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Acute manipulation and real-time visualization of membrane trafficking and exocytosis in Drosophila

Intracellular trafficking of secretory proteins plays key roles in animal development and physiology, but so far, tools for investigating the dynamics of membrane trafficking have been limited to cultured cells. Here, we present a system that enables acute manipulation and real-time visualization of...

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Main Authors: Glashauser, Jade (Author) , Camelo, Carolina (Author) , Hollmann, Manuel (Author) , Backer, Wilko (Author) , Jacobs, Thea (Author) , Sanchez, Jone Isasti (Author) , Schleutker, Raphael (Author) , Förster, Dominique (Author) , Berns, Nicola (Author) , Riechmann, Veit (Author) , Luschnig, Stefan (Author)
Format: Article (Journal)
Language:English
Published: 24 April 2023
In: Developmental cell
Year: 2023, Volume: 58, Issue: 8, Pages: 709-723
ISSN:1878-1551
DOI:10.1016/j.devcel.2023.03.006
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.devcel.2023.03.006
Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S1534580723001028
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Author Notes:Jade Glashauser, Carolina Camelo, Manuel Hollmann, Wilko Backer, Thea Jacobs, Jone Isasti Sanchez, Raphael Schleutker, Dominique Förster, Nicola Berns, Veit Riechmann, Stefan Luschnig
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Summary:Intracellular trafficking of secretory proteins plays key roles in animal development and physiology, but so far, tools for investigating the dynamics of membrane trafficking have been limited to cultured cells. Here, we present a system that enables acute manipulation and real-time visualization of membrane trafficking through the reversible retention of proteins in the endoplasmic reticulum (ER) in living multicellular organisms. By adapting the “retention using selective hooks” (RUSH) approach to Drosophila, we show that trafficking of GPI-linked, secreted, and transmembrane proteins can be controlled with high temporal precision in intact animals and cultured organs. We demonstrate the potential of this approach by analyzing the kinetics of ER exit and apical secretion and the spatiotemporal dynamics of tricellular junction assembly in epithelia of living embryos. Furthermore, we show that controllable ER retention enables tissue-specific depletion of secretory protein function. The system is broadly applicable to visualizing and manipulating membrane trafficking in diverse cell types in vivo.
Item Description:Online veröffentlicht: 5. April 2023, Artikelversion: 24. April 2023
Gesehen am 20.03.2024
Physical Description:Online Resource
ISSN:1878-1551
DOI:10.1016/j.devcel.2023.03.006

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