Question

An ex vivo bladder model with detrusor smooth muscle removed to analyse biologically active mediators released from the suburothelium

Answer 1

In order to conduct research on the availability of urothelium-derived mediators at the luminal and anti-luminal features of the urothelium during filling, a decentralized (ex vivo) detrusor smooth muscle (DSM)-denuded mouse bladder preparation was developed.

• C57BL6/J mice had their bladders removed, and the DSM was taken out using a fine-scissor dissection technique without disturbing the mucosa.
• Control, protamine sulfate- and lipopolysaccharide-treated denuded bladders were assessed for their fluorescent dye permeability, prepared wall morphology and cell composition, pressure-volume correlations during filling, and more.
• The DSM and serosa were absent from the preparatory wall, but the urothelium and suburothelium (SubU)/lamina propria (LP) were in good condition.
• By evaluating the release, metabolism, and transport of purine mediators at SubU/LP and in the bladder lumen during filling, the model's applicability for physiological studies was confirmed.
• We discovered asymmetrical purine availability (such as ATP, ADP, AMP, and adenosine) during filling in the lumen and at SubU/LP, indicating distinct purine release, degradation, and bilateral transurothelial transport pathways.
• Some findings were confirmed in a cynomolgus monkey with a DSM-denuded bladder (Macaca fascicularis).
• By allowing direct access to the area around SubU/LP during real bladder filling, the novel model was superior to existing models used to study the properties of the urothelium (such as cultured urothelial cells, bladder mucosa sheets mounted in Ussing chambers, or isolated bladder strips in organ baths).
• The model is especially well-suited for comprehending regional mechanisms of urothelium-DSM connection as well as for a more comprehensive knowledge of the function of urothelium in controlling continence and urination.

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