Cytoskeletal involvement in the cellular trafficking of plasmid/PEI derivative complexes

Abstract : We have studied the cytoskeletal involvement in the cellular trafficking of complexes made with plasmid/PEI or plasmid/lactosylated PEI in cystic fibrosis airway epithelial cells (SigmaCFTE29o- cells). Complexes were incubated in the presence of cytoskeletal inhibitors, and the number of transfected cells was determined by flow cytometry. Complexes were also generated with fluorescein-labeled PEI derivatives and the cell fluorescence intensity was determined by flow cytometry. In the presence of cytochalasin D to depolymerize actin filaments or nocodazole to disrupt microtubules, gene transfer efficiency with both PEI derivatives was decreased by 90%. The uptake of fluoresceinylated complexes studied by flow cytometry was decreased by 50% in the presence of cytochalasin D for both types of complexes (p<0.005) and unchanged in the presence of nocodazole. When cytoskeletal inhibitors were added to the cell culture after the complex uptake had occurred, gene transfer efficiency was decreased by 75% and 50% in the presence of nocodazole and cytochalasin D, respectively. Upon nocodazole-microtubule network disruption, the lysosomal localization of complexes was reduced, as assessed by confocal microscopy. Our results show a major cytoskeletal involvement in the cellular trafficking of complexes made with both PEI derivatives: actin filaments mainly in complex uptake, and microtubules in the trafficking of complexes towards the nucleus, probably through guided transport of complex-containing endosomal vesicles.
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Journal articles
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https://hal.archives-ouvertes.fr/hal-00166244
Contributor : Isabelle Frapart <>
Submitted on : Thursday, August 2, 2007 - 12:33:49 PM
Last modification on : Thursday, April 11, 2019 - 4:02:57 PM

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Stéphanie Grosse, Yolande Aron, Guiti Thévenot, Michel Monsigny, Isabelle Fajac. Cytoskeletal involvement in the cellular trafficking of plasmid/PEI derivative complexes. Journal of Controlled Release, Elsevier, 2007, 122 (1), pp.111-117. ⟨10.1016/j.jconrel.2007.06.015⟩. ⟨hal-00166244⟩

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