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

Glashauser, Jade; Camelo, Carolina; Hollmann, Manuel; Backer, Wilko; Jacobs, Thea; Sanchez, Jone Isasti; Schleutker, Raphael; Förster, Dominique; Berns, Nicola; Riechmann, Veit; Luschnig, Stefan

doi:10.1016/j.devcel.2023.03.006

Cited by 5 publications

(3 citation statements)

References 67 publications

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“…Cad99c accumulates at a similar rate at the TGN and at the plasma membrane with a time lag of 6-8 minutes, indicating that this is how long it takes to traffic protein from the TGN to the cell surface. Cad99c reaches the plasma membrane at a similar rate to that of Serpentine in the fly tracheal epithelium but is significantly faster than cargoes that have been tracked in tissue culture cells using RUSH or VSVG-ts [6,14,44]. This may be because the follicle cells and trachea are intact secretory epithelia rather than transformed tissue culture cells and are therefore more efficient at polarised exocytosis.…”

Section: Resultsmentioning

confidence: 99%

Tracking exocytic vesicle movements reveals the spatial control of secretion in epithelial cells

Richens,

Dmitrieva,

Zenner

et al. 2024

Preprint

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Understanding how specific secretory cargoes are targeted to distinct domains of the plasma membrane in epithelial cells requires analysing the trafficking of post-Golgi vesicles to their sites of secretion. Here, we introduce an interactive vesicle tracking framework, the multiscale particle tracker and viewer, that exploits developments in computer vision and deep learning to determine vesicle trajectories in noisy cellular environments without the need for extensive training data. We analysed the movements of secreted cargoes in the Drosophila follicular epithelium ex vivo with high temporal and spatial resolution and found that MSP-tracker outperformed other tracking software. Using the RUSH (retention using selective hooks) system to synchronously release cargoes from the endoplasmic reticulum, we followed the movements the microvillar protein, Cadherin 99C, to the plasma membrane. A dominant slow dynein mutant reduces the speed of apical Cad99C movements, demonstrating that dynein transports Cad99C vesicles to the apical cortex. Furthermore, both the dynein mutant and microtubule depolymerisation cause lateral Cad99C secretion. Thus, microtubule organisation plays a central role in targeting secretion, suggesting that Drosophila does not have distinct apical versus basolateral vesicle fusion machinery. Imaging the extracellular matrix protein (ECM) proteins, Nidogen and Collagen IV revealed that Nidogen vesicles undergo planar-polarised 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 secrete different ECM components at opposite sides of the cell, revealing that the secretory pathway is more spatially organised than previously thought.

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Section: Resultsmentioning

confidence: 99%

Tracking exocytic vesicle movements reveals the spatial control of secretion in epithelial cells

Richens,

Dmitrieva,

Zenner

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…More recently, the implementation of methods to manipulate protein secretion in time and space has further increased the tool box to study morphogens in vivo. [86,87] Protein binder tools can also be used to re-construct a morphogen system in order to address the minimum requirement of morphogen gradient formation. [88,89] However, it is important to note that protein binders may cause leakage or artifacts that need to be taken into consideration.…”

Section: Biological and Experimental Relevance For Other Morphogen Sy...mentioning

confidence: 99%

Is Drosophila Dpp/BMP morphogen spreading required for wing patterning and growth?

Matsuda

Affolter

2023

BioEssays

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Secreted signaling molecules act as morphogens to control patterning and growth in many developing tissues. Since locally produced morphogens spread to form a concentration gradient in the surrounding tissue, spreading is generally thought to be the key step in the non‐autonomous actions. Here, we review recent advances in tool development to investigate morphogen function using the role of decapentaplegic (Dpp)/bone morphogenetic protein (BMP)‐type ligand in the Drosophila wing disc as an example. By applying protein binder tools to distinguish between the roles of Dpp spreading and local Dpp signaling, we found that Dpp signaling in the source cells is important for wing patterning and growth but Dpp spreading from this source cells is not as strictly required as previously thought. Given recent studies showing unexpected requirements of long‐range action of different morphogens, manipulating endogenous morphogen gradients by synthetic protein binder tools could shed more light on how morphogens act in developing tissues.

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“…An additional advantage of researching secretion in Drosophila is the availability of sophisticated tools for transgenic tagging and tissue-specific functional interrogation. Many recent studies have taken advantage of these to dissect secretory traffic in an animals ( Fujii et al, 2020 ; Glashauser et al, 2023 ; Johnson et al, 2020 ; Ma et al, 2020 ; Park et al, 2022 ; Song et al, 2022 ; van Leeuwen et al, 2018 ; Zajac and Horne-Badovinac, 2022 ; Zhou et al, 2023 ). In Drosophila , the early secretory pathway is organized into secretory units, tens to hundreds per cell, in which ERES lie in close proximity to Golgi ministacks ( Kondylis and Rabouille, 2009 ).…”

Section: Introductionmentioning

confidence: 99%

p24–Tango1 interactions ensure ER–Golgi interface stability and efficient transport

Yang,

Feng,

Pastor-Pareja

2024

Journal of Cell Biology

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The eukaryotic p24 family, consisting of α-, β-, γ- and δ-p24 subfamilies, has long been known to be involved in regulating secretion. Despite increasing interest in these proteins, fundamental questions remain about their role. Here, we systematically investigated Drosophila p24 proteins. We discovered that members of all four p24 subfamilies are required for general secretion and that their localizations between ER exit site (ERES) and Golgi are interdependent in an α→βδ→γ sequence. We also found that localization of p24 proteins and ERES determinant Tango1 requires interaction through their respective GOLD and SH3 lumenal domains, with Tango1 loss sending p24 proteins to the plasma membrane and vice versa. Finally, we show that p24 loss expands the COPII zone at ERES and increases the number of ER–Golgi vesicles, supporting a restrictive role of p24 proteins on vesicle budding for efficient transport. Our results reveal Tango1–p24 interplay as central to the generation of a stable ER–Golgi interface.

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

Cited by 5 publications

References 67 publications

Tracking exocytic vesicle movements reveals the spatial control of secretion in epithelial cells

Tracking exocytic vesicle movements reveals the spatial control of secretion in epithelial cells

Is Drosophila Dpp/BMP morphogen spreading required for wing patterning and growth?

p24–Tango1 interactions ensure ER–Golgi interface stability and efficient transport

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