Analysis of Mitochondrial Network Morphology in Cultured Myoblasts from Patients with Mitochondrial Disorders

Abstract

Mitochondrial morphology was studied in cultivated myoblasts obtained from patients with mitochondrial disorders, including CPEO, MELAS and TMEM70 deficiency. Mitochondrial networks and ultrastructure were visualized by fluorescence microscopy and transmission electron microscopy, respectively. A heterogeneous picture of abnormally sized and shaped mitochondria with fragmentation, shortening, and aberrant cristae, lower density of mitochondria and an increased number of “megamitochondria” were found in patient myoblasts. Morphometric Fiji analyses revealed different mitochondrial network properties in myoblasts from patients and controls. The small number of cultivated myoblasts required for semiautomatic morphometric image analysis makes this tool useful for estimating mitochondrial disturbances in patients with mitochondrial disorders.

• Cristae
• Fiji
• image analysis
• mitochondrial disorders
• myoblasts
• ultrastructure

Acknowledgements

The authors are very grateful to Dr. László Wenchich, Department of Pediatrics and Adolescent Medicine, and Eva Kirchmannová and Ivana Odvodyová, Institute of Pathology, First Faculty of Medicine, Charles University, Prague, Czech Republic, for their excellent technical assistance and to Dr. Jiří Janáček, Institute of Physiology CAS, v.v.i., Videnska 1083, Prague 4 - Krč, 14220, for his comments and ideas regarding the applied image analysis approaches and methods.

Funding

The study was supported by the Grant Agency of the Czech Republic (14-36804G), Charles University in Prague (PRVOUK P24/LF1/3, SVVUK 260148/2015 and UNCE 204011), the Ministry of Health of the Czech Republic (RVO-VFN 64165/2012 and RVO-67985823), the American Science and Information Center (LH13028).

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.