Name: measurement of tactile allodynia in a murine model of bacterial prostatitis
Uploaded: 2013-01-16T13:00:00
Description: Watch this Scientific Journal Video about Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis at JoVE.com

This research was supported by grant 1K01DK079019A2 (P.T.) from the NIH/NIDDK.

1. Bacteria Preparation for Mouse Inoculation
The following must be done under aseptic conditions.
<ol>
	<li>Take one 17 x 100 mm polypropylene tubes with caps and add 3 ml of fresh Luria broth (LB) media. Using autoclaved tips, take some frozen bacteria glycerol stock of strain CP12 and transfer a small quantity of the culture into the tube containing the LB media. Replace the tube cap so that oxygen is still able to enter the tube - the culture needs to grow under aerobic con...

<ol>
	<li>Yamamoto, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+epidemiology+of+uropathogenic+Escherichia+coli.">Molecular epidemiology of uropathogenic Escherichia coli.</a> Journal of infection and chemotherapy official journal of the Japan Society of Chemotherapy. 13, 68-73 (2007).</li><li>Rudick, C. N., 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=Uropathogenic+Escherichia+coli+induces+chronic+pelvic+pain.">Uropathogenic Escherichia coli induces chronic pelvic pain.</a> Infect. Immun. 79, 628-635 (2011).</li><li>Pontari, M. A., Ruggieri, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+in+prostatitis/chronic+pelvic+pain+syndrome.">Mechanisms in prostatitis/chronic pelvic pain syndrome.</a> The Journal of urology. 172, 839-845 (2004).</li><li>Schaeffer, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+practice.+Chronic+prostatitis+and+the+chronic+pelvic+pain+syndrome.">Clinical practice. Chronic prostatitis and the chronic pelvic pain syndrome.</a> The New England journal of medicine. 355, 1690-1698 (2006).</li><li>Rudick, C. N., Schaeffer, A. J., Thumbikat, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+autoimmune+prostatitis+induces+chronic+pelvic+pain.+American+journal+of+physiology.">Experimental autoimmune prostatitis induces chronic pelvic pain. American journal of physiology.</a> Regulatory, integrative and comparative physiology. 294, R1268-R1275 (2008).</li><li>Elkahwaji, J. E., Ott, C. J., Janda, L. M., Hopkins, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+for+acute+bacterial+prostatitis+in+genetically+distinct+inbred+strains.">Mouse model for acute bacterial prostatitis in genetically distinct inbred strains.</a> Urology. 66, 883-887 (2005).</li><li>Hultgren, S. J., Porter, T. N., Schaeffer, A. 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=Role+of+type+1+pili+and+effects+of+phase+variation+on+lower+urinary+tract+infections+produced+by+Escherichia+coli.">Role of type 1 pili and effects of phase variation on lower urinary tract infections produced by Escherichia coli.</a> Infect. Immun. 50, 370-377 (1985).</li><li>Jarrell, J., Giamberardino, M. A., Robert, M., Nasr-Esfahani, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bedside+testing+for+chronic+pelvic+pain:+discriminating+visceral+from+somatic+pain.">Bedside testing for chronic pelvic pain: discriminating visceral from somatic pain.</a> Pain research and treatment. 2011, 692102 (2011).</li><li>Jarrell, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Demonstration+of+Cutaneous+Allodynia+in+Association+with+Chronic+Pelvic+Pain.">Demonstration of Cutaneous Allodynia in Association with Chronic Pelvic Pain.</a> J. Vis. Exp. (28), e1232 (2009).</li><li>Giamberardino, M. 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=Viscero-visceral+hyperalgesia:+characterization+in+different+clinical+models.">Viscero-visceral hyperalgesia: characterization in different clinical models.</a> Pain. 151, 307-322 (2010).</li><li>Laird, J. M., Martinez-Caro, L., Garcia-Nicas, E., Cervero, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+model+of+visceral+pain+and+referred+hyperalgesia+in+the+mouse.">A new model of visceral pain and referred hyperalgesia in the mouse.</a> Pain. 92, 335-342 (2001).</li><li>Laird, J. M., Souslova, V., Wood, J. N., Cervero, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deficits+in+visceral+pain+and+referred+hyperalgesia+in+Nav1.8+(SNS/PN3)-null+mice.">Deficits in visceral pain and referred hyperalgesia in Nav1.8 (SNS/PN3)-null mice.</a> J. Neurosci. 22, 8352-8356 (2002).</li><li>Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M., Yaksh, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assessment+of+tactile+allodynia+in+the+rat+paw.">Quantitative assessment of tactile allodynia in the rat paw.</a> Journal of neuroscience. 53, 55-63 (1994).</li><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>Boehm, B. J., Colopy, S. A., Jerde, T. J., Loftus, C. J., Bushman, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+bacterial+inflammation+of+the+mouse+prostate.">Acute bacterial inflammation of the mouse prostate.</a> The Prostate. 72, 307-317 (2012).</li><li>Rivero, V. E., Cailleau, C., Depiante-Depaoli, M., Riera, C. M., Carnaud, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-obese+diabetic+(NOD)+mice+are+genetically+susceptible+to+experimental+autoimmune+prostatitis+(EAP).">Non-obese diabetic (NOD) mice are genetically susceptible to experimental autoimmune prostatitis (EAP).</a> Journal of autoimmunity. 11, 603-610 (1998).</li><li>Pakarainen, T., Zhang, F. P., Makela, S., Poutanen, M., Huhtaniemi, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testosterone+replacement+therapy+induces+spermatogenesis+and+partially+restores+fertility+in+luteinizing+hormone+receptor+knockout+mice.">Testosterone replacement therapy induces spermatogenesis and partially restores fertility in luteinizing hormone receptor knockout mice.</a> Endocrinology. 146, 596-606 (2005).</li><li>Taguchi, O., Kojima, A., Nishizuka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+autoimmune+prostatitis+after+neonatal+thymectomy+in+the+mouse.">Experimental autoimmune prostatitis after neonatal thymectomy in the mouse.</a> Clinical and experimental immunology. 60, 123-129 (1985).</li></ol>

Infection of the mouse prostate with UPEC allows for the in vivo modeling of events that may be involved in the pathogenesis of bacterial prostatitis, CP/CPPS or as a predisposing event in chronic inflammation. The methods described for bacterial preparation and instillation draw upon a large body of literature on UPEC models in female urinary tract infection7,14. The model has wide applicability for studying pathogenesis, testing potential vaccine candidates and mechanisms of immune modulation. The a...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 </tr>
 <tr>
 <td>Culture tubes</td>
 <td>BD Falcon</td>
 <td>352059</td>
 </tr>
 <tr>
 <td>LB Broth Miller</td>
 <td>EMD</td>
 <td>EM1.10285.0500 </td>
 </tr>
 <tr>
 <td>Nalgene Centrifuge Tubes</td>
 <td>Thermo</td>
 <td>3118-0050</td>
 </tr>
 <tr>
 <td>Isoflurane</td>
 <td>Butler Schein</td>
 <td>NDC 11695-6776-1</td>
 </tr>
 <tr>
 <td>Catheter needle 30G</td>
 <td>Hamilton</td>
 <td>91030</td>
 </tr>
 <tr>
 <td>PE10 tubing</td>
 <td>BD intramedic</td>
 <td>427400</td>
 </tr>
 <tr>
 <td>Von Frey Filaments</td>
 <td>Stoelting</td>
 <td>58025-31</td>
 </tr>
 </tbody>
</table>

measurement of tactile allodynia in a murine model of bacterial prostatitis

Uropathogenic Escherichia coli (UPEC) are pathogens that play an important role in urinary tract infections and bacterial prostatitis1. We have recently shown that UPEC have an important role in the initiation of chronic pelvic pain2, a feature of Chronic prostatitis/Chronic pelvic pain syndrome (CP/CPPS)3,4. Infection of the prostate by clinically relevant UPEC can initiate and establish chronic pain through mechanisms that may involve tissue damage and the initiation of mechanisms of autoimmunity5.
A challenge to understanding the pathogenesis of UPEC in the prostate is the relative inaccessibility of the prostate gland to manipulation. We utilized a previously described intraurethral infection method6 to deliver a clinical strain of UPEC into male mice thereby establishing an ascending infection of the prostate. Here, we describe our protocols for standardizing the bacterial inoculum7 as well as the procedure for catheterizing anesthetized male mice for instillation of bacteria.
CP/CPPS is primarily characterized by the presence of tactile allodynia4. Behavior testing was based on the concept of cutaneous hyperalgesia resulting from referred visceral pain8-10. An irritable focus in visceral tissues reduces cutaneous pain thresholds allowing for an exaggerated response to normally non-painful stimuli (allodynia). Application of normal force to the skin result in abnormal responses that tend to increase with the intensity of the underlying visceral pain. We describe methodology in NOD/ShiLtJ mice that utilize von Frey fibers to quantify tactile allodynia over time in response to a single infection with UPEC bacteria.

We examined NOD/ShiLtj and C57BL/6J male mice for the presence of chronic pain upon bacterial infection. Male mice (5 to 7 weeks old) were instilled with saline or bacteria into the urethra (Figure 1). Mechanical stimulation of the pelvic area with von Frey filaments of saline-instilled C57BL/6J mice and UPEC-infected C57BL/6J mice resulted in a response frequency that did not change during the 28-day course of the experiment (Figure 1). In contrast, UPEC-infected NOD mice exhibited resp...

Watch this Scientific Journal Video about Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis at JoVE.com

Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis

Infection of the prostate may be a contributing factor in mediating pelvic pain in chronic prostatitis. We describe the procedure for preparation of standardized bacterial inoculum, instillation of bacteria into the urethra of male mice and methodology for measuring tactile allodynia in mice over time.

Uropathogenic Escherichia coli (UPEC) are pathogens that play an important role in urinary tract infections and bacterial prostatitis1. We have recently ...

Research

JoVE Journal

Immunology and Infection

Introducing Shear Stress in the Study of Bacterial Adhesion

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

Induction of Graft-versus-host Disease and In Vivo T Cell Monitoring Using an MHC-matched Murine Model

Induction of Graft-versus-host Disease and In Vivo T Cell Monitoring Using an MHC-matched Murine Model

Induktion af graft-versus-host sygdom og<em&gt; In Vivo</em&gt; T Cell Monitoring Brug en MHC-matchet murin model

Graft-versus-host disease (GVHD) is the limiting barrier to the broad use of bone marrow transplant as a curative therapy for a variety of hematological ...

Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants

Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants

Generation og Multi-fænotypisk High-indhold Screening af<em&gt; Coxiella burnetii</em&gt; Transposon Mutants

Invasion and colonization of host cells by bacterial pathogens depend on the activity of a large number of prokaryotic proteins, defined as virulence ...

A Murine Model of Group B Streptococcus Vaginal Colonization

A Murine Model of Group B Streptococcus Vaginal Colonization

En murin model af gruppe B<em&gt; Streptococcus</em&gt; Vaginal Colonization

Streptococcus agalactiae (group B Streptococcus, GBS), is a Gram-positive, asymptomatic colonizer of the human gastrointestinal tract and vaginal tract of ...

In Vivo Imaging of Reactive Oxygen Species in a Murine Wound Model

In Vivo Imaging of Reactive Oxygen Species in a Murine Wound Model

In Vivo Billeddannelse af reaktive ilt arter i en Murine sår Model

The generation of reactive oxygen species (ROS) is a hallmark of inflammatory processes, but in excess, oxidative stress is widely implicated in various ...

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

Isolering af enkelt intracellulære bakterielle samfund genereret fra en Murine Model af urinvejsinfektion for Downstream encellede analyse

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

A High-throughput Shigella-specific Bactericidal Assay

A High-throughput Shigella-specific Bactericidal Assay

En høj overførselshastighed Shigella-specifikke bakteriedræbende Assay

Serum bactericidal assays (SBAs) measure the functional activity of antibodies and have been used for many decades. SBAs directly measure antibody killing ...

A Precise Pathogen Delivery and Recovery System for Murine Models of Secondary Bacterial Pneumonia

En præcis patogen levering og nyttiggørelse system for murine modeller af sekundær bakteriel lungebetændelse

Secondary bacterial pneumonias following influenza infections consistently rank within the top ten leading causes of death in the United States. To date, ...

Measurement of Pulse Propagation Velocity, Distensibility and Strain in an Abdominal Aortic Aneurysm Mouse Model

Måling af pulse formeringshastighed, distensibility og stamme i en abdominal aorta Aneurisme Mus Model

An abdominal aortic aneurysm (AAA) is defined as a localized dilation of the abdominal aorta that exceeds the maximal intraluminal diameter (MILD) by 1.5 ...

Visualization of Bacterial Resistance using Fluorescent Antibiotic Probes

Visualisering af bakteriel resistens ved hjælp af fluorescerende antibiotika sonder

Fluorescent antibiotics are multipurpose research tools that are readily used for the study of antimicrobial resistance, due to their significant ...