Evidence for electro-induced membrane defects assessed by lateral mobility measurement of a GPi anchored protein
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Electrotransfer is a method by which molecules can be introduced into living cells via plasma membrane electropermeabilization. Here, we show that electropermeabilization affects the lateral mobility of Rae-1, a GPi anchored protein. Our results suggest that 10–20 % of the membrane surface is occupied by defects or pores and that these structures propagate rapidly (\(<\)1 min) over the cell surface. Electrotransfer of plasmid DNA (pDNA) also affects the lateral mobility of Rae-1. Furthermore, we clearly show that, once inserted into the plasma membrane, pDNA is completely immobile and excludes Rae-1; this indicates that the pDNA molecules are tightly packed together to form aggregates occupying at least the outer leaflet of the plasma membrane.
KeywordsElectropermeabilization GPi anchored protein pDNA FRAP Lateral mobility Pores
This work has been performed in collaboration with the “Toulouse Réseau Imagerie” core IPBS facility (Genotoul, Toulouse, France), which is supported by the Association Recherche Cancer, Region Midi Pyrenees, the European union (FEDER) and Grand Toulouse cluster. This research project was conducted in the scope of EBAM European Associated Laboratory and of the COST Action TD1104. JM Escoffre was the recipient of an allocation de recherche du Ministère de l’Enseignement Supérieur et de la Recherche. The authors are grateful to Dr. M. Golzio and Dr. E. Bellard for critical reading of the manuscript and Dr. T. Melvin for English rewriting. C. Favard is a membership of CNRS consortii GDR2588 “MIV” and GDR3070 “CellTiss”.
- Chang D, Chassy B, Saunders J, Sowers A (1992) Guide to electroporation and electrofusion. San DiegoGoogle Scholar
- Escoffre JM, Rols MP (2012) Electrochemotherapy: progress and prospects. Curr Pharm Des 18(23):3406–3415Google Scholar
- Matthews C, Favard C (2007) Theory, principles and applications of fluorescent technologies in cellular biology and cancer research. Bull Cancer (French) 94(1):115–125Google Scholar
- Mir LM, Gehl J, Sersa G, Collins CG, Garbay JR, Billard V, Geertsen PF, Rudolf Z, O’Sullivan GC, Marty M (2006) Standard operating procedures of the electrochemotherapy: instructions for the use of bleomycin or cisplatin administered either systemically or locally and electric pulses delivered by the Cliniporator (TM) by means of invasive or non-invasive electrodes. EJC Suppl 4(11):14–25CrossRefGoogle Scholar
- Neumann E, Sowers A, Jordan C (1989) Electroporation and electrofusion in cell biology, Plenum, New YorkGoogle Scholar
- Romeo S, Wu YH, Levine ZA, Gundersen MA (1828) Vernier PT (2013) Water influx and cell swelling after nanosecond electropermeabilization. Biochim Biophys Acta 8:1715–1722Google Scholar
- Weiland O, AhlTn G, Diepolder H, Jung MC, Levander S, Fons M, Mathiesen I, Sardesai NY, Vahlne A, Frelin L, SSllberg M (2013) Therapeutic dna vaccination using in vivo electroporation followed by standard of care therapy in patients with genotype 1 chronic hepatitis C. Mol Ther 21(9):1796–1805PubMedCentralPubMedCrossRefGoogle Scholar