NDLI: Involvement of cytoskeleton in sonoporation and drug delivery

Content Provider	IEEE Xplore Digital Library
Author	Zeghimi, A. Escoffre, J.-M. Bouakaz, A.
Copyright Year	2014
Description	Author affiliation: Inserm, Univ. Francois-Rabelais de Tours, Tours, France (Zeghimi, A.; Escoffre, J.-M.; Bouakaz, A.)
Abstract	The mechanisms by which sonoporation increases the native plasma membrane permeability are still unknown but various hypotheses have been suggested including pore formation and endocytosis. We have shown recently that caveolae-mediated endocytosis plays a major role during sonoporation. In addition, some studies have reported the clear participation of actin in mammalian cell endocytosis. Thus, this study aims to investigate the effect of sonoporation on actin microfilaments and microtubules and to identify the role of both cytoskeletons on sonoporation-mediated membrane permeabilization. Adherent U-87 MG cells were insonated at 1MHz, 1 $W/cm^{2},$ 20% duty cycle for 60 s, in the presence of $BR14^{®}$ microbubbles. $SYTOX^{®}$ Green was used to assess the membrane permeabilization, by flow cytometry. The cells were incubated with phalloidin-TRITC to stain actin microfilaments and tubulin antibody Alexa $Fluor^{®}$ 555 to stain tubulin. The ultrastructural changes of plasma membrane were monitored by scanning electron microscopy. To inhibit the polymerization of actin and tubulin cytoskeleton, the cells were treated with cytochalasin D (cytoD) and nocodazole (Noco), respectively. Immunofluorescence results show alteration of actin and tubulin cytoskeleton, immediately after sonoporation while control cells present a filamentous cytoskeleton with polygonal shape. However, the disorganization of the cytoskeleton network is reversible since 60 min post-sonoporation, only few cells show a tubulin (8%) and actin (25%) cytoskeleton disruption. Moreover based on SEM study reveals that the treatment of the cells with both cytoD and Noco induced a strong decrease in the number of TPS (transient and permeant structures): 98.5 ± 0.2% and 96 ± 0.6%, respectively. Moreover, flow cytometry results showed that cytoD and Noco lead to a decrease in the membrane permeabilization rate: 58% and 87%, respectively. In conclusion, this study demonstrates the transient alteration of actin and tubulin cytoskeleton following sonoporation. It suggests that cytoskeleton plays a role during sonoporation, as cytoskeleton inhibitors provoke a decrease in the cell permeabilization rate. Its implication could occur during both the entry and transport of endocytosed molecules.
Starting Page	850
Ending Page	853
File Size	721671
Page Count	4
File Format	PDF
e-ISBN	9781479970490
DOI	10.1109/ULTSYM.2014.0209
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2014-09-03
Publisher Place	USA
Access Restriction	Subscribed
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	Polymers Plasmas Monitoring Biomedical monitoring Immune system Permeability Scacnning electron microscopy Sonoporation Ultrasound Microbubbles Cytoskeleton Immunofluorescence
Content Type	Text
Resource Type	Article

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