Identification and characterization of TNTs in vitro and in vivo – LiveTuneL

Novel actin-based membrane protrusions named tunneling nanotubes (TNTs) have been found to connect remote cells allowing communication during development, and disease transmission, suggesting that TNTs are ubiquitous in both physiological and pathological contexts. However, their existence in vivo has not been demonstrated and the mechanisms of TNT formation and fusion with a target cell are unknown. This project seeks to understand how protein-protein/protein lipid specific interactions are involved in forming TNTs, and what are the mechanical and dynamical differences distinguishing TNTs from filopodia. Here the involvement of actin, actin-regulating proteins, membrane connectors, and novel candidates identified by a screening approach, will be ascertained in real time in vitro and correlated to mechanical measurements and then tested in vivo in zebrafish embryos. Complementary correlative cryoCLEM will elucidate native TNT structure in different cellular contexts and in the animal.