Hybrid Gel Electrophoresis Using Skin Fibroblasts to Aid in Diagnosing Mitochondrial Disease

Overview

Journal Neurol Genet

Date 2019 Jun 14

PMID 31192304

Citations 2

Authors

Christopher Newell

Aneal Khan

David Sinasac

John Shoffner

Marisa W Friederich

Johan L K Van Hove

Stacey Hume

Jane Shearer

Iveta Sosova

Affiliations

Soon will be listed here.

Abstract

Objective: We developed a novel, hybrid method combining both blue-native (BN-PAGE) and clear-native (CN-PAGE) polyacrylamide gel electrophoresis, termed BCN-PAGE, to perform in-gel activity stains on the mitochondrial electron transport chain (ETC) complexes in skin fibroblasts.

Methods: Four patients aged 46-65 years were seen in the Metabolic Clinic at Alberta Children's Hospital and investigated for mitochondrial disease and had BN-PAGE or CN-PAGE on skeletal muscle that showed incomplete assembly of complex V (CV) in each patient. Long-range PCR performed on muscle-extracted DNA identified 4 unique mitochondrial DNA (mtDNA) deletions spanning the gene of CV. We developed a BCN-PAGE method in skin fibroblasts taken from the patients at the same time and compared the findings with those in skeletal muscle.

Results: In all 4 cases, BCN-PAGE in skin fibroblasts confirmed the abnormal CV activity found from muscle biopsy, suggesting that the mtDNA deletions involving were most likely germline mutations that are associated with a clinical phenotype of mitochondrial disease.

Conclusions: The BCN-PAGE method in skin fibroblasts has a potential to be a less-invasive tool compared with muscle biopsy to screen patients for abnormalities in CV and other mitochondrial ETC complexes.

Citing Articles

Recent advances in mitochondrial diseases: From molecular insights to therapeutic perspectives.

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Native Gel Electrophoresis and Immunoblotting to Analyze Electron Transport Chain Complexes.

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