Regulation of Mechanosensor PIEZO Channel Trafficking in C. elegans Germline

Abstract

PIEZO proteins are essential mechanotransductive ion channels, yet mechanisms governing their subcellular localization and membrane trafficking remain elusive. Here, we leverage the C. elegans germline as an in vivo imaging platform to decipher the molecular networks directing PIEZO channel dynamics. High-resolution confocal imaging of endogenously tagged PEZO-1, the C. elegans ortholog, reveals that PEZO-1 compartmentalizes with caveolae protein CAV-1 and the recycling endosome regulator RAB-11. Depletion of rab-11 or the t-SNARE syx-4 severely impairs the translocation of PEZO-1-positive vesicles to the plasma membrane, establishing a reliance on conserved, RAB-11-dependent machinery. Furthermore, we demonstrated that PEZO-1 structural integration is essential for vesicle formation and transport; truncating either the N-terminal transmembrane domains or the C-terminal ion pore induces aberrant vesicle morphology and arrests trafficking. Crucially, PEZO-1 vesicle formation and transport are non-autonomously modulated by reproductive signals requiring male sperm or major sperm protein (MSP) signaling. Finally, introducing conserved disease-associated pathogenic PIEZO mutations markedly suppresses its cytosolic and plasma membrane expression. Collectively, our findings define a fundamental cytological framework regulating PIEZO channel dynamics, shedding light on the molecular etiology of PIEZO-associated channelopathies.