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Hypercatabolic states caused by injury are characterised by a loss of lean body mass. This form of malnutrition is associated with an increased morbidity and mortality. The muscle content of Insulin-like Growth Factor-I (IGF-I) has been reported to decrease in response to injury. Considering the anabolic and the anti-catabolic properties of this growth factor, these date strongly suggest that low muscle IGF-I may contribute to the protein hypercatabolism responsible for the loss of lean body mass.
In this study, we have developed an in vivo electroporation transfection method of the IGF-I gene in rat tibialis anterior muscle. The ultimate goal is to develop this new therapeutic approach for the prevention or treatment of loss of lean body mass.
The first part of this study has consisted in the investigation of optimal electroporation conditions. We demonstrated that electroporation is an efficient transfection method. Combining electroporation and plasmid multi-sites injections, we showed a 200-fold increase of a reporter gene expression (β-galactosidase) in the tibialis anterior muscle of rats three days after administration. This over-expression was stable up to seven days after transfection.
in the second part of this work, we have constructed two specific IGF-I plasmids. The first one encoding IGF-I and the second one encoding the IGF-I and a fluorescent GFP20 protein used as a transfection dye. A three-fold increase in IGF-I peptide muscular content was observed after in vivo electroporation of the first plasmid into tibialis anterior muscle of hypophysectomised rats. The second plasmid was transfected in vitro into mouse muscular cells (C2C12) via liposome. A specific fluorescent signal was observed in 2.5% the cells, illustrating the success of the transfection.
In conclusion, in this original work, we have demonstrated the feasibility and the efficiency of the in vivo gene transfection performed by electroporation of naked cDNA into muscle. Moreover, our combined in vitro and in vivo experimental results indicated that both plasmids are functional in an eukaryote environment Les situations hypercataboliques causées par une agression sont caractérisées par un état de déplétion affectant surtout la masse maigre. Cette forme de dénutrition est associée à une augmentation de morbidité et de mortalité.
Une diminution du contenu musculaire en « Insulin-like Growth Factor-I » a pu être mise en évidence dans des modèles hypercataboliques. Compte tenu des propriétés anaboliques et anticataboliques de ce facteur de croissance, ces données suggèrent que la diminution d’IGF-I musculaire pourrait contribuer à l’hypercatabolisme protéique responsable de la perte de masse maigre dans ces situations.
Dans ce travail, nous avons cherché à mettre au point une méthode de transfection par électroporation du gène de l’IGF-I dans le muscle tibial antérieur de rat.
La première partie de ce travail a été consacrée à la mise au point des conditions de l’électroporation. Nous montrons que l’électroporation est une méthode de transfection efficace. Combinée avec de multiples injections de plasmide, elle augmente l’expression du gène témoin, la β-galactosidase, de ceux cents fois au niveau du muscle tibial antérieur de rat. Cette expression est stable au moins sept jours.
Lors de ce travail, nous avons également construit deux vecteurs IGF-I, le premier codant pour le gène de l’IGF-I et le deuxième codant pour le gène et pour une protéine fluorescente la GFP20. L’électroporation du premier vecteur dans le muscle de rats hypophysectomisés augmente de trois fois le contenu musculaire d’IGF-I. le deuxième vecteur a été transfecté par des liposomes dans des cellules musculaires C2C12 en culture. Le succès de la transfection est illustré par la mise en évidence d’une fluorescence intracellulaire dans 2.5% des cellules.
Nous avons démontré la faisabilité et l’efficacité du transfert de gène par une association de multiples injections d’ADN nu combinées avec l’électroporation dans des cellules musculaires in vivo. Les expériences in vitro et in vivo ont montré que les vecteurs construits s’expriment correctement dans un environnement eucaryote

Insulin-Like Growth Factor I --- Muscle, Skeletal --- Electroporation --- Transfection

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Biological techniques --- Molecular biology --- Genetic transformation --- Gene expression --- Gene Expression Regulation --- Transfection --- Laboratory manuals --- Regulation --- Gene Expression Regulation. --- Transfection. --- 575.117 --- 577.216 --- 57.088.6 --- -Genetic transformation --- -#WSCH:WBIO --- Gene transfer --- Transformation (Genetics) --- Genetic recombination --- Microbial genetics --- Nucleic acids --- Genes --- Genetic regulation --- Transfections --- Transformation, Bacterial --- Transformation, Genetic --- Expression Regulation, Gene --- Regulation, Gene Action --- Regulation, Gene Expression --- Gene Action Regulation --- Regulation of Gene Expression --- RNAi Therapeutics --- Gene Regulatory Networks --- Expressivity and penetrance of genes --- Transfer of inheritance information. Transport of messenger RNA in the cell --- Methods and techniques for studying metabolism and biotransformation.Radioisotopes methods and techniques. --- -Laboratory manuals --- Expression --- Laboratory manuals. --- laboratory manuals. --- 57.088.6 Methods and techniques for studying metabolism and biotransformation.Radioisotopes methods and techniques. --- 577.216 Transfer of inheritance information. Transport of messenger RNA in the cell --- 575.117 Expressivity and penetrance of genes --- laboratory manuals --- #WSCH:WBIO --- Regulation&delete& --- Methods and techniques for studying metabolism and biotransformation.Radioisotopes methods and techniques --- Genetic transformation - Laboratory manuals --- Gene expression - Regulation - Laboratory manuals --- Gene Expression Regulation - laboratory manuals --- Transfection - laboratory manuals --- GENE EXPRESSION REGULATION --- TRANSFECTION --- LABORATORY MANUALS

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In 1995, John Maynard Smith and Eörs Szathmáry published their influential book 'The Major Transitions in Evolution'. In this volume, scholars reconsider and extend the earlier book's themes in light of recent developments in evolutionary biology.

Evolution (Biology) --- Biodiversity. --- Genetic transformation. --- Population genetics. --- Maynard Smith, John, --- Szathmáry, Eörs. --- Gene transfer --- Transformation (Genetics) --- Biological diversification --- Biological diversity --- Biotic diversity --- Diversification, Biological --- Diversity, Biological --- Animal evolution --- Animals --- Biological evolution --- Darwinism --- Evolutionary biology --- Evolutionary science --- Origin of species --- Evolution --- Genetics --- Heredity --- Genetic recombination --- Microbial genetics --- Nucleic acids --- Transfection --- Biology --- Biocomplexity --- Ecological heterogeneity --- Numbers of species --- Biological fitness --- Homoplasy --- Natural selection --- Phylogeny --- BIOMEDICAL SCIENCES/Evolution --- Évolution (biologie) --- Biodiversité. --- Transformation génétique. --- Génétique des populations. --- Evolution. --- Natural selection. --- Acqui 2006 --- Genetic transformation --- POPULATION GENETICS --- Évolution (biologie) --- Biodiversité. --- Transformation génétique. --- Génétique des populations.