Ultrastructural Analysis of Vesicular Transport in Electrotransfection.
Ultrastructural Analysis of Vesicular Transport in Electrotransfection.
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DOI：
10.1017/s143192761801509x
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发表时间：
2018-10
期刊：
影响因子：
--
通讯作者：
Yuan F
中科院分区：
文献类型：
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作者：
Wang L;Miller SE;Yuan F
来自各种研究的新证据表明，质粒DNA（pDNA）在用于电转染时通过胞吞样过程被细胞内化。为了提供该过程的形态学证据，我们使用免疫电子显微镜研究了与电转染的pDNA相关的细胞中的超微结构。结果表明，四种内吞途径参与pDNA的摄取，包括小窝和网格蛋白介导的内吞作用、巨胞饮作用和网格蛋白非依赖性载体/GPI富集的早期内体室（CLIC/GEEC）途径。其中，巨胞饮作用是具有各种pDNA摄取能力的细胞所利用的最常见途径，并且CLIC/GEEC途径主要在人脐静脉内皮细胞中观察到。从数量上看，内吞途径在易被吞噬的细胞中比在难被吞噬的细胞中更活跃。两者合计，我们的数据提供了超微结构的证据表明，内吞作用起着重要的作用，在细胞摄取和细胞内转运的电转染的pDNA。
Emerging evidence from various studies indicates that plasmid DNA (pDNA) is internalized by cells through an endocytosis-like process when it is used for electrotransfection. To provide morphological evidence of the process, we investigated ultrastructures in cells that were associated with the electrotransfected pDNA, using immunoelectron microscopy. The results demonstrate that four endocytic pathways are involved in the uptake of the pDNA, including caveolae- and clathrin-mediated endocytosis, macropinocytosis, and the clathrin- independent carrier/GPI-enriched early endosomal compartment (CLIC/GEEC) pathway. Among them, macropinocytosis is the most common pathway utilized by cells having various pDNA uptake capacities, and the CLIC/GEEC pathway is observed primarily in human umbilical vein endothelial cells. Quantitatively, the endocytic pathways are more active in easy-to-transfect cells than in hard-to-transfect ones. Taken together, our data provide ultrastructural evidence showing that endocytosis plays an important role in cellular uptake and intracellular transport of electrotransfected pDNA.