Early effects of AZT on mitochondrial functions in the absence of mitochondrial DNA depletion in rat myotubes.
Zidovudine (AZT) is a potent inhibitor of human immunodeficiency virus (HIV) replication. In humans, as well as in animal models, long-term treatment with AZT induces a severe myopathy characterised by structural and functional alterations of mitochondria associated with depletion of mitochondrial DNA (mtDNA). In the present work, we compared the effects induced by AZT on mitochondria upon short- or long-term treatments of cultured rat myotubes. Morphological alterations were investigated by electron microscopy, and mtDNA depletion and deletions were analysed by Southern blot. Mitochondrial membrane potential was determined after JC-1 staining by laser-scanning confocal microscopy in whole cells, and by flow cytometry in isolated muscle mitochondria. We found that the early effects of AZT on mitochondrial functions were a marked, yet reversible reduction in mitochondrial membrane potential, in the absence of any effect on mtDNA. The long-term treatment, in addition to mitochondrial membrane potential alterations, induced morphological changes in mitochondria, and a remarkable reduction in the amount of mtDNA, without any significant evidence of mtDNA deletions. In both treatments, a block of the spontaneous contraction of myotubes was observed. To study in more detail the early effects induced by AZT, the ability of the drug to interact with cardiolipin, an important component of internal mitochondrial membrane, was investigated by atomic force microscopy (AFM) in an artificial membrane model system. The results suggest that the primary effects of AZT may be related to a physical interference with the membrane structure leading to a consequent modification of its physical characteristics.
Cazzalini
O
O
Dipartimento di Medicina sperimentale, sez. Patologia generale C.Golgi , P.zza Botta 10; I-27100, Pavia, Italy. ornella@manuzio.unipv.it
Lazzè
M C
MC
Iamele
L
L
Stivala
L A
LA
Bianchi
L
L
Vaghi
P
P
Cornaglia
A
A
Calligaro
A
A
Curti
D
D
Alessandrini
A
A
Prosperi
E
E
Vannini
V
V
D000818:Animals; D002478:Cells, Cultured; D004272:DNA, Mitochondrial / Q000187:drug effects*; D006651:Histocytochemistry; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D018625:Microscopy, Atomic Force; D008928:Mitochondria / Q000187:drug effects* / Q000235:genetics / Q000502:physiology / Q000648:ultrastructure; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D015215:Zidovudine / Q000494:pharmacology*
D004272:DNA, Mitochondrial; D015215:Zidovudine