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* These authors contributed equally
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).
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.
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.....
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
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.......
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, .......
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.
....Name | Company | Catalog Number | Comments |
21mm Clear Tubing | Supera Anesthesia Innov | 301-150 | |
27 gauge needle | BD | Z192376 | |
4 port Manifold | Supera Anesthesia Innov | RES536 | |
DEFINITY | Lantheus Medical Imaging | DE4 | |
F/AIR Canister | Supera Anesthesia Innov | 80120 | |
Graefe forceps (Serrated, Straight) | F.S.T. | 11050-10 | |
Inlet/Outlet Fittings | Supera Anesthesia Innov | VAP203/4 | |
Isoflurane | Midwest Vet Supply | 193.33161.3 | |
Isoflurane Vaporizer | Supera Anesthesia Innov | VAP3000 | |
MV707 probe | Fujifilm VisualSonics Inc | ||
Oxygen Flowmeter | Supera Anesthesia Innov | OXY660 | |
Polyethylene 50 tubing | BD | 427516 | |
Pressure Reg/Gauge | Supera Anesthesia Innov | OXY508 | |
Rebreathing Circuits | Supera Anesthesia Innov | CIR529 | |
Small Mice Nose Cone | Supera Anesthesia Inov | ACC526 | |
Sterile saline | Midwest Vet Supply | 193.74504.3 | NaCl 0.9%, Injectable |
Straight Sharp/Blunt Scissors | Fine Scientific Tools (F.S.T) | 14054-13 | |
Syringe | BD | 309646 | 5mL |
Vevo 770 | Fujifilm VisualSonics Inc | ||
VIALMIX | Lantheus Medical Imaging | VMIX |
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