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Staining of cell-to-cell junctions after EP - in vitro study

STSM by Tanja Dolinšek, PhD Student

Period of mission: from 09/09/2013 to 04/10/2013

Home institution: Institute of Oncology, Department of Experimental Oncology, Ljubljana, Slovenia

Host institution: Institut of Pharmacology and Structural Biology (IPBS) - CNRS, Toulouse, France

It has been reported that a transient constriction of normal blood vessels occurs immediately after application of electric field pulses (EP). This phenomenon is accompanied by an increase in their permeability. In a previous STSM, Bostjan Markelc observed that the increase of the permeability of blood vessels was most probably due to the transient disruption of the cell-to-cell junctions in endothelial cells. Namely, the fluorescence of AlexaFluor 555 labeled antibody against CD31 became blurred and dispersed after application of EP, suggesting a direct effect on cell-to-cell junctions. However, whether the increased permeability was due to trans- or para- cellular transport was not known. Therefore, the aim of my STSM was to confirm, complement and further elaborate these results obtained in vivo in mice by using an in vitro setting. For that purpose, I stained mouse endothelial cells with cell junctions’ specific antibodies (anti CD31 and VE-cadherin) and observed the changes in cell-to-cell junctions before and at different times after EP. A very specific staining of cell-to-cell junctions has been obtained with VE-cadherin antibodies. However the staining for CD31 was more dispersed and not completely localized to cell junctions. I observed that in the cell monolayer submitted to EP (600 V/cm, 8 pulses, 100 µs, 1 Hz, plate electrodes 7.3 mm), the intensity of staining of cell junctions was immediately decreased and started to recover 10 min after EP. These data therefore confirms the results obtained in vivo, showing that the permeability of blood vessels is due to the transient disruption of cell-to-cell junctions of endothelial cells.

Poster presentation(s) originating from this mission

Event Annual Congress of the French Society of Cell and Gene Therapy (SFTCG)
Location Toulouse/France
Period

24/03 - 26/03/2014

Authors Bostjan Markelc, Elisabeth Bellard, Tanja Dolinsek, Sandrine Pelofy, Gregor Sersa, Marie-Pierre Rols, Justin Teissie, Maja Cemazar, Muriel Golzio
Title Determining the effects of electric pulses on endothelial cells of normal blood vessels with longitudinal biphoton microscopy imaging

 


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