MSP-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...

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Main Authors: Jennifer H Richens, Mariia Dmitrieva, Helen L Zenner, Nadine Muschalik, Richard Butler, Jade Glashauser, Carolina Camelo, Stefan Luschnig, Sean Munro, Jens Rittscher, Daniel St Johnston
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-04-01
Series:PLoS Biology
Online Access:https://doi.org/10.1371/journal.pbio.3003099
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author Jennifer H Richens
Mariia Dmitrieva
Helen L Zenner
Nadine Muschalik
Richard Butler
Jade Glashauser
Carolina Camelo
Stefan Luschnig
Sean Munro
Jens Rittscher
Daniel St Johnston
author_facet Jennifer H Richens
Mariia Dmitrieva
Helen L Zenner
Nadine Muschalik
Richard Butler
Jade Glashauser
Carolina Camelo
Stefan Luschnig
Sean Munro
Jens Rittscher
Daniel St Johnston
author_sort Jennifer H Richens
collection DOAJ
description 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-Golgi vesicles, revealing that Cad99c vesicles predominantly move apically with a mean speed of 1.1µm/sec. This is reduced to 0.85 µm/sec by a dominant slow dynein mutant, demonstrating that dynein transports Cad99C vesicles to the apical cortex. Furthermore, both the dynein mutant and microtubule depolymerization cause lateral Cad99C secretion. Thus, microtubule organization plays a central role in targeting apical secretion, suggesting that Drosophila does not have distinct apical versus basolateral vesicle fusion machinery. Nidogen vesicles undergo planar-polarized transport to the leading edge of follicle cells as they migrate over the ECM, whereas most Collagen is secreted at trailing edges. The follicle cells therefore bias secretion of different ECM components to opposite sides of the cell, revealing that the secretory pathway is more spatially organized than previously thought.
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spelling doaj-art-4abf9e35048f412ea2d84b3eb5a4e4212025-08-23T05:30:55ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852025-04-01234e300309910.1371/journal.pbio.3003099MSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.Jennifer H RichensMariia DmitrievaHelen L ZennerNadine MuschalikRichard ButlerJade GlashauserCarolina CameloStefan LuschnigSean MunroJens RittscherDaniel St JohnstonUnderstanding 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-Golgi vesicles, revealing that Cad99c vesicles predominantly move apically with a mean speed of 1.1µm/sec. This is reduced to 0.85 µm/sec by a dominant slow dynein mutant, demonstrating that dynein transports Cad99C vesicles to the apical cortex. Furthermore, both the dynein mutant and microtubule depolymerization cause lateral Cad99C secretion. Thus, microtubule organization plays a central role in targeting apical secretion, suggesting that Drosophila does not have distinct apical versus basolateral vesicle fusion machinery. Nidogen vesicles undergo planar-polarized transport to the leading edge of follicle cells as they migrate over the ECM, whereas most Collagen is secreted at trailing edges. The follicle cells therefore bias secretion of different ECM components to opposite sides of the cell, revealing that the secretory pathway is more spatially organized than previously thought.https://doi.org/10.1371/journal.pbio.3003099
spellingShingle Jennifer H Richens
Mariia Dmitrieva
Helen L Zenner
Nadine Muschalik
Richard Butler
Jade Glashauser
Carolina Camelo
Stefan Luschnig
Sean Munro
Jens Rittscher
Daniel St Johnston
MSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.
PLoS Biology
title MSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.
title_full MSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.
title_fullStr MSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.
title_full_unstemmed MSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.
title_short MSP-tracker: A versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in Drosophila epithelial cells.
title_sort msp tracker a versatile vesicle tracking software tool used to reveal the spatial control of polarized secretion in drosophila epithelial cells
url https://doi.org/10.1371/journal.pbio.3003099
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