A protocol for detecting the cytoplasm-exposed mitochondrial DNA

Mitochondria, being semi-autonomous organelles, possess a double membrane structure and harbor their own DNA, known as mtDNA. In situations of stress, mtDNA is released from the mitochondrial membrane and enters the cytoplasm. The mtDNA released into the cytoplasm plays a dual role in promoting both the initiation and escalation of intracellular reactive oxygen species (ROS), the activation of inflammatory pathways, and the death of cells. Consequently, the identification of intracytoplasmic mtDNA fragments holds immense significance in the realm of scientific investigation. In this paper, we delineate the experimental methodologies presently employed for quantifying intracytoplasmic mtDNA fragments. • Under various stress conditions, mitochondrial dysfunction can lead to the release of mitochondrial DNA (mtDNA) into the cytoplasm, triggering inflammatory pathways that contribute to the pathogenesis of diverse diseases. • This article provides an overview of the current experimental methodologies used to quantify intracytoplasmic mtDNA fragments and presents the data obtained from these experiments. • The methods outlined can serve as a valuable resource for researchers interested in studying abnormalities in mtDNA localization.