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The in vivo effects of electroporation on the endothelial cells of blood vessels

STSM by Bostjan Markelc, PhD student

Period of mission: from 15/10/2012 to 30/11/2012

Home institution: Institute of Oncology, Dept. of Experimental Oncology, Ljubljana, Slovenia

Host institution: Institute of Pharmacology and Structural Biology – CNRS, Dept. of Cellular Biophysics, Toulouse, France – Dr. Muriel Golzio

In the STSM we addressed the blood flow modifying effects of electroporation/electropermeabilization (EP) which have been observed in vivo in muscles and tumors. Recently it has been shown that a transient constriction of normal blood vessels occurs immediately after EP, which is accompanied by an increase in their permeability. The aim of the STSM was to determine the effects of EP on the integrity of the blood vessels wall and the endothelial cells in vivo. In the project we used C57Bl/6 mice strain in which we surgically fitted the dorsal window chamber. For visualization of endothelial cells in blood vessels we used Alexa Fluor® 555 labeled CD31 antibodies and to visualize the increased permeability of blood vessels we used fluorescently labelled dextrans (FD). Mice were subjected to EP (8 pulses, 520 V/4 mm, 100 μs, 1 Hz). Image acquisition was performed with a multiphoton microscope and the subsequent image processing and analysis by Imaris software. The results show that the labeling of the endothelial cells with fluorescent antibodies enables direct observation of the effects of EP at the level of endothelial cells. We determined that after EP, there is an increase of permeability in blood vessels for FD and that this increase is due to the transient disruption of the cell-to-cell junctions in the endothelial cells of blood vessels, which however started to recover already within the first hour after EP. This was in vivo confirmation of the observed in vitro effects of EP.

This report is also featured in the February 2013 newsletter available here.

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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