The intracellular Ca²⁺ release channel TRPML1 regulates lower urinary tract smooth muscle contractility

TRPML1 (transient receptor potential mucolipin 1) is a Ca²⁺-permeable, nonselective cation channel that is predominantly localized to the membranes of late endosomes and lysosomes (LELs). Intracellular release of Ca²⁺ through TRPML1 is thought to be pivotal for maintenance of intravesicular acidic pH as well as the maturation, fusion, and trafficking of LELs. Interestingly, genetic ablation of TRPML1 in mice (Mcoln1 ) induces a hyperdistended/hypertrophic bladder phenotype. Here, we investigated this phenomenon further by exploring an unconventional role for TRPML1 channels in the regulation of Ca²⁺-signaling activity and contractility in bladder and urethral smooth muscle cells (SMCs). Four-dimensional (4D) lattice light-sheet live-cell imaging showed that the majority of LELs in freshly isolated bladder SMCs were essentially immobile. Superresolution microscopy revealed distinct nanoscale colocalization of LEL-expressing TRPML1 channels with ryanodine type 2 receptors (RyR2) in bladder SMCs. Spontaneous intracellular release of Ca²⁺ from the sarcoplasmic reticulum (SR) through RyR2 generates localized elevations of Ca²⁺ ("Ca²⁺ sparks") that activate plasmalemmal large-conductance Ca²⁺-activated K⁺ (BK) channels, a critical negative feedback mechanism that regulates smooth muscle contractility. This mechanism was impaired in Mcoln1 mice, which showed diminished spontaneous Ca²⁺ sparks and BK channel activity in bladder and urethra SMCs. Additionally, ex vivo contractility experiments showed that loss of Ca²⁺ spark-BK channel signaling in Mcoln1 mice rendered both bladder and urethra smooth muscle hypercontractile. Voiding activity analyses revealed bladder overactivity in Mcoln1 mice. We conclude that TRPML1 is critically important for Ca²⁺ spark signaling, and thus regulation of contractility and function, in lower urinary tract SMCs.