Exomap1 mouse: A transgenic model for in vivo studies of exosome biology

Francis K. Fordjour , Sarah Abuelreich , Xiaoman Hong , Emeli Chatterjee , Valeria Lallai , Martin Ng , Andras Saftics , Fengyan Deng , Natacha Carnel-Amar , Hiroaki Wakimoto , Kazuhide Shimizu , Malia Bautista , Tuan Anh Phu , Ngan K. Vu , Paige C. Geiger , Robert L. Raffai , Christie D. Fowler , Saumya Das , Lane K. Christenson , Tijana Jovanovic-Talisman , Stephen J. Gould
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Abstract

Exosomes are small extracellular vesicles (sEVs) of ∼30–150 nm in diameter that are enriched in exosome marker proteins and play important roles in health and disease. To address large unanswered questions regarding exosome biology in vivo, we created the Exomap1 transgenic mouse, which in response to Cre recombinase expresses the most highly enriched exosomal marker protein known, human CD81, fused to mNeonGreen (HsCD81mNG), and prior to Cre expresses a mitochondrial red fluorescent protein. Validation of the exomap1 mouse with eight distinct Cre drivers demonstrated that HsCD81mNG was expressed only in response to Cre, that murine cells correctly localized HsCD81mNG to the plasma membrane, and that this led to the secretion of HsCD81mNG in EVs that had the size (∼70–80 nm), topology, and composition of exosomes. Furthermore, cell type-specific activation of the exomap1 transgene allowed us to use quantitative single molecule localization microscopy to calculate the cell type-specific contribution to biofluid exosome populations. Specifically, we show that neurons contribute ∼1% to plasma and cerebrospinal fluid exosome populations whereas hepatocytes contribute ∼15% to plasma exosome populations, numbers that reflect the known vascular permeabilities of brain and liver. These observations validate the use of Exomap1 mouse models for in vivo studies of exosome biology.

Exomap1小鼠:外泌体生物学体内研究的转基因模型
外泌体是直径约30 - 150nm的小细胞外囊泡(sev),富含外泌体标记蛋白,在健康和疾病中发挥重要作用。为了解决关于体内外泌体生物学的大量悬而未决的问题,我们创建了Exomap1转基因小鼠,该小鼠在响应Cre重组酶时表达已知的最富集的外泌体标记蛋白,人CD81,与mNeonGreen融合(HsCD81mNG),并在Cre表达线粒体红色荧光蛋白之前表达。对具有8种不同Cre驱动的exomap1小鼠的验证表明,HsCD81mNG仅在对Cre的响应中表达,小鼠细胞正确地将HsCD81mNG定位到质膜上,这导致在具有外泌体大小(~ 70-80 nm)、拓扑结构和组成的ev中分泌HsCD81mNG。此外,exomap1转基因的细胞类型特异性激活使我们能够使用定量单分子定位显微镜来计算细胞类型特异性对生物流体外泌体种群的贡献。具体来说,我们发现神经元对血浆和脑脊液外泌体的贡献约为1%,而肝细胞对血浆外泌体的贡献约为15%,这些数字反映了已知的脑和肝脏血管通透性。这些观察结果验证了Exomap1小鼠模型在体内外泌体生物学研究中的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Extracellular vesicle
Extracellular vesicle Biochemistry, Genetics and Molecular Biology (General)
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