Muscle transfection and permeabilization induced by electrotransfer or pluronic® L64: Paired study by optical imaging and MRI

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• 作者：Michel F. Bureau ; Lauriane Jugé ; Johanne Seguin ; Marie-Noë ; lle Rager ; Daniel Scherman ; Nathalie Mignet
• 关键词：Block copolymer ; Permeability ; Transfection ; Electroporation ; MRI ; Optical imaging
• 刊名：Biochimica et Biophysica Acta (BBA)/General Subjects
• 出版年：2010
• 出版时间：May 2010
• 年：2010
• 卷：1800
• 期：5
• 页码：537-543
• 全文大小：1009 K

文摘

Background

Muscle transfection by electrotranfer is an efficient currently used procedure. Recently, the block copolymer pluronic L64 has been reported to improve muscle transfection. Both procedures are known to permeabilize muscle fibres. Relation between muscle transfection and permeabilization by electrotransfer and L64 was investigated herein.

Methods

Muscle transfection was evaluated by optical detection of the luciferase reporter gene activity. Muscle permeabilization was evaluated by the uptake of the T1 contrast agent gadolinium-Dotarem (Gd-DOTA) using Magnetic resonance imaging (MRI). Histological examination of muscle sections was also performed.

Results

Electrotransfer and L64 (at a 0.25 % concentration) similarly improved muscle transfection, although the interindividual variability was higher for pluronic. On the same animals, the permeabilized volume to the Gd-DOTA was significantly increased after electrotransfer, and L64 from 0.1 % to 1 % . The concentration of the Gd-DOTA in the permeabilized volume was significantly increased after electrotransfer and L64 at 0.5 % and 1 % . By histological observation, the inflammation was maximum at day 3 after electrotransfer or L64 injection, and mostly reversed after 7 days. The permeabilized volume and the transfection level correlated for the set of all the conditions tested. However, no significant correlation was observed between Gd-DOTA concentration and transfection.

General significance

It is possible to use successively on the same animals MRI and optical imaging for paired studies of muscle transfection and permeabilization. Permeabilization is possibly not related to gene transfer but it indicates membrane modification related to transfection by the electrotransfer or co-injection of DNA with the L64.