Stable hybrid myotubes: a new model for studying re-expression of enzymatic activities in vitro.
Heterokaryons represent a stable and reproducible model system for the study of biochemical and molecular aspects responsible for muscle gene activation. Previous experiments have used this fusion system to demonstrate human gene activation in hybrids formed between human and non-human cells. The aim of this research was to apply this experimental model to the correction of a cytoplasmic activity, namely glucose-6-phosphate dehydrogenase (G6PD), in vitro, in hybrid myotubes formed between G6PD-negative and positive myoblasts. Different identification methods were used (Hoechst stain and Fluorescent Latex Microspheres, FLMs) to identify hybrid myotubes formed. We demonstrated the restoration of G6PD activity in all hybrid myotubes formed; we then tried to elucidate the mechanisms underlying the restoration of this specific activity and apply the results obtained to the understanding of more complex mechanisms involved in muscle gene activation.
Meola
G
G
Divisione di Neurologia, Ospedale San Donato, San Donato Milanese.
Sansone
V
V
Rotondo
G
G
Radice
S
S
Bottiroli
G
G
Scarlato
G
G
D000818:Animals; D002459:Cell Fusion; D002478:Cells, Cultured; D003593:Cytoplasm / Q000201:enzymology; D005453:Fluorescence; D015870:Gene Expression; D005954:Glucosephosphate Dehydrogenase / Q000096:biosynthesis; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000235:genetics; D006651:Histocytochemistry; D006801:Humans; D006822:Hybrid Cells; D007840:Latex; D051379:Mice; D008863:Microspheres; D008870:Microtubules / Q000201:enzymology*; D008954:Models, Biological; D009132:Muscles / Q000201:enzymology* / Q000648:ultrastructure; D013194:Staining and Labeling
D007840:Latex; D005954:Glucosephosphate Dehydrogenase