Severe depletion of mitochondrial DNA in spinal muscular atrophy.
Spinal muscular atrophy (SMA) is a neuromuscular disorder in childhood leading to a dramatic loss of muscle strength. Functional investigations with high-resolution polarography and enzyme measurements of the respiratory chain revealed lowered activities in muscle tissue of SMA patients. To gain a better understanding of this low energy supply we analyzed the amount of mitochondrial DNA (mtDNA) in skeletal muscle of 20 unrelated children with genetically proven SMA and 31 controls. Quantitative Southern blot analysis revealed a severe and homogeneous decrease in the content of muscle mtDNA in relation to nuclear DNA in SMA patients (90.3+/-7.8%), whereas by immunofluorescence no decrease in the number of mitochondria was detected. In addition, a two- to threefold reduction of the nuclear-encoded complex II (succinate dehydrogenase) activity was detected in SMA muscle tissue. Western blot analysis showed a significant reduction of both mitochondrial- and nuclear-encoded cytochrome c oxidase subunits. Our results indicate that mtDNA depletion in SMA is a consequence of severe atrophy, and has to be differentiated by measurement of complex II from an isolated reduction of mtDNA as found in patients with mitochondriocytopathies and the so-called mtDNA depletion syndrome.
Berger
Alexandra
A
Department of Pediatrics, General Hospital Salzburg, Muellner Hauptstrasse 48, Austria.
Mayr
Johannes A
JA
Meierhofer
David
D
Fötschl
Ulrike
U
Bittner
Reginald
R
Budka
Herbert
H
Grethen
Claude
C
Huemer
Michael
M
Kofler
Barbara
B
Sperl
Wolfgang
W
D000293:Adolescent; D000328:Adult; D000368:Aged; D015139:Blotting, Southern; D015153:Blotting, Western; D002648:Child; D002675:Child, Preschool; D004272:DNA, Mitochondrial / Q000032:analysis*; D003576:Electron Transport Complex IV / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007223:Infant; D008875:Middle Aged; D008928:Mitochondria / Q000201:enzymology / Q000473:pathology*; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000201:enzymology; D009134:Muscular Atrophy, Spinal / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D013385:Succinate Dehydrogenase / Q000378:metabolism
D004272:DNA, Mitochondrial; D013385:Succinate Dehydrogenase; D003576:Electron Transport Complex IV