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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

We describe the use of high frequency ultrasound with contrast imaging as a method to measure bladder volume, bladder wall thickness, urine velocity, void volume, void duration, and urethral diameter. This strategy can be used to assess voiding dysfunction and treatment efficacy in various mouse models of lower urinary tract dysfunction (LUTD).

Abstract

The incidence of clinical benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTS) is increasing due to the aging population, resulting in a significant economic and quality of life burden. Transgenic and other mouse models have been developed to recreate various aspects of this multifactorial disease; however, methods to accurately quantitate urinary dysfunction and the effectiveness of new therapeutic options are lacking. Here, we describe a method that can be used to measure bladder volume and detrusor wall thickness, urinary velocity, void volume and void duration, and urethral diameter. This would allow for the evaluation of disease progression and treatment efficacy over time. Mice were anesthetized with isoflurane, and the bladder was visualized by ultrasound. For non-contrast imaging, a 3D image was taken of the bladder to calculate volume and evaluate shape; the bladder wall thickness was measured from this image. For contrast-enhanced imaging, a catheter was placed through the dome of the bladder using a 27-gauge needle connected to a syringe by PE50 tubing. A bolus of 0.5 mL of contrast was infused into the bladder until a urination event occurred. Urethral diameter was determined at the point of the Doppler velocity sample window during the first voiding event. Velocity was measured for each subsequent event yielding a flow rate. In conclusion, high frequency ultrasound proved to be an effective method for assessing bladder and urethral measurements during urinary function in mice. This technique may be useful in the assessment of novel therapies for BPH/LUTS in an experimental setting.

Introduction

Benign prostatic hyperplasia (BPH) is a disease that develops in men as they age and affects nearly 90% of men over 80 years of age1,2. Although the development of BPH is generally associated with aging, other factors including obesity and metabolic syndrome can lead to BPH in relatively younger men3,4. Many men with BPH develop lower urinary tract symptoms (LUTS) that significantly decrease their quality of life, and some experience complications that may include bleeding, infection, bladder outlet obstruction (BOO), bladder stones, and renal failure.....

Protocol

Procedures involving animal subjects have been approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Wisconsin – Madison.

1. Animal preparation

  1. Place a 24-month-old, C57Bl6/J male mouse in a pre-charged chamber with 3-5% isoflurane until the righting reflex is lost and the breathing rate slows.
  2. If necessary, use clippers to shave the abdominal hair from the animal for surgery and/or imaging. Remove all remaining hair using a depila.......

Representative Results

Ultrasound can be used with or without contrast enhancement depending on the experimental design and endpoint measurement. Mice are anesthetized with isoflurane and shaved and all traces of hair removed with a depilatory cream. Anesthetized animals are placed on a heated platform with the ultrasound probe positioned along the long axis of the bladder (Figure 1).

Figure 2 shows representative ultrasound images of a mouse bladder acquir.......

Discussion

Current techniques for evaluating the lower urinary tract of rodents are limited by their ability to directly correlate changes in voiding physiology with changes in prostatic histology consequent to disease progression. Void spot assays and uroflowmetry can be used to assess spontaneous urination events in rodents, and these techniques can be used to evaluate changes over a period of time15,16,17. However, for both techniques, .......

Acknowledgements

We would like to thank Emily Ricke, Kristen Uchtmann, and the Ricke lab for their assistance with animal husbandry and feedback on this manuscript. We would like to thank the NIDDK and NIEHS for their financial support for these studies: U54 DK104310 (WAR, JAM, PCM, CMV, DEB), R01 ES001332 (WAR, CMV), K12 DK100022 (TTL, AR-A, DH). The content is the sole responsibility of the authors and does not represent the official views of the NIH.

....

Materials

NameCompanyCatalog NumberComments
21mm Clear TubingSupera Anesthesia Innov301-150
27 gauge needleBDZ192376
4 port ManifoldSupera Anesthesia InnovRES536
DEFINITYLantheus Medical ImagingDE4
F/AIR CanisterSupera Anesthesia Innov80120
Graefe forceps (Serrated, Straight)F.S.T.11050-10
Inlet/Outlet FittingsSupera Anesthesia InnovVAP203/4
IsofluraneMidwest Vet Supply193.33161.3
Isoflurane VaporizerSupera Anesthesia InnovVAP3000
MV707 probeFujifilm VisualSonics Inc
Oxygen FlowmeterSupera Anesthesia InnovOXY660
Polyethylene 50 tubingBD427516
Pressure Reg/GaugeSupera Anesthesia InnovOXY508
Rebreathing CircuitsSupera Anesthesia InnovCIR529
Small Mice Nose ConeSupera Anesthesia InovACC526
Sterile salineMidwest Vet Supply193.74504.3NaCl 0.9%, Injectable
Straight Sharp/Blunt ScissorsFine Scientific Tools (F.S.T)14054-13
SyringeBD3096465mL
Vevo 770Fujifilm VisualSonics Inc
VIALMIXLantheus Medical ImagingVMIX

References

  1. Kirby, R. S. The natural history of benign prostatic hyperplasia: what have we learned in the last decade. Urology. 5, 3-6 (2000).
  2. Berry, S. J., Coffey, D. S., Walsh, P. C., Ewing, L. L. The develo....

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