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

Los autores desean agradecer a Shelly Xie y Jennifer Payne por su asistencia técnica de expertos. Este trabajo fue posible gracias a donaciones de la Sociedad de Urología Pediátrica y el Fondo de Stanford Pediatric Research.

I. Preparación de sondas uretrales <ol><li> Para cada uno de vejiga y los riñones del ratón para el muestreo, lugar 5-6 gotas de acero inoxidable (vejiga, el tamaño de 1,6 mm o de 0,9 a 2,0 mm mezcla) o granos de óxido de circonio (de los riñones, el tamaño de 0,5 mm) en un tubo de criovial, respectivamente. Autoclave de los tubos de cuentas que contienen. </li><li> Después de los tubos de enfriar a temperatura ambiente, añadir 200 microlitros de PBS estéril a cada cordón de acero inoxidable que contiene el...

<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. 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La inducción de la infección del tracto urinario del ratón mediante la inoculación de bacterias transuretral es un procedimiento de larga data, pero técnicamente exigente 11. Colgado y Hultgren ha publicado recientemente una excelente revisión de las técnicas transuretral de infección del tracto urinario del ratón 13. En este trabajo, se intenta ilustrar los puntos más importantes de este enfoque experimental. Los principiantes pueden practicar su técnica med...

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		<div>
		<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

Cepas bacterianas uropatógena de interés se cultivan en agar. En general, los uropatógenos<em&gt; E. coli</em&gt; (UPEC) y otras cepas pueden ser cultivadas durante la noche en Luria-Bertani (LB) de agar a 37 ° C en el aire ambiente. Cepas UPEC crecer como colonias de color blanco amarillento translúcido en LB agar. Tras la confirmación de la morfología de las colonias apropiadas, las colonias solo entonces se escogió para ser cultivadas en caldo. Caldo de LB se pueden utilizar para la mayoría de cepas bacterianas uropatógenos. Dos culturas de serie, la noche de caldo de LB se pueden emplear para aumentar la expresión de pili de tipo I, un factor de virulencia bien definidas para las bacterias uropatógenos. Caldos de cultivo se diluye a la concentración deseada en tampón fosfato salino (PBS). De ocho a 12 semanas de edad ratones hembra se colocan bajo anestesia con isoflurano y transuretral inoculadas con las bacterias mediante tubos de polietileno cubiertas de 30 jeringas de calibre. Inóculos típico, el cual debe ser determinada empíricamente para cada combinación de cepas bacterianas / ratón, son 10<sup&gt; 6</sup&gt; A 10<sup&gt; 8</sup&gt; UFC por ratón de 10 a 50 microlitros de PBS. Tras el periodo de infección deseado (un día a varias semanas), las muestras de orina y la vejiga y los riñones tanto se cosechan. Cada órgano es picado, colocado en PBS, y se homogeneizaron en un homogeneizador Bala Azul. Homogeneizados de orina y el tejido se diluyen en serie en PBS y se cultivaron en agar apropiado. Al día siguiente, las unidades formadoras de colonias se cuentan.

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

Transurethral Induction of Mouse Urinary Tract Infection

Este video se muestran los métodos para inducir transuretral infecciones del tracto urinario ratón y cuantificar la extensión de las infecciones resultantes.

Inducción transuretral de infección del tracto urinario ratón

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

Preparación de inmunógeno ratón hipófisis para la inducción de la hipofisitis autoinmune experimental

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

Investigación

Inmunología e Infección

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

La inducción de la hipofisitis autoinmune experimental en ratones SJL

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

El aislamiento de ARN Pseudomonas aeruginosa Que colonizan el tracto gastrointestinal murino

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

Aislamiento de linfocitos de ratón del tracto genital Mucosa

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

Infección del tracto urinario en un modelo Animal pequeño: cateterización transuretral de ratones machos y hembras

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

Procedimiento transuretral instilación en ratón macho adulto

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

Modelos de ratón de la infección por Helicobacter y patologías gástricas

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

Aislamiento de las células dendríticas del tracto reproductivo femenino humano estudios fenotípica y funcional

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

Aislamiento de las comunidades bacterianas intracelulares solo generados a partir de un modelo murino de infección del tracto urinario para posteriores análisis unicelular

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

Inducción de lesión pulmonar de ratón por inyección endotraqueal de bleomicina

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