A transgenic inducible GFP extracellular-vesicle reporter (TIGER) mouse illuminates neonatal cortical astrocytes as a source of immunomodulatory extracellular vesicles

Scientific Reports
Victoria N NecklesDavid M Feliciano

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-ERT2 or nestin-CRE-ERT2 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...Continue Reading

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Citations

Jun 17, 2020·Genesis : the Journal of Genetics and Development·James V McCannAndrew C Dudley
May 23, 2020·Scientific Reports·Joana Maria Ramis
Jun 17, 2020·Zoological Letters·Marie-Lyne MacelSalvatore D'Aniello
Feb 25, 2021·Neuroscience and Biobehavioral Reviews·Shu ZhaoJialin C Zheng
Apr 4, 2021·Developmental Cell·Bong Hwan SungAlissa M Weaver
Jun 10, 2021·Journal of Tissue Engineering·Jung-Hwan LeeHae-Won Kim
Jun 22, 2021·Cell and Tissue Research·Victoria N Neckles, David M Feliciano
Jul 12, 2021·Cell Communication and Signaling : CCS·Xuan LuoSeena K Ajit
Aug 12, 2021·FASEB BioAdvances·Andrea SchnatzEva-Maria Krämer-Albers

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Methods Mentioned

BETA
transgenic
PCR
confocal microscopy
RNA assay
Hi-Seq
transfection
protein assay

Software Mentioned

Banana Slug Analytics Platform
GraphPad 7 Prism
FIJI
GraphPad Prism
Nanosight
CLARITY
GraphPad
Image J
Prism

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