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Evaluation of Gastrointestinal mtDNA Depletion in Mitochondrial Neurogastrointestinal Encephalomyopathy (MNGIE)

  • Carla GiordanoEmail author
  • Giulia d’Amati
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 755)

Abstract

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare disease characterized by severe gastro-intestinal (GI) dysmotility caused by mutations in the thymidine phosphorylase gene. Thymidine phosphorylase (TP) is involved in the control of the pyrimidine nucleoside pool of the cell. Reduced TP activity induces nucleotide pool imbalances that in turn affect both the rate and fidelity of mtDNA replication, leading to multiple deletions and depletion of mtDNA. By using laser capture microdissection and quantitative real-time-polymerase chain reaction technique, we showed that depletion of mitochondrial DNA (mtDNA) is the most prominent molecular defect in the gut wall of MNGIE patients. Depletion affects severely the smooth muscle cells of muscularis propria and the skeletal muscle component of the upper esophagus, while ganglion cells of the myenteric plexus show only a milder mtDNA reduction.

Key words

Mitochondrial disease mtDNA depletion mtDNA deletion CPEO 

Notes

Acknowledgments

The authors wish to thank Doctor Martina Leopizzi and Doctor Maurizia Orlandi for technical assistance. This work was supported by Fondazione Giuseppe Tomasello and Associazione Serena Tallarico per i Giovani nel Mondo.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Experimental MedicineSapienza UniversityRomeItaly

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