UCP2 muscle gene transfer modifies mitochondrial membrane potential.
OBJECTIVE
The aim of this work was to evaluate the effect of uncoupling protein 2 (UCP2) muscle gene transfer on mitochondrial activity.


METHODS
Five week-old male Wistar rats received an intramuscular injection of plasmid pXU1 containing UCP2 cDNA in the right tibialis anterior muscles. Left tibialis anterior muscles were injected with vehicle as control. Ten days after DNA injection, tibialis anterior muscles were dissected and muscle mitochondria isolated and analyzed.


RESULTS
There were two mitochondrial populations in the muscle after UCP2 gene transfer, one of low fluorescence and complexity and the other, showing high fluorescence and complexity. UCP2 gene transfer resulted in a 3.6 fold increase in muscle UCP2 protein levels compared to control muscles assessed by Western blotting. Furthermore, a significant reduction in mitochondria membrane potential assessed by spectrofluorometry and flow cytometry was observed. The mitochondria membrane potential reduction might account for a decrease in fluorescence of the low fluorescence mitochondrial subpopulation.


CONCLUSIONS
It has been demonstrated that UCP2 muscle gene transfer in vivo is associated with a lower mitochondria membrane potential. Our results suggest the potential involvement of UCP2 in uncoupling respiration. International Journal of Obesity (2001) 25, 68-74
Department of Physiology and Nutrition, University of Navarra, Pamplona, Spain.
A
A
Marti
E
E
Larrarte
F J
FJ
Novo
M
M
Garcia
J A
JA
Martinez
D000818:Animals; D015153:Blotting, Western; D018076:DNA, Complementary; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D007273:Injections, Intramuscular; D007473:Ion Channels; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D026901:Membrane Transport Proteins; D008931:Mitochondria, Muscle / Q000502:physiology*; D024101:Mitochondrial Proteins; D018482:Muscle, Skeletal / Q000166:cytology / Q000502:physiology*; D010957:Plasmids; D011506:Proteins / Q000235:genetics / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D013050:Spectrometry, Fluorescence; D000071246:Uncoupling Protein 2
D018076:DNA, Complementary; D007473:Ion Channels; D026901:Membrane Transport Proteins; D024101:Mitochondrial Proteins; D011506:Proteins; C000604391:Ucp2 protein, rat; D000071246:Uncoupling Protein 2