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Induced endocytosis in human fibroblasts by electrical fields
M Glogauer1, W Lee, C A McCulloch
1Faculty of Dentistry, University of Toronto, Ontario, Canada.
Experimental Cell Research
|September 1, 1993
Summary
Electroporation, a method for cell molecule delivery, increases cell uptake of substances via endocytosis. This actin-dependent process enhances the internalization of membrane components and probe entry into cells.
Area of Science:
- Cell Biology
- Biophysics
- Molecular Medicine
Background:
- Electroporation facilitates cellular uptake of molecules by creating temporary pores.
- Electrical fields during electroporation may disrupt membrane functions like endocytosis.
Purpose of the Study:
- To investigate the impact of electroporation on endocytosis in human gingival fibroblasts.
- To determine if electroporation affects the internalization of various membrane components and probes.
Main Methods:
- Human gingival fibroblasts were electroporated and allowed to recover.
- Cells were incubated with fluorescent probes (FITC-BSA, FITC-Concanavalin A, lucifer yellow) and analyzed via flow cytometry and confocal microscopy.
- Actin polymerization was inhibited using cytochalasin D; enzyme treatments (endoglycosidase F/H) and specific inhibitors (alpha-methyl D-mannoside) were used to confirm probe specificity and internalization pathways.
Main Results:
- Electroporation significantly increased the internalization of FITC-BSA, lucifer yellow, and surface glycoproteins in a time-dependent manner.
- Cytochalasin D pretreatment abrogated the electroporation-induced increase in FITC-BSA internalization, indicating an actin-dependent mechanism.
- Electroporation led to increased acidification within endosomal compartments, suggesting faster entry of membrane lipids.
Conclusions:
- Electroporation induces an actin-dependent enhancement of endocytosis in fibroblasts.
- The electrical fields used in electroporation promote the internalization of diverse membrane components.
- Electroporation accelerates the rate at which probes enter acidifying endosomal compartments.