biostudies-literatureRichens JHCancer Research UKDeutsche ForschungsgemeinschaftWellcome TrustBiotechnology and Biological Sciences Research Councile3003099https://www.ebi.ac.uk/biostudies/studies/S-EPMC12021295biostudies-literatureUnknown23(4)Understanding how specific secretory cargoes are targeted to distinct domains of the plasma membrane in epithelial cells requires analyzing the trafficking of post-Golgi vesicles to their sites of secretion. We used the RUSH (retention using selective hooks) system to synchronously release an apical cargo, Cadherin 99C (Cad99C), and a basolateral cargo, the ECM protein Nidogen, from the endoplasmic reticulum and followed their movements to the plasma membrane. We also developed an interactive vesicle tracking framework, MSP-tracker and viewer, that exploits developments in computer vision and deep learning to determine vesicle trajectories in a noisy environment without the need for extensive training data. MSP-tracker outperformed other tracking software in detecting and tracking post-GolPLoS biologyMSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.PMC12021295A24823203144203285095927SFB 1009092096207496SFB 1348080007A14492BB/P026486/1Richens JHButler RCamelo CMuschalik NGlashauser JZenner HLRittscher JDmitrieva MLuschnig SMunro SSt Johnston DfalseMSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.Understanding how specific secretory cargoes are targeted to distinct domains of the plasma membrane in epithelial cells requires analyzing the trafficking of post-Golgi vesicles to their sites of secretion. We used the RUSH (retention using selective hooks) system to synchronously release an apical cargo, Cadherin 99C (Cad99C), and a basolateral cargo, the ECM protein Nidogen, from the endoplasmic reticulum and followed their movements to the plasma membrane. We also developed an interactive vesicle tracking framework, MSP-tracker and viewer, that exploits developments in computer vision and deep learning to determine vesicle trajectories in a noisy environment without the need for extensive training data. MSP-tracker outperformed other tracking software in detecting and tracking post-Gol2025-01-01T00:00:00Z2025 Apr2026-05-29T19:04:00.338Z2025-08-12T03:04:48.289ZS-EPMC120212954020890110.1371/journal.pbio.3003099