TY - JOUR
T1 - Biochemical and molecular investigation of mitochondrial disease
T2 - an illustrative case showing the value of a multifaceted approach
AU - Byrne, E.
AU - Jean‐Francois, B.
AU - Thyagarajan, D.
AU - Collins, S.
AU - Dennett, X.
AU - Marzuki, S.
PY - 1991/12
Y1 - 1991/12
N2 - Abstract :Detailed biochemical and molecular investigations in a patient with Kearns‐Sayre syndrome are presented. Polarographic studies in isolated mitochondria revealed a global impairment in respiratory capacity consistent with an admixture of functional and non‐functional mitochondria. Cytochrome difference spectra revealed a selective deficiency in cytochrome aa3. Western immunoblot studies revealed normal subunit content of Complexes 1, III and IV. Southern blot studies of mtDNA showed a deletion of approximately 5 Kb coexisting with wild type DNA. PCR analysis confirmed that this deletion lies between the ATPase8 and NAD coenzyme Q oxidoreductase subunit 5 (ND5) genes. Breakpoint sequencing revealed a 13 nucleotide direct repeat flanking sequence (ACCTCCCTCACCA) consistent with slippage in mtDNA during replication as the mechanism of deletion. Histochemical studies of skeletal muscle revealed many cytochrome oxidase negative fibres and immunocytochemical studies showed cytochrome oxidase negative areas with abundant respiratory complex protein suggesting upregulation. The value of a multifaceted approach in unravelling the pathophysiology of mitochondrial diseases is emphasised. (Aust NZ J Med 1991; 21: 837–843.)
AB - Abstract :Detailed biochemical and molecular investigations in a patient with Kearns‐Sayre syndrome are presented. Polarographic studies in isolated mitochondria revealed a global impairment in respiratory capacity consistent with an admixture of functional and non‐functional mitochondria. Cytochrome difference spectra revealed a selective deficiency in cytochrome aa3. Western immunoblot studies revealed normal subunit content of Complexes 1, III and IV. Southern blot studies of mtDNA showed a deletion of approximately 5 Kb coexisting with wild type DNA. PCR analysis confirmed that this deletion lies between the ATPase8 and NAD coenzyme Q oxidoreductase subunit 5 (ND5) genes. Breakpoint sequencing revealed a 13 nucleotide direct repeat flanking sequence (ACCTCCCTCACCA) consistent with slippage in mtDNA during replication as the mechanism of deletion. Histochemical studies of skeletal muscle revealed many cytochrome oxidase negative fibres and immunocytochemical studies showed cytochrome oxidase negative areas with abundant respiratory complex protein suggesting upregulation. The value of a multifaceted approach in unravelling the pathophysiology of mitochondrial diseases is emphasised. (Aust NZ J Med 1991; 21: 837–843.)
KW - investigative techniques
KW - Mitochondrial diseases
KW - mitochondrial DNA abnormalities
UR - https://www.scopus.com/pages/publications/0026280502
U2 - 10.1111/j.1445-5994.1991.tb01404.x
DO - 10.1111/j.1445-5994.1991.tb01404.x
M3 - Article
C2 - 1818542
AN - SCOPUS:0026280502
SN - 0004-8291
VL - 21
SP - 837
EP - 843
JO - Australian and New Zealand Journal of Medicine
JF - Australian and New Zealand Journal of Medicine
IS - 6
ER -