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A study on etiology of incontinence in double knockout mouse model

  • Priyank Yadav
    Affiliations
    Division of Urology, Department of Surgery, The Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada

    Department of Urology and Renal Transplantation, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
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  • Walid A. Farhat
    Correspondence
    Correspondence to: Walid A Farhat Division of Pediatric Urology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
    Affiliations
    Program in Developmental & Stem Cell Biology, Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada

    Division of Pediatric Urology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA
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  • Adonis Hijaz
    Affiliations
    University Hospitals Cleveland Medical Center, Urology Institute, Cleveland, OH, USA
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  • Jiwon Seo
    Affiliations
    University of Wisconsin School of Medicine and Public Health, Madison, WI, USA
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  • Chi-Chung Hui
    Affiliations
    Program in Developmental & Stem Cell Biology, Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada

    Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada
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  • Karen Tuba-Ang
    Affiliations
    Department of Pathology, Baguio General Hospital and Medical Center, Baguio, Benguet, Philippines
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  • Rong Mo
    Affiliations
    Program in Developmental & Stem Cell Biology, Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada
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  • Michael Chua
    Affiliations
    Division of Urology, Department of Surgery, The Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada

    Institute of Urology, St. Luke's Medical Center, Quezon City, Philippines
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Published:October 05, 2022DOI:https://doi.org/10.1016/j.jpurol.2022.10.002

      Summary

      Introduction and objective

      Stress urinary incontinence is of concern in both pediatric and adult population. Double mutant GLI family zinc finger Gli2+/−; Gli3Δ699/+ murine model of stress incontinence has been recently developed as a reliable model which does not require surgical manipulation to create incontinence and is shown to survive to adulthood. The aim of this study was to establish the etiology of incontinence in the double mutant Gli2+/−; Gli3Δ699/+ mice.

      Study design

      We used 13 cluster of differentiation 1 (CD-1) mice (7–9 weeks) for demonstration of histology of the bladder and urethra. There were 3 Wild Gli2+/- females, 2 Wild Gli2+/- males, 4 Gli2+/-;Gli3Δ699/+ females and 4 Gli2+/-;Gli3Δ699/+ males. The Wild Gli2+/- mice served as the control group and Gli2+/-;Gli3Δ699/+ mice served as the test group. Additionally, eight 16.5 days mice (2 each of Wild Gli2+/- females, Wild Gli2+/- males, double knockout (DKO) Gli2+/-;Gli3Δ699/+ females and Gli2+/-;Gli3Δ699/+ males) were used to assess the histology of the spinal cord. The gross appearance of bladder and urethra was studied using ink injection assays. Immunohistochemistry was done for smooth muscle actin and cytokeratin.

      Results

      Gross and histologic appearance confirmed the previously reported widening of bladder outlet and hypoplasia of smooth muscles in female urethra and also established them in the male urethra of Gli2+/-;Gli3Δ699/+ mice compared to Gli2+/- mice. The double knockout mice were smaller than the Gli2 mice (5.2 vs 6.1 cm, p = 0.002). Immunohistochemistry demonstrated epithelial hyperplasia and smooth muscle hypoplasia. Additionally, there was prostatic hypoplasia in the Gli2+/-;Gli3Δ699/+ male mice. The spinal cord length for body size appeared comparable between the Gli2+/- and Gli2+/-;Gli3Δ699/+ mice but histological evaluation revealed abnormal development of the caudal end of the vertebral body with premature termination of the spinal cord (Figure).

      Discussion

      The histological changes in the bladder neck and urethra were consistent to those previously reported. While previous report described the findings in female mice only, we confirmed that these findings are also present in males as well as prostatic hypoplasia, a possible additional factor leading to stress incontinence. The most important finding in the present study however, was the detection of premature termination of spinal cord in the DKO Gli2+/−; Gli3Δ699/+ mice which has not been reported previously and is likely a major contributor to incontinence in this model.

      Conclusion

      Summary Figure
      Graphical Abstract(a) Gross appearance of the spinal cord in Gli2+/−; Gli3Δ699/+ (left side) and Gli2+/− (right side) mice showing proportional length for body size. (b) Histological evaluation of 16.5 days Gli2+/− (left side) and Gli2+/−; Gli3Δ699/+ (right side) mice showing caudal vertebral bodies and spinal cords. Note the difference at the caudal end (Asterisks denote attachment of tail). Scale bars: 1 mm.

      Keywords

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