PUBLICATION
In Vivo Characterization of Endogenous Cardiovascular Extracellular Vesicles in Larval and Adult Zebrafish
- Authors
- Scott, A., Sueiro Ballesteros, L., Bradshaw, M., Tsuji, C., Power, A., Lorriman, J., Love, J., Paul, D., Herman, A., Emanueli, C., Richardson, R.J.
- ID
- ZDB-PUB-210716-1
- Date
- 2021
- Source
- Arteriosclerosis, Thrombosis, and Vascular Biology 41(9): 2454-2468 (Journal)
- Registered Authors
- Richardson, Rebecca
- Keywords
- cardiovascular, exosomes, extracellular vesicle, flow cytometry, zebrafish
- MeSH Terms
-
- Larva/metabolism
- Cryoelectron Microscopy
- Endothelial Cells/metabolism
- Endothelial Cells/ultrastructure
- Recombinant Fusion Proteins/genetics
- Recombinant Fusion Proteins/metabolism
- Disease Models, Animal
- Myocytes, Cardiac/metabolism
- Myocytes, Cardiac/ultrastructure
- Cell Separation
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish/metabolism*
- Gene Expression Regulation, Developmental
- Flow Cytometry
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- Cardiovascular System/embryology
- Cardiovascular System/metabolism*
- Extracellular Vesicles/genetics
- Extracellular Vesicles/metabolism*
- Extracellular Vesicles/ultrastructure
- Time Factors
- Animals
- Luminescent Proteins/genetics
- Luminescent Proteins/metabolism
- Myocardial Infarction/genetics
- Myocardial Infarction/metabolism
- Myocardial Infarction/pathology
- Animals, Genetically Modified
- PubMed
- 34261327 Full text @ Arterio., Thromb., and Vas. Bio.
Citation
Scott, A., Sueiro Ballesteros, L., Bradshaw, M., Tsuji, C., Power, A., Lorriman, J., Love, J., Paul, D., Herman, A., Emanueli, C., Richardson, R.J. (2021) In Vivo Characterization of Endogenous Cardiovascular Extracellular Vesicles in Larval and Adult Zebrafish. Arteriosclerosis, Thrombosis, and Vascular Biology. 41(9):2454-2468.
Abstract
Objective Extracellular vesicles (EVs) facilitate molecular transport across extracellular space, allowing local and systemic signaling during homeostasis and in disease. Extensive studies have described functional roles for EV populations, including during cardiovascular disease, but the in vivo characterization of endogenously produced EVs is still in its infancy. Because of their genetic tractability and live imaging amenability, zebrafish represent an ideal but under-used model to investigate endogenous EVs. We aimed to establish a transgenic zebrafish model to allow the in vivo identification, tracking, and extraction of endogenous EVs produced by different cell types. Approach and Results: Using a membrane-tethered fluorophore reporter system, we show that EVs can be fluorescently labeled in larval and adult zebrafish and demonstrate that multiple cell types including endothelial cells and cardiomyocytes actively produce EVs in vivo. Cell-type specific EVs can be tracked by high spatiotemporal resolution light-sheet live imaging and modified flow cytometry methods allow these EVs to be further evaluated. Additionally, cryo electron microscopy reveals the full morphological diversity of larval and adult EVs. Importantly, we demonstrate the utility of this model by showing that different cell types exchange EVs in the adult heart and that ischemic injury models dynamically alter EV production.
Conclusions We describe a powerful in vivo zebrafish model for the investigation of endogenous EVs in all aspects of cardiovascular biology and pathology. A cell membrane fluorophore labeling approach allows cell-type specific tracing of EV origin without bias toward the expression of individual protein markers and will allow detailed future examination of their function.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping