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Ultrastructural Analysis of Vesicular Transport in Electrotransfection

Published online by Cambridge University Press:  18 October 2018

Liangli Wang ,
Sara E. Miller  and
Fan Yuan
Liangli Wang
Affiliation:
Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA
Sara E. Miller*
Affiliation:
Department of Pathology, Duke University Medical School, Durham, NC 27710, USA
Fan Yuan*
Affiliation:
Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA
*
*Authors for correspondence: Sara E. Miller, E-mail: [email protected]; Fan Yuan, E-mail: [email protected]
*Authors for correspondence: Sara E. Miller, E-mail: [email protected]; Fan Yuan, E-mail: [email protected]
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Abstract

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/glycosylphosphatidylinositol-anchored protein-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.

Keywords

electrotransfectionelectroporationintracellular traffickingclathrin-mediated endocytosismacropinocytosis
Type
Biological Science Applications
Information
Microscopy and Microanalysis , Volume 24 , Issue 5 , October 2018 , pp. 553 - 563
Copyright
© Microscopy Society of America 2018 

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