Multiple Lipid Scramblases Regulate the Quantity of Extracellular Vesicle Production From the Plasma Membrane.
- 1Department of Biochemistry & Biophysics Texas A&M University College Station Texas USA.
- 2Department of Biological Sciences University of Denver Denver Colorado USA.
- 3Department of Physics & Astronomy University of Denver Denver Colorado USA.
Abstract
Extracellular vesicle (EV) biogenesis is linked to the disruption of lipid bilayer asymmetry during diverse processes from coagulation to programmed cell death. However, the specific contributions of lipid scramblase proteins to the regulation of EV production and the potential redundancy of diverse families of scramblases are not well understood. We developed a semi-automated image analysis pipeline that accurately quantifies fluorescently labelled EVs on the cell surface of live Caenorhabditis elegans embryos. Using this method in a sensitized background with enhanced EV production, we performed an RNA interference (RNAi) screen targeting a subset of plasma membrane and intracellular scramblases. We found that knockdown of most scramblase genes we tested resulted in a minor but significant reduction in the number of EVs. We confirmed these significant decreases in EV abundance with anoh-2 and atg-9 scramblase mutants. However, loss-of-function egli-1 or scrm-3 scramblase mutants displayed no significant change in EV production, unlike egli-1 or scrm-3 knockdown. We also observed no effect on EV production in ced-7 ABC transporter mutants. The discrepancy between the phenotypes of acute scramblase knockdown and chronic loss suggested the presence of compensatory mechanisms that maintain EV homeostasis in response to the permanent absence of a scramblase. RNA sequencing revealed changes in expression levels for conserved sets of genes in egli-1 and scrm-3 mutants. Together, our data suggest that multiple scramblases involved in diverse pathways regulate EV budding, potentially responding to distinct physiological or metabolic cues.
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BibTeX
@article{guerra2026multiple,
title = {{Multiple Lipid Scramblases Regulate the Quantity of Extracellular Vesicle Production From the Plasma Membrane.}},
author = {Bianca L Guerra and Tanner Chase White and Alissa Rumin and Lauren Pitts and Alexander Nguyen and Vanshika Chauhan and Dinah Loerke and Ann M Wehman},
journal = {Journal of extracellular biology},
year = {2026},
month = nov,
doi = {10.1002/jex2.70197},
url = {https://europepmc.org/article/MED/42996306},
note = {CC BY-NC},
}
RIS
TY - JOUR
TI - Multiple Lipid Scramblases Regulate the Quantity of Extracellular Vesicle Production From the Plasma Membrane.
AU - Bianca L Guerra
AU - Tanner Chase White
AU - Alissa Rumin
AU - Lauren Pitts
AU - Alexander Nguyen
AU - Vanshika Chauhan
AU - Dinah Loerke
AU - Ann M Wehman
PY - 2026
DA - 2026/11/05
JO - Journal of extracellular biology
DO - 10.1002/jex2.70197
UR - https://europepmc.org/article/MED/42996306
AB - Extracellular vesicle (EV) biogenesis is linked to the disruption of lipid bilayer asymmetry during diverse processes from coagulation to programmed cell death. However, the specific contributions of lipid scramblase proteins to the regulation of EV production and the potential redundancy of diverse families of scramblases are not well understood. We developed a semi-automated image analysis pipeline that accurately quantifies fluorescently labelled EVs on the cell surface of live Caenorhabditis elegans embryos. Using this method in a sensitized background with enhanced EV production, we performed an RNA interference (RNAi) screen targeting a subset of plasma membrane and intracellular scramblases. We found that knockdown of most scramblase genes we tested resulted in a minor but significant reduction in the number of EVs. We confirmed these significant decreases in EV abundance with anoh-2 and atg-9 scramblase mutants. However, loss-of-function egli-1 or scrm-3 scramblase mutants displayed no significant change in EV production, unlike egli-1 or scrm-3 knockdown. We also observed no effect on EV production in ced-7 ABC transporter mutants. The discrepancy between the phenotypes of acute scramblase knockdown and chronic loss suggested the presence of compensatory mechanisms that maintain EV homeostasis in response to the permanent absence of a scramblase. RNA sequencing revealed changes in expression levels for conserved sets of genes in egli-1 and scrm-3 mutants. Together, our data suggest that multiple scramblases involved in diverse pathways regulate EV budding, potentially responding to distinct physiological or metabolic cues.
SN - 2768-2811
ER -