Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids.
Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids.
复制标题
DOI：
10.1038/srep31172
复制
发表时间：
2016-08-19
影响因子：
4.6
通讯作者：
Tamai Y
中科院分区：
文献类型：
--
作者：
Yoshimura A;Kawamata M;Yoshioka Y;Katsuda T;Kikuchi H;Nagai Y;Adachi N;Numakawa T;Kunugi H;Ochiya T;Tamai Y
细胞外囊泡在生物分子在细胞间的传递中起着重要的作用。为了阐明EV在体内的细胞间转移命运，我们使用绿色荧光蛋白（GFP）标记的人CD 63产生了一种新的转基因（Tg）大鼠模型。CD 63蛋白通过运输到晚期内体而高度富集在EV膜上，并且通常用作EV标志物。人CD 63-GFP受CAG启动子控制的新Tg大鼠系在各种身体组织中表现出GFP的高表达。在从Tg大鼠的三种体液（血清、母乳和羊水）分离的EV上检测到外源性人CD 63-GFP。体外培养允许将血清来源的CD 63-GFP EV转移到受体大鼠胚胎成纤维细胞中，其中EV定位于内吞细胞器中。这些结果表明，这种Tg大鼠模型应该提供重要的信息，了解细胞间转移和/或母子转移的EV在体内。
Extracellular vesicles (EVs) play an important role in the transfer of biomolecules between cells. To elucidate the intercellular transfer fate of EVs in vivo, we generated a new transgenic (Tg) rat model using green fluorescent protein (GFP)-tagged human CD63. CD63 protein is highly enriched on EV membranes via trafficking into late endosomes and is often used as an EV marker. The new Tg rat line in which human CD63-GFP is under control of the CAG promoter exhibited high expression of GFP in various body tissues. Exogenous human CD63-GFP was detected on EVs isolated from three body fluids of the Tg rats: blood serum, breast milk and amniotic fluid. In vitro culture allowed transfer of serum-derived CD63-GFP EVs into recipient rat embryonic fibroblasts, where the EVs localized in endocytic organelles. These results suggested that this Tg rat model should provide significant information for understanding the intercellular transfer and/or mother-child transfer of EVs in vivo.