title: Microanalysis of cardiolipin in small biopsies including skeletal muscle from patients with mitochondrial disease.
abstract: Cardiolipin is a specific mitochondrial phospholipid that is present in mammalian tissues in low concentration. To measure cardiolipin in small biopsies from patients with mitochondrial disease, we developed a new technique that can detect subnanomolar levels of well-resolved molecular species, the most abundant of which are tetralinoleoyl-cardiolipin (L(4)) and trilinoleoyl-oleoyl-cardiolipin (L(3)O). To this end, a fluorescence-labeled derivative of cardiolipin (2-[naphthyl-1'-acetyl]-cardiolipin dimethyl ester) was formed and analyzed by high performance liquid chromatography. Cardiolipin was measured in skeletal muscle biopsies from 8 patients with mitochondrial disease and in 17 control subjects. In 5 patients with mitochondrial disease, cardiolipin content was higher than normal (2. 4;-7.0 vs. 0.4;-2.2 nmol/mg protein). In 3 patients with mitochondrial disease, the L(4)/L(3)O ratio was lower than normal (2;-4 vs. 4;-6). Cardiolipin was also measured in various rat and dog muscle tissues. The L(4)/L(3)O ratio was higher in condensed "muscle" type mitochondria (heart ventricle, skeletal muscle, ratios 4;-7) than in orthodox "liver" type mitochondria (liver, smooth muscle, heart auricular appendage, H9c2 myoblasts, ratios 0.4;-3), suggesting that the L(4)/L(3)O proportion is important for cristae membrane structure. We concluded that the L(4)/L(3)O ratio is a tissue-specific variable that may change in the presence of mitochondrial disease. The new method is suitable to measure cardiolipin in muscle biopsies in order to estimate concentration of mitochondria.
authors.affiliation: Department of Anesthesiology, Hospital for Special Surgery, Cornell University Medical College, 535 E. 70th Street, New York, NY.
authors.forename: M
authors.initials: M
authors.lastname: Schlame
authors.forename: S
authors.initials: S
authors.lastname: Shanske
authors.forename: S
authors.initials: S
authors.lastname: Doty
authors.forename: T
authors.initials: T
authors.lastname: König
authors.forename: T
authors.initials: T
authors.lastname: Sculco
authors.forename: S
authors.initials: S
authors.lastname: DiMauro
authors.forename: T J
authors.initials: TJ
authors.lastname: Blanck
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D001706:Biopsy; D002308:Cardiolipins / Q000032:analysis* / Q000737:chemistry; D002648:Child; D002675:Child, Preschool; D004285:Dogs; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D007223:Infant; D008297:Male; D008832:Microchemistry / Q000379:methods*; D008875:Middle Aged; D008929:Mitochondria, Heart / Q000648:ultrastructure; D008931:Mitochondria, Muscle / Q000648:ultrastructure; D017240:Mitochondrial Myopathies / Q000378:metabolism* / Q000473:pathology; D008958:Models, Molecular; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002308:Cardiolipins; D005456:Fluorescent Dyes