TRPML1 channels initiate Ca ²⁺ sparks in vascular smooth muscle cells

Abstract

TRPML1 (transient receptor potential mucolipin 1) is a Ca²⁺-permeable, nonselective cation channel localized to the membranes of endosomes and lysosomes and is not present or functional on the plasma membrane. Ca²⁺ released from endosomes and lysosomes into the cytosol through TRPML1 channels is vital for trafficking, acidification, and other basic functions of these organelles. Here, we investigated the function of TRPML1 channels in fully differentiated contractile vascular smooth muscle cells (SMCs). In live-cell confocal imaging studies, we found that most endosomes and lysosomes in freshly isolated SMCs from cerebral arteries were essentially immobile. Using nanoscale super-resolution microscopy, we found that TRPML1 channels present in late endosomes and lysosomes formed stable complexes with type 2 ryanodine receptors (RyR2) on the sarcoplasmic reticulum (SR). Spontaneous Ca²⁺ signals resulting from the release of SR Ca²⁺ through RyR2s ("Ca²⁺ sparks") and corresponding Ca²⁺-activated K⁺ channel activity are critically important for balancing vasoconstriction. We found that these signals were essentially absent in SMCs from TRPML1-knockout (<i>Mcoln1^-/-</i> ) mice. Using ex vivo pressure myography, we found that loss of this critical signaling cascade exaggerated the vasoconstrictor responses of cerebral and mesenteric resistance arteries. In vivo radiotelemetry studies showed that <i>Mcoln1^-/-</i> mice were spontaneously hypertensive. We conclude that TRPML1 is crucial for the initiation of Ca²⁺ sparks in SMCs and the regulation of vascular contractility and blood pressure.