Published 1. 9. 2022
Following a short holiday break, we would like to invite you to the next institutional seminar of the IEM CAS, this time on the topic of Mitochondrial DNA damage and DNA repair capacity in type 2 diabetes obese rats.
Kristýna Tomášová, MSc. / Dept. of the Molecular Biology of Cancer
Metabolic syndrome represents a cluster of conditions, including insulin resistance, hypertension, obesity, and hyperglycemia. A characteristic of all these conditions is excess generating of reactive oxygen species, creating oxidative stress, and contributing to higher susceptibility to oxidative DNA damage, especially in mitochondria. As mitochondria play an important role in metabolism, efficient DNA repair to prevent the accumulation of DNA lesions is crucial for their proper functioning.
Our study aims to look closely at mitochondrial 8-oxoguanine DNA damage and repair in ZSF1 (Zucker fatty and spontaneously hypertensive) obese and -lean (control) rats, an experimental model suitable for studying type 2 diabetes. We intend to demonstrate if increased oxidative stress leads to mutagenic DNA lesions and compromised base excision repair capacity, specifically in mitochondria. Mitochondria were isolated from the liver using density gradient ultracentrifugation, and their protein purity was verified on Western blot. The protein extracts are now tested by alkaline Comet assay to analyze base excision repair activity in mitochondria. Mitochondrial DNA oxidative lesions will be detected using FPG (formamidopyrimidine [fapy]-DNA glycosylase) treatment and subsequent quantitative PCR.
The study was realized within the Mobility Program at the Department of Pharmacology and Toxicology, Maastricht University, the Netherlands, with which our Department has a long-term collaboration.