Non-invasive voiding assessment in conscious mice
OBJECTIVE: To review available options of assessing murine bladder function and to evaluate a non-invasive technique suitable for long-term recording.
METHODS: We reviewed previously described methods to record rodent bladder function. We used modified metabolic cages to capture novel recording tracings of mouse micturition. We evaluated our method in a pilot study with female mice undergoing partial bladder outlet obstruction or sham operation, respectively; half of the partial obstruction and sham group received treatment with an S6K-inhibitor, targeting the mTOR pathway, which is known to be implicated in bladder response to obstruction.
RESULTS: Our non-invasive method using continuous urine weight recording reliably detected changes in murine bladder function resulting from partial bladder outlet obstruction or treatment with S6K-inhibitor. We found obstruction as well as treatment with S6K-inhibitor to correlate with a hyperactive voiding pattern.
CONCLUSIONS: While invasive methods to assess murine bladder function largely disturb bladder histology and intrinsically render post-cystometry gene expression analysis of questionable value, continuous urine weight recording is a reliable, inexpensive, and critically non-invasive method to assess murine bladder function, suitable for a long-term application.
1. Andersson K-E, Soler R, Füllhase C. Rodent models for urodynamic investigation. Neurourol Urodynam. Wiley Subscription Services, Inc., A Wiley Company; 2011 Jun 9;30(5):636–46.
2. Bjorling DE, Wang Z-Y, Bushman W. Models of inflammation of the lower urinary tract. Neurourol Urodynam. Wiley Subscription Services, Inc., A Wiley Company; 2011 Jun 1;30(5):673–82.
3. Fry CH, Daneshgari F, Thor K, Drake M, Eccles R, Kanai AJ, et al. Animal models and their use in understanding lower urinary tract dysfunction. Neurourol Urodynam. 2010 Apr;29(4):603–8.
4. Wood R, Eichel L, Messing EM, Schwarz E. Automated noninvasive measurement of cyclophosphamide-induced changes in murine voiding frequency and volume. JURO. 2001 Feb;165(2):653–9.
5. Chen Q, TAKAHASHI S, ZHONG S, HOSODA C. Function of the lower urinary tract in mice lacking α 1d-adrenoceptor. Journal of Experimental Medicine. 2005.
6. Hicks AN, Campeau L, Burmeister D, Bishop CE, Andersson K-E. Lack of nicotinamide mononucleotide adenylyltransferase 2 ( Nmnat2): Consequences for mouse bladder development and function. Neurourol Urodynam. 2013 Jan 31;32(8):1130–6.
7. Campeau L, Füllhase C, Sawada N, Gratzke C, Hedlund P, Howlett AC, et al. Characterization of bladder function in a cannabinoid receptor type 2 knockout mouse in vivo and in vitro. Neurourol Urodynam. 2013 Jul 3;33(5):566–70.
8. Stewart FA, Michael BD, Denekamp J. Late radiation damage in the mouse bladder as measured by increased urination frequency. Radiat Res. 1978 Sep;75(3):649–59.
9. Negoro H, Kanematsu A, Doi M, Suadicani SO, Matsuo M, Imamura M, et al. Involvement of urinary bladder Connexin43 and the circadian clock in coordination of diurnal micturition rhythm. Nat Comms. 2012 May 1;3:809–23.
10. Desjardins C, Maruniak JA, Bronson FH. Social Rank in House Mice: Differentiation Revealed by Ultraviolet Visualization of Urinary Marking Patterns. Science. 1973;182(4115):939–41.
11. Takezawa K, Kondo M, Kiuchi H, Soda T, Takao T, Miyagawa Y, et al. Combination of bladder ultrasonography and novel cystometry method in mice reveals rapid decrease in bladder capacity and compliance in LPS-induced cystitis. Am J Physiol Renal Physiol. American Physiological Society; 2014 Jul 15;307(2):F234–41.
12. Morikawa K, Kakiuchi M, Fukuoka M, KATO H, ITO Y, GOMI Y. Effects of Various Drugs on Bladder Function in Conscious Restrained-Denervated Rats Placed in a Restraining Cage and Produced by Transection of the Hypogastric Nerve. JpnJPharmacol. The Japanese Pharmacological Society; 1990;52(3):405–11.
13. Aitken KJ, Tolg C, Panchal T, Leslie B, Yu J, Elkelini M, et al. Mammalian Target of Rapamycin (mTOR) Induces Proliferation and De-Differentiation Responses to Three Coordinate Pathophysiologic Stimuli (Mechanical Strain, Hypoxia, and Extracellular Matrix Remodeling) in Rat Bladder Smooth Muscle. The American Journal of Pathology. American Society for Investigative Pathology; 2010 Nov 24;176(1):304–19.
14. Schröder A, Kirwan TP, Jiang JX, Aitken KJ, Bägli DJ. Rapamycin Attenuates Bladder Hypertrophy During Long-Term Outlet Obstruction In Vivo: Tissue, Matrix and Mechanistic Insights. JURO. Elsevier Inc; 2013 Jun 1;189(6):2377–84.
15. Dörr W. Cystometry in Mice—Influence of Bladder Filling Rate and Circadian Variations in Bladder Compliance. The Journal of Urology. 1992 Jul;148(1):183–7.
16. Herrera GM, Meredith AL. Diurnal Variation in Urodynamics of Rat. Yamazaki S, editor. PLoS ONE. Public Library of Science; 2010 Aug 19;5(8):e12298–7.
17. Sidler M, Aitken KJ, Jiang J-X, Bägli DJ. Nerve-sparing Mid-urethral Obstruction (NeMO) in Female Small Rodents. JoVE. 2017 Apr 25;(122):e55288–8.
18. Di R, Wu X, Chang Z, Zhao X, Feng Q, Lu S, et al. S6K inhibition renders cardiac protection against myocardial infarction through PDK1 phosphorylation of Akt. Biochem J. Portland Press Limited; 2012 Jan 1;441(1):199–207.
19. Uvelius B, Gabella G. Intramural neurones appear in the urinary bladder wall following excision of the pelvic ganglion in the rat. NeuroReport. 1995;6(16):2213–6.
20. Cornelissen LL, Misajet B, Brooks DP, Hicks A. Influence of genetic background and gender on bladder function in the mouse. Autonomic Neuroscience. 2008 Jun;140(1-2):53–8.
21. Negoro H, Kanematsu A, Matsuo M, Okamura H, Tabata Y, Ogawa O. Development of Diurnal Micturition Pattern in Mice After Weaning. JURO. Elsevier Inc; 2013 Feb 1;189(2):740–6.
22. Moss MC, Rezan T, Karaman UR, Gomelsky A. Treatment of Concomitant OAB and BPH. Curr Urol Rep. Current Urology Reports; 2017 Jan 11;:1–7.
23. Sidler M, Aitken K, Jiang J, Bijos D, Belik J, Bägli DJ. Finding NeMO-Nerve-sparing Mid-urethral Obstruction: A Pathophysiologically Accurate Model of Rodent Partial Bladder Outlet Obstruction. Urology. 2017 Mar 30;105:208.e1–208.e9.
24. Sugino Y, Kanematsu A, Hayashi Y, Haga H, Yoshimura N, Yoshimura K, et al. Voided stain on paper method for analysis of mouse urination. Neurourol Urodynam. Wiley Subscription Services, Inc., A Wiley Company; 2008 Aug;27(6):548–52.
25. Bjorling DE, Wang Z, Vezina CM, Ricke WA, Keil KP, Yu W, et al. Evaluation of voiding assays in mice: impact of genetic strains and sex. Am J Physiol Renal Physiol. 2015 Jun 15;308(12):F1369–78.
26. Mucignat-Caretta C, Bondı̀ M, Caretta A. Endocrine status affects bladder size and postvoid residual urinary volume in mice. Hormones and Behavior. 2004 Jun;46(1):11–8.
27. Abrams P, Cardozo L, Fall M, Griffiths D, Rosier P, Ulmsten U, et al. The standardisation of terminology in lower urinary tract function: report from the standardisation sub-committee of the International Continence Society. Urology. 2003 Jan;61(1):37–49.
28. Winters JC, Dmochowski RR, Goldman HB, Herndon CDA, Kobashi KC, Kraus SR, et al. Urodynamic Studies in Adults: AUA/SUFU Guideline. JURO. Elsevier Inc; 2012 Dec 1;188(S):2464–72.
29. Guven A, Kalorin C, Onal B, Whitbeck C, Chichester P, Kogan BA, et al. Novel biomarkers of bladder decompensation after partial bladder obstruction. Neurourol Urodynam. 2007;26(7):1036–42.
30. Biers SM, REYNARD JM, DOORE T, Brading AF. The functional effects of a c-kit tyrosine inhibitor on guinea-pig and human detrusor. BJU International. 2006 Mar;97(3):612–6.
31. Pandita RK, Fujiwara M, Alm P, Andersson KE. Cystometric evaluation of bladder function in non-anesthetized mice with and without bladder outlet obstruction. JURO. 2000 Oct;164(4):1385–9.
32. Smith PP, Kuchel GA. Continuous uroflow cystometry in the urethane-anesthetized mouse. Neurourol Urodynam. Wiley Subscription Services, Inc., A Wiley Company; 2010 Feb 1;29(7):1344–9.
33. Lundbeck F, Djurhuus JC, Vaeth M. Bladder filling in mice: an experimental in vivo model to evaluate the reservoir function of the urinary bladder in a long term study. The Journal of Urology. 1989.
34. Yaksh TL, Durant PA, Brent CR. Micturition in rats: a chronic model for study of bladder function and effect of anesthetics. Am J Physiol. 1986 Dec;251(6 Pt 2):R1177–85.
35. Baskin LS, Sutherland RS, Thomson AA, Nguyen HT, Morgan DM, Hayward SW, et al. Growth factors in bladder wound healing. JURO. 1997 Jun;157(6):2388–95.
36. Yu W, Ackert-Bicknell C, Larigakis JD, MacIver B, Steers WD, Churchill GA, et al. Spontaneous voiding by mice reveals strain-specific lower urinary tract function to be a quantitative genetic trait. Am J Physiol Renal Physiol. American Physiological Society; 2014 Jun 1;306(11):F1296–307.
37. Keil KP, Abler LL, Altmann HM, Bushman W, Marker PC, Li L, et al. Influence of animal husbandry practices on void spot assay outcomes in C57BL/6J male mice. Neurourol Urodynam. 2014 Nov 12;35(2):192–8.
38. Chen H, Zhang L, Hill WG, Yu W. Evaluating the Voiding Spot Assay in Mice: A Simple Method With Complex Environmental Interactions. Am J Physiol Renal Physiol. American Physiological Society; 2017 Aug 23;:ajprenal.00318.2017–28.
39. Wada N, Shimizu T, Takai S, Shimizu N, Kanai AJ, Tyagi P, et al. Post-injury bladder management strategy influences lower urinary tract dysfunction in the mouse model of spinal cord injury. Neurourol Urodynam. 2016 Oct 24;:1–5.
40. Ikeda Y, Zabbarova I, Schaefer CM, Bushnell DS, de Groat WC, Kanai AJ, et al. Fgfr2 is integral for bladder mesenchyme patterning and function. Am J Physiol Renal Physiol. American Physiological Society; 2017 Jan 4;:ajprenal.00463.2016–44.
41. Takezawa K, Kondo M, Kiuchi H, Ueda N, Soda T, Fukuhara S, et al. Authentic role of ATP signaling in micturition reflex. Sci Rep. Nature Publishing Group; 2016 Jan 6;:1–9.
42. Sano T, Kobayashi T, Negoro H, Sengiku A, Hiratsuka T, Kamioka Y, et al. Intravital imaging of mouse urothelium reveals activation of extracellular signal-regulated kinase by stretch-induced intravesical release of ATP. Physiol Rep. Physiological Reports; 2016 Nov;4(21):e13033–18.