Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria Summary and Discussion Mustafa Albahrani

Background Mitochondrial DNA and Diseases There is large number of diseases associated with mutations within the mitochondrial genome. At the same time there is limited understanding of the mitochondiral-based disorders

The Research Paper Purpose Understand further about the development of diseases related to mitochondria by applying a new method that measures the heterogeneity of mtDNA. Side Note—Heterogeneity [Heteroplasmy] of mtDNA The situation in which, within a single cell, there is a mixture of mitochondria (energy producing cytoplasmic organelles), some containing mutant DNA and some containing normal DNA The researcher stated that they want to further understand the the development of diseases related to mitochondria. Clai No solid understanding that could explain what effect the distribution of mutated mtDNA through the mitochondrial matrix has on mitochondrial diseases.

Illustration of Heterogeneity [Heteroplasmy] of mtDNA on/Design/CoursePageContent.asp?ID=20

Method - Isolation individual mitochondrial from single cells HL-60 - Performing molecular analysis on that mitochondrion's genome.

Results 1. Extraction of a single mitochondrion particle - Staining the mitochondrial membranes with indicator dye [Mitotracker green] - Lysing [breaking down] the cell using UV laser, and optical tweezers to trap an organelle. Stained mitochondira in Green

Results 2. Amplification and sequencing mtDNA from a single mitochondrion - 20 mitochondrial particles were captured and sequenced its genome. - the researcher found that the sequencing chromatograms of five mitDNA samples show heteroplasmy at np=12071

Comments

References Reiner, J. E., Kishore, R. B., Levin, B. C., Albanetti, T., Boire, N., Knipe, A.,... & Deckman, K. H. (2010). Detection of heteroplasmic mitochondrial DNA in single mitochondria. PloS one, 5(12), e14359.