Gene transfer in regenerating muscle

Maurizio Vitadello, Maria Vittoria Schiaffino, Anne Picard, Maurizio Scarpa, Stefano Schiaffino

Research output: Contribution to journalArticlepeer-review

Abstract

We have compared the efficiency of direct gene transfer in normal and regenerating rat skeletal muscle. Muscle necrosis and regeneration was induced by intramuscular injection of bupivacaine in the soleus muscle of adult rats. Plasmids containing β-galactosidase (β-gal) or chloramphenicol acetyltransferase (CAT) genes driven by viral promoters were injected 3 days after bupivacaine treatment into the regenerating and the contralateral uninjured muscles. Expression of CAT activity was > 80-fold higher in regenerating compared to control muscles at 7 days post-transfection, but decreased at 30 and 60 days. Southern blot analysis showed that the predominant form of CAT DNA was episomal in transfected muscles; however, CAT activity measurements performed on the same transfected muscles showed no precise correlation between enzymatic activity and amount of plasmid DNA. Expression of β-gal was detected in numerous regenerating fibers of the injured soleus muscles at 7 days post-transfection; in contrast, only rare positive fibers were found in control muscles. Focal infiltrates of mononuclear cells, which surround and invade selectively β-gal-positive fiber segments, were observed at 30 days post transfection, suggesting that immune mechanisms are implicated in the progressive loss of transgenes with time. The finding that regenerating muscle fibers display a higher efficiency of transfection, may be relevant to gene therapy of Duchenne muscular dystrophy, because regenerating fibers are numerous in the early stages of the disease.

Original languageEnglish
Pages (from-to)11-18
Number of pages8
JournalHuman Gene Therapy
Volume5
Issue number1
Publication statusPublished - Jan 1994

ASJC Scopus subject areas

  • Genetics

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