用尿路致病菌以开发尿路感染模型的技术

Source: Luyts, N. et al., Longitudinal Follow-Up of Urinary Tract Infections and Their Treatment in Mice using Bioluminescence Imaging. J. Vis. Exp. ...