Multicentric Standardization of Protocols for the Diagnosis of Human Mitochondrial Respiratory Chain Defects
Nuria Bujan,
Constanza Morén,
Francesc J. García-García,
Alberto Blázquez,
Clara Carnicer,
Ana Belén Cortés,
Cristina González,
Ester López-Gallardo,
Ester Lozano,
Sonia Moliner,
Laura Gort,
Ester Tobías,
Aitor Delmiro,
Miguel Ángel Martin,
Miguel Ángel Fernández-Moreno,
Eduardo Ruiz-Pesini,
Elena Garcia-Arumí,
Juan Carlos Rodríguez-Aguilera,
Glòria Garrabou
Affiliations
Nuria Bujan
Inborn Errors of Metabolism Section (IBC U737), Molecular Genetics and Biochemistry Service, Hospital Clínic of Barcelona, IDIBAPS, 08036 Barcelona, Spain
Constanza Morén
Muscle Research and Mitochondrial Function Laboratory (U722), Cellex-IDIBAPS, Faculty of Medicine and Health Sciences-University of Barcelona, Internal Medicine Department-Hospital Clínic of Barcelona, 08036 Barcelona, Spain
Francesc J. García-García
Muscle Research and Mitochondrial Function Laboratory (U722), Cellex-IDIBAPS, Faculty of Medicine and Health Sciences-University of Barcelona, Internal Medicine Department-Hospital Clínic of Barcelona, 08036 Barcelona, Spain
Alberto Blázquez
Mitochondrial Diseases Laboratory (U723), Research Institute, Universitary Hospital 12 de Octubre (Imas12), 28041 Madrid, Spain
Clara Carnicer
Laboratory of Inborn Errors of Metabolism, Biochemistry Service (U701), Laboratoris Clínics, Universitary Hospital Vall d’Hebron, 08035 Barcelona, Spain
Ana Belén Cortés
Bioenergetics and Cell Physiology Service (U729), Central Services of Research, University Pablo de Olavide, 41013 Sevilla, Spain
Cristina González
Biochemistry Department (U717), Biomedical Research Institute ‘Alberto Sols’, CSIC, Faculty of Medicine, Autonomous University of Madrid, and Instituto de Investigación Sanitaria Hospital 12 de Octubre (Imas12), 28041 Madrid, Spain
Ester López-Gallardo
Cell and Molecular Biology Department (U727), Veterinary Faculty, University of Zaragoza, 50009 Zaragoza, Spain
Ester Lozano
Department of Cell Biology, Physiology and Immunology, Faculty of Biology, Universitat de Barcelona (UB), and Institute of Biomedicine of the University of Barcelona (IBUB), 08007 Barcelona, Spain
Sonia Moliner
Inborn Errors of Metabolism Section (IBC U737), Molecular Genetics and Biochemistry Service, Hospital Clínic of Barcelona, IDIBAPS, 08036 Barcelona, Spain
Laura Gort
Inborn Errors of Metabolism Section (IBC U737), Molecular Genetics and Biochemistry Service, Hospital Clínic of Barcelona, IDIBAPS, 08036 Barcelona, Spain
Ester Tobías
Muscle Research and Mitochondrial Function Laboratory (U722), Cellex-IDIBAPS, Faculty of Medicine and Health Sciences-University of Barcelona, Internal Medicine Department-Hospital Clínic of Barcelona, 08036 Barcelona, Spain
Aitor Delmiro
Mitochondrial Diseases Laboratory (U723), Research Institute, Universitary Hospital 12 de Octubre (Imas12), 28041 Madrid, Spain
Miguel Ángel Martin
Mitochondrial Diseases Laboratory (U723), Research Institute, Universitary Hospital 12 de Octubre (Imas12), 28041 Madrid, Spain
Miguel Ángel Fernández-Moreno
Biochemistry Department (U717), Biomedical Research Institute ‘Alberto Sols’, CSIC, Faculty of Medicine, Autonomous University of Madrid, and Instituto de Investigación Sanitaria Hospital 12 de Octubre (Imas12), 28041 Madrid, Spain
Eduardo Ruiz-Pesini
Cell and Molecular Biology Department (U727), Veterinary Faculty, University of Zaragoza, 50009 Zaragoza, Spain
Elena Garcia-Arumí
Mitochondrial Pathology Laboratory, Research Institute, Universitary Hospital Vall d’Hebron, 08035 Barcelona, Spain
Juan Carlos Rodríguez-Aguilera
Bioenergetics and Cell Physiology Service (U729), Central Services of Research, University Pablo de Olavide, 41013 Sevilla, Spain
Glòria Garrabou
Muscle Research and Mitochondrial Function Laboratory (U722), Cellex-IDIBAPS, Faculty of Medicine and Health Sciences-University of Barcelona, Internal Medicine Department-Hospital Clínic of Barcelona, 08036 Barcelona, Spain
The quantification of mitochondrial respiratory chain (MRC) enzymatic activities is essential for diagnosis of a wide range of mitochondrial diseases, ranging from inherited defects to secondary dysfunctions. MRC lesion is frequently linked to extended cell damage through the generation of proton leak or oxidative stress, threatening organ viability and patient health. However, the intrinsic challenge of a methodological setup and the high variability in measuring MRC enzymatic activities represents a major obstacle for comparative analysis amongst institutions. To improve experimental and statistical robustness, seven Spanish centers with extensive experience in mitochondrial research and diagnosis joined to standardize common protocols for spectrophotometric MRC enzymatic measurements using minimum amounts of sample. Herein, we present the detailed protocols, reference ranges, tips and troubleshooting methods for experimental and analytical setups in different sample preparations and tissues that will allow an international standardization of common protocols for the diagnosis of MRC defects. Methodological standardization is a crucial step to obtain comparable reference ranges and international standards for laboratory assays to set the path for further diagnosis and research in the field of mitochondrial diseases.