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Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria

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 mutation s within the mitochondrial genome .

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Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria

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

  2. BackgroundMitochondrial DNA and Diseases • There is large number of diseases associated with mutationswithin the mitochondrial genome. • At the same time there is limited understanding of the mitochondiral-based disorders

  3. 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.

  4. Illustration of Heterogeneity [Heteroplasmy] of mtDNA http://hihg.med.miami.edu/code/http/modules/education/Design/CoursePageContent.asp?ID=20

  5. Method • Isolation individual mitochondrial from single cellsHL-60 • Performingmolecular analysis on that mitochondrion's genome.

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

  7. Results • 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   

  8. Comments

  9. 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.

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