Giant organelle vesicles to uncover intracellular membrane mechanics and plasticity

Alexandre Santinho, Maxime Carpentier, Julio Lopes Sampaio, Mohyeddine Omrane and Abdou Rachid Thiam ()
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Alexandre Santinho: Université PSL, CNRS, Sorbonne Université, Université Paris Cité
Maxime Carpentier: Université PSL, CNRS, Sorbonne Université, Université Paris Cité
Julio Lopes Sampaio: PSL Research University, Plateforme de Métabolomique et Lipidomique, 26 rue d’Ulm
Mohyeddine Omrane: Université PSL, CNRS, Sorbonne Université, Université Paris Cité
Abdou Rachid Thiam: Université PSL, CNRS, Sorbonne Université, Université Paris Cité

Nature Communications, 2024, vol. 15, issue 1, 1-12

Abstract: Abstract Tools for accessing and studying organelles remain underdeveloped. Here, we present a method by which giant organelle vesicles (GOVs) are generated by submitting cells to a hypotonic medium followed by plasma membrane breakage. By this means, GOVs ranging from 3 to over 10 µm become available for micromanipulation. GOVs are made from organelles such as the endoplasmic reticulum, endosomes, lysosomes and mitochondria, or in contact with one another such as giant mitochondria-associated ER membrane vesicles. We measure the mechanical properties of each organelle-derived GOV and find that they have distinct properties. In GOVs procured from Cos7 cells, for example, bending rigidities tend to increase from the endoplasmic reticulum to the plasma membrane. We also found that the mechanical properties of giant endoplasmic reticulum vesicles (GERVs) vary depending on their interactions with other organelles or the metabolic state of the cell. Lastly, we demonstrate GERVs’ biochemical activity through their capacity to synthesize triglycerides and assemble lipid droplets. These findings underscore the potential of GOVs as valuable tools for studying the biophysics and biology of organelles.

Date: 2024
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DOI: 10.1038/s41467-024-48086-7

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