Name: transurethral induction of mouse urinary tract infection
Uploaded: 2010-08-05T17:30:00
Description: Watch this Scientific Journal Video about Transurethral Induction of Mouse Urinary Tract Infection at JoVE.com

著者は彼らの専門的な技術支援のためにシェリー謝とジェニファーペインに感謝します。この作業は小児泌尿器科とスタンフォード小児研究基金協会からの助成金を戴いています。

尿道カテーテルのI.準備<ol><li>各マウスの膀胱及びサンプリングする腎臓の場合は、それぞれ1クライオバイアルチューブ5-6ステンレススチール製ビーズ（膀胱、1.6 mmのサイズまたは0.9〜2.0ミリメートルのブレンド）または酸化ジルコニウムビーズ（腎臓、0.5mmのサイズ）を配置。ビーズを含むチューブをオートクレーブ。 </li><li>周囲温度に冷却管の後、各ステンレススチールビーズを含むチ...

<ol>
	<li>Schaeffer, A. J., Schwan, W. R., Hultgren, S. J., Duncan, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+of+type+1+pilus+expression+in+Escherichia+coli+to+ascending+urinary+tract+infections+in+mice.">Relationship of type 1 pilus expression in Escherichia coli to ascending urinary tract infections in mice.</a> Infect Immun. 55, 373-380 (1987).</li><li>Rouschop, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Urothelial+CD44+facilitates+Escherichia+coli+infection+of+the+murine+urinary+tract.">Urothelial CD44 facilitates Escherichia coli infection of the murine urinary tract.</a> J Immunol. 177, 7225-7232 (2006).</li><li>Malaviya, R., Ikeda, T., Abraham, S. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+mast+cells+to+bacterial+clearance+and+their+proliferation+during+experimental+cystitis+induced+by+type+1+fimbriated+E.+coli.">Contribution of mast cells to bacterial clearance and their proliferation during experimental cystitis induced by type 1 fimbriated E. coli.</a> Immunol LettM. 91, 103-111 (2004).</li><li>Lane, M. C., Alteri, C. J., Smith, S. N., Mobley, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+flagella+is+coincident+with+uropathogenic+Escherichia+coli+ascension+to+the+upper+urinary+tract.">Expression of flagella is coincident with uropathogenic Escherichia coli ascension to the upper urinary tract.</a> Proc Natl Acad Sci U S A. 104, 16669-16674 (2007).</li><li>Hvidberg, H. <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+a+long-term+ascending+urinary+tract+infection+mouse+model+for+antibiotic+treatment+studies.">Development of a long-term ascending urinary tract infection mouse model for antibiotic treatment studies.</a> Antimicrob Agents Chemother. 44, 156-163 (2000).</li><li>Hopkins, W. J., Hall, J. A., Conway, B. P., Uehling, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+urinary+tract+infection+by+intraurethral+inoculation+with+Escherichia+coli:+refining+the+murine+model.">Induction of urinary tract infection by intraurethral inoculation with Escherichia coli: refining the murine model.</a> J Infect Dis. 171, 462-465 (1995).</li><li>Hopkins, W. J., Gendron-Fitzpatrick, A., Balish, E., Uehling, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+and+host+responses+to+Escherichia+coli+urinary+tract+infection+in+genetically+distinct+mouse+strains.">Time course and host responses to Escherichia coli urinary tract infection in genetically distinct mouse strains.</a> Infect Immun. 66, 2798-2802 (1998).</li><li>Gunther, N. W. t., Lockatell, V., Johnson, D. E., Mobley, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+dynamics+of+type+1+fimbria+regulation+in+uropathogenic+Escherichia+coli+during+experimental+urinary+tract+infection.">In vivo dynamics of type 1 fimbria regulation in uropathogenic Escherichia coli during experimental urinary tract infection.</a> Infect Immun. 69, 2838-2846 (2001).</li><li>Bishop, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic+AMP-regulated+exocytosis+of+Escherichia+coli+from+infected+bladder+epithelial+cells.">Cyclic AMP-regulated exocytosis of Escherichia coli from infected bladder epithelial cells.</a> Nat Med. 13, 625-630 (2007).</li><li>Thumbikat, P., Waltenbaugh, C., Schaeffer, A. J., Klumpp, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antigen-specific+responses+accelerate+bacterial+clearance+in+the+bladder.">Antigen-specific responses accelerate bacterial clearance in the bladder.</a> J Immunol. 176, 3080-3086 (2006).</li><li>Hagberg, L. <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, 273-283 (1983).</li><li>Hopkins, W. J., Hall, J. A., Conway, B. P., Uehling, D. 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細菌の経尿道的接種によるマウス尿路感染症の誘導は、老舗が技術的な要求手順11です。ハングとHultgrenは最近、マウス尿路感染症13の経尿道的技術の優れたレビューを発表した。本稿では、さらにこの実験的なアプローチの細かい点を説明しよう。 初心者は、膀胱を公開する正中開腹術を行うインドインク（PBS中の1％）の希釈溶液を注入し、膀胱の青色...

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		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Cryovial tubes, 2.0 ml polypropylene (no caps)</td>
<td>&nbsp;</td>
<td>E&amp;K Scientific</td>
<td>640201</td>
<td>&nbsp;</td>
<tr>
<td>Caps for cryovial tubes, with 'O' ring</td>
<td>&nbsp;</td>
<td>E&amp;K Scientific</td>
<td>440011-N</td>
<td>&nbsp;</td>
<tr>
<td>Bullet Blender</td>
<td>&nbsp;</td>
<td> Next Advance</td>
<td>BBX24</td>
<td>&nbsp;</td>
<tr>
<td>Zirconium Oxide Beads (0.5mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>ZROB05</td>
<td>&nbsp;</td>
<tr>
<td>Stainless Steel Beads (1.6mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>SSB16</td>
<td>&nbsp;</td>
<tr>
<td>Stainless Steel Bead Blend (0.9-2.0mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>SSB14B</td>
<td>&nbsp;</td>
<tr>
<td>CBA/J Mice, 8-12 week old female</td>
<td>&nbsp;</td>
<td>Jackson Laboratories</td>
<td>000656</td>
<td>&nbsp;</td>
<tr>
<td>Phosphate Buffered Saline (NaCl 137mM, KCl 2.7mM, Na2HPO4 4.3mM, KH2PO4 1.4mM)</td>
<td>&nbsp;</td>
<td>EMD</td>
<td>EM-SX0420-13 (NaCl) EM-PX1405-1 (KCl) EM1.06585.0 (Na2HPO4) EM-5108-1 (KH2PO4)</td>
<td>&nbsp;</td>
<tr>
<td>LB agar</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>LB broth</td>
<td>&nbsp;</td>
<td>EMD</td>
<td>1.10285.0500</td>
<td>&nbsp;</td>
<tr>
<td>Isoflurane (Aerrane)</td>
<td>&nbsp;</td>
<td>Baxter</td>
<td>NDC 10019-773-60</td>
<td>&nbsp;</td>
<tr>
<td>Inoculating loops</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>22-363-601</td>
<td>&nbsp;</td>
<tr>
<td>Eosin-methylene blue (EMB) agar powder</td>
<td>&nbsp;</td>
<td>Himedia Laboratories</td>
<td>MM022-500G</td>
<td>&nbsp;</td>
<tr>
<td>MacConkey agar powder</td>
<td>&nbsp;</td>
<td>Difco</td>
<td>212123</td>
<td>&nbsp;</td>
<tr>
<td>LB agar powder</td>
<td>&nbsp;</td>
<td>Difco</td>
<td>244520</td>
<td>&nbsp;</td>
<tr>
<td>Intramedic non-radiopaque polyethylene tubing, PE10 [inner dimension 0.28 mm (0.011 inch); outer dimension 0.61 mm (0.024 inch)]</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>427401</td>
<td>&nbsp;</td>
<tr>
<td>30 gauge needles, 1/2 inch</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>305106</td>
<td>&nbsp;</td>
<tr>
<td>1 ml tuberculin Slip-Tip syringe</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>309602</td>
<td>&nbsp;</td>
<tr>
<td>3 ml Luer-Lock syringe</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>309585</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

transurethral induction of mouse urinary tract infection

興味の尿路病原性菌株は、寒天上で増殖されています。一般的に、尿路疾患<em&gt; E.大腸菌</em&gt;（UPEC）と他の株は37℃ルリア - ベルターニ（LB）寒天培地上で一晩成長させることができる·周囲空気中のC。 UPECの菌株は、LB寒天培地上で黄色がかった白色半透明のコロニーとして成長する。適切なコロニー形態の確認後、シングルコロニーを培養液で培養されるに応じて選択されます。 LBブロスは、ほとんどの尿路病原性菌株に使用することができます。二つのシリアル、一晩LBブロス培養物はI型線毛、尿路病原性の細菌のための明確に定義された毒性因子の発現を増強するために用いることができる。ブロス培養物（PBS）リン酸緩衝生理食塩水で所望の濃度に希釈されています。 8〜12週齢の雌マウスは、イソフルラン麻酔下に置き、経尿道的にポリエチレンチューブで覆われた30ゲージのシリンジを使用して細菌を接種されています。経験的にそれぞれの細菌/マウス系統の組み合わせのために決定しなければならない典型的な接種は、10です。<sup&gt; 6</sup&gt; 10〜<sup&gt; 8</sup&gt; CFU PBSの10〜50マイクロリットルのマウスあたり。希望の感染期間（数週間に一日）、尿サンプルと膀胱と両方の腎臓が収穫された後。各臓器は、細かく刻むPBSに入れ、そしてブルーブレットホモジナイザーで均質化する。尿および組織ホモジネートが直列にPBSで希釈し、適切な寒天上で培養されています。翌日は、コロニー形成単位がカウントされます。

Watch this Scientific Journal Video about Transurethral Induction of Mouse Urinary Tract Infection at JoVE.com

Transurethral Induction of Mouse Urinary Tract Infection

このビデオでは、経尿道的にマウスの尿路感染症を誘発し、結果として感染症の程度を定量化する方法を紹介します。

マウス尿路感染症の経尿道的誘導

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

Research

JoVE Journal

Immunology and Infection

Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis

実験的自己免疫性下垂体炎の誘導のためのマウス下垂体免疫原の調製

Autoimmune hypophysitis is a chronic inflammation of the pituitary gland caused or accompanied by autoimmunity1.  It has traditionally been considered a ...

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

SJLマウスにおける実験的自己免疫性下垂体炎の誘導

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse ...

RNA Isolation of Pseudomonas aeruginosa Colonizing the Murine Gastrointestinal Tract

RNA Isolation of Pseudomonas aeruginosa Colonizing the Murine Gastrointestinal Tract

のRNAの単離緑膿菌マウス消化管に定着

Pseudomonas aeruginosa (PA) infections result in significant morbidity and mortality in hosts with compromised immune systems, such as patients with ...

Isolation of Lymphocytes from Mouse Genital Tract Mucosa

マウス生殖管粘膜からのリンパ球の単離

Mucosal surfaces, including in the gastrointestinal, urogenital, and respiratory tracts, provide portals of entry for pathogens, such as viruses and ...

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

小動物モデルにおける尿路感染症: 雌雄マウスの尿道カテーテル

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

Transurethral Instillation Procedure in Adult Male Mouse

大人の雄のマウスに経尿道的注入手順

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

Mouse Models Of Helicobacter Infection And Gastric Pathologies

Mouse Models Of Helicobacter Infection And Gastric Pathologies

ヘリコバクター ・ ピロリ感染と胃の病理のマウス ・ モデル

Helicobacter pylori is a gastric pathogen that is present in half of the global population and is a significant cause of morbidity and mortality in ...

Isolation of Dendritic Cells from the Human Female Reproductive Tract for Phenotypical and Functional Studies

人間の女性の生殖器官から樹状細胞の表現型および機能研究の分離

The characterization of the human dendritic cells (DCs) resident in mucosal tissues is challenging due to the difficulty in obtaining samples, and the low ...

Isolation of Single Intracellular Bacterial Communities Generated from a Murine Model of Urinary Tract Infection for Downstream Single-cell Analysis

下流の単一細胞分析のため尿路感染症のマウスモデルから生成された単一細胞内細菌群集の分離

In this article, we outline a procedure used to isolate individual intracellular bacterial communities from a mouse that has been experimentally infected ...

Induction of Mouse Lung Injury by Endotracheal Injection of Bleomycin

ブレオマイシンの気管内注射によるマウス肺損傷の誘導

Pulmonary fibrosis is a hallmark of several human lung diseases with a different etiology. Since current therapies are rather limited, mouse models ...