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A transgenic inducible GFP extracellular-vesicle reporter (TIGER) mouse illuminates neonatal cortical astrocytes as a source of immunomodulatory extracellular vesicles
Abstract
Extracellular vesicles (EVs) are cellular derived particles found throughout the body in nearly all tissues and bodily fluids. EVs contain biological molecules including small RNAs and protein. EVs are proposed to be transferred between cells, notably, cells of the immune system. Tools that allow for in vivo EV labeling while retaining the ability to resolve cellular sources and timing of release are required for a full understanding of EV functions. Fluorescent EV fusion proteins are useful for the study of EV biogenesis, release, and identification of EV cellular recipients. Among the most plentiful and frequently identified EV proteins is CD9, a tetraspanin protein. A transgenic mouse containing a CRE-recombinase inducible CAG promoter driven CD9 protein fused to Turbo-GFP derived from the copepod Pontellina plumata was generated as an EV reporter. The transgenic inducible GFP EV reporter (TIGER) mouse was electroporated with CAG-CRE plasmids or crossed with tamoxifen inducible CAG-CRE-ER T2 or nestin-CRE-ER T2 mice. CD9-GFP labeled cells included glutamine synthetase and glial fibrillary acidic protein positive astrocytes. Cortical astrocytes released ~136 nm EVs that contained CD9. Intraventricular injected EVs were taken up by CD11b/IBA1 positive microglia surrounding the lateral ventricles. Neonatal electroporation and shRNA mediated knockdown of Rab27a in dorsal subventricular zone NSCs and astrocytes increased the number of CD11b/IBA1 positive rounded microglia. Neonatal astrocyte EVs had a unique small RNA signature comprised of morphogenic miRNAs that induce microglia cytokine release. The results from this study demonstrate that inducible CD9-GFP mice will provide the EV community with a tool that allows for EV labeling in a cell-type specific manner while simultaneously allowing in vivo experimentation and provides evidence that EVs are required immunomodulators of the developing nervous system.
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“Thus, identifying miRNAs that are exclusively expressed in sEVs from distinct donor cells may enable us to define the cellular origin of the circulating sEVs [50, 51].”
From Differential RNA packaging into small extracellular vesicles by neurons and astrocytes (2021) · cites this paper as [51]
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Astrocyte-derived extracellular vesicles: A double-edged sword in central nervous system disorders
Zhao, S., Sheng, S., Wang, Y., Ding, L., Xu, X., Xia, X., & Zheng, J. C. (2021) · Neuroscience & Biobehavioral Reviews · doi:10.1016/j.neubiorev.2021.02.027 · Cited by 90
Differential RNA packaging into small extracellular vesicles by neurons and astrocytes
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Metadata
- Authors
- Neckles, V. N., Morton, M. C., Holmberg, J. C., Sokolov, A. M., Nottoli, T., Liu, D., & Feliciano, D. M.
- Venue
- Scientific Reports
- Published
- 2019-02-28
- Volume
- 9
- Issue
- 1
- ISSN
- 2045-2322
- PMC
- PMC6395689
- Cited by
- 55
- References
- 6
- Language
- en
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