Kvantitativní aspekty mitochondriálního genomu v diabetu a rakovině

Alán, Lukáš

Mitochondria are cellular powerhouses and physiological regulators with many features resembling their prokaryotic ancestors. They maintain their own mitochondrial DNA (mtDNA) encoding 13 inner mitochondrial membrane proteins of oxidative phosphorylation machinery, 22 transfer RNAs and two ribosomal RNAs. Healthy mitochondria in normal cells form a dynamic network consisting of highly interconnected tubules. The mitochondrial disorders are very heterogeneous and difficult to diagnose and cure. It is due to combining products of two genomes, the complexity of mitochondrial structure and due to insufficiency of current state of knowledge. To understand the mitochondrial nucleic acid species distribution, we have developed and established new techniques to visualize mitochondrial network, nucleoids and different RNA species together with qPCR techniques for monitoring the mitochondrial intactness. We determined nucleoid distribution and mtDNA amount following rotenone mediated respiratory inhibition or following degeneration of the electrical component of the protonmotive force by valinomycin treatment. Native mitochondria were mostly tubular with average nucleoid spacing 1.1 +/- 0.2µm which we termed as a nucleoid code. Subsequently induced fission resulted in mitochondrial network fragmentation and each...

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