MITOCHONDRIAL DISORDERS: BIOCHEMICAL AND MOLECULAR ANALYSIS

By EMMA HARRIS

CONTENTS PART I MITOCHONDRIAL DISORDER: A COMPLEX DISEASE OF THE TWO GENOMES 1 MITOCHONDRIAL DNA MUTATIONS: AN OVERVIEW OF CLINICAL AND MOLECULAR ASPECTS 2 NUCLEAR GENE DEFECTS IN MITOCHONDRIAL DISORDERS 3 DIAGNOSTIC CHALLENGES OF MITOCHONDRIAL DISORDERS: COMPLEXITIES OF TWO GENOMES PART II BIOCHEMICAL ANALYSIS OF MITOCHONDRIAL DISORDERS 4 BIOCHEMICAL ANALYSES OF THE ELECTRON TRANSPORT CHAIN COMPLEXES BY SPECTROPHOTOMETRY 5 MEASUREMENT OF MITOCHONDRIAL OXYGEN CONSUMPTION USING A CLARK ELECTRODE 6 MITOCHONDRIAL RESPIRATORY CHAIN: BIOCHEMICAL ANALYSIS AND CRITERION FOR DEFICIENCY IN DIAGNOSIS 7 ASSAYS OF PYRUVATE DEHYDROGENASE COMPLEX AND PYRUVATE CARBOXYLASE ACTIVITY 8 ASSESSMENT OF THYMIDINE PHOSPHORYLASE FUNCTION: MEASUREMENT OF PLASMA THYMIDINE (AND DEOXYURIDINE) AND THYMIDINE PHOSPHORYLASE ACTIVITY 9 MEASUREMENT OF MITOCHONDRIAL DNTP POOLS 10 MEASUREMENT OF OXIDIZED AND REDUCED COENZYME Q IN BIOLOGICAL FLUIDS, CELLS, AND TISSUES: AN HPLC-EC METHOD 11 ASSAY TO MEASURE OXIDIZED AND REDUCED FORMS OF COQ BY LC–MS/MS SI HOUN HAHN, SANDRA KERFOOT, AND VALERIA VASTA 12 MORPHOLOGICAL ASSESSMENT OF MITOCHONDRIAL RESPIRATORY CHAIN FUNCTION ON TISSUE SECTIONS 13 BLUE NATIVE POLYACRYLAMIDE GEL ELECTROPHORESIS: A POWERFUL DIAGNOSTIC TOOL FOR THE DETECTION OF ASSEMBLY DEFECTS IN THE ENZYME COMPLEXES OF OXIDATIVE PHOSPHORYLATION 14 RADIOACTIVE LABELING OF MITOCHONDRIAL TRANSLATION PRODUCTS IN CULTURED CELLS 15 TRANSMITOCHONDRIAL CYBRIDS: TOOLS FOR FUNCTIONAL STUDIES OF MUTANT MITOCHONDRIA 16 FLUORESCENCE-ACTIVATED CELL SORTING ANALYSIS OF MITOCHONDRIAL CONTENT, MEMBRANE POTENTIAL, AND MATRIX OXIDANT BURDEN IN HUMAN LYMPHOBLASTOID CELL LINES 17 MOLECULAR PROFILING OF MITOCHONDRIAL DYSFUNCTION IN CAENORHABDITIS ELEGANS PART III MOLECULAR ANALYSIS OF MITOCHONDRIAL DISORDERS 18 ANALYSIS OF COMMON MITOCHONDRIAL DNA MUTATIONS BY ALLELE-SPECIFIC OLIGONUCLEOTIDE AND SOUTHERN BLOT HYBRIDIZATION 19 SEQUENCE ANALYSIS OF THE WHOLE MITOCHONDRIAL GENOME AND NUCLEAR GENES CAUSING MITOCHONDRIAL DISORDERS 20 UTILITY OF ARRAY CGH IN MOLECULAR DIAGNOSIS OF MITOCHONDRIAL DISORDERS 21 QUANTIFICATION OF MTDNA MUTATION HETEROPLASMY (ARMS QPCR) 22 MEASUREMENT OF MITOCHONDRIAL DNA COPY NUMBER 23 DETERMINATION OF THE CLINICAL SIGNIFICANCE OF AN UNCLASSIFIED VARIANT