A nanoscale, multi-parametric flow cytometry-based platform to study mitochondrial heterogeneity and mitochondrial DNA dynamics
| Autor: | Alisha M. Bothun, Alexander R. Ivanov, Julie A. MacDonald, Konstantin Khrapko, Dori C. Woods, Jonathan L. Tilly, Somak Ray, Yuanwei Gao, Sofia Annis, Hannah Sheehan |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Male
Proteomics Programmed cell death Mitochondrial DNA Cell biology Cell Medicine (miscellaneous) Cell Separation Computational biology Mitochondrion Biology DNA Mitochondrial Mitochondrial Dynamics General Biochemistry Genetics and Molecular Biology Article Flow cytometry Mice 03 medical and health sciences Adenosine Triphosphate 0302 clinical medicine Organelle medicine Animals Nanotechnology lcsh:QH301-705.5 Fluorescent Dyes 030304 developmental biology 0303 health sciences medicine.diagnostic_test Proteomic Profiling Biological techniques Brain Flow Cytometry Mitochondria Mice Inbred C57BL Microscopy Electron medicine.anatomical_structure lcsh:Biology (General) Calibration Female General Agricultural and Biological Sciences 030217 neurology & neurosurgery Function (biology) |
| Zdroj: | Communications Biology Communications Biology, Vol 2, Iss 1, Pp 1-13 (2019) |
| ISSN: | 2399-3642 |
| Popis: | Mitochondria are well-characterized regarding their function in both energy production and regulation of cell death; however, the heterogeneity that exists within mitochondrial populations is poorly understood. Typically analyzed as pooled samples comprised of millions of individual mitochondria, there is little information regarding potentially different functionality across subpopulations of mitochondria. Herein we present a new methodology to analyze mitochondria as individual components of a complex and heterogeneous network, using a nanoscale and multi–parametric flow cytometry-based platform. We validate the platform using multiple downstream assays, including electron microscopy, ATP generation, quantitative mass-spectrometry proteomic profiling, and mtDNA analysis at the level of single organelles. These strategies allow robust analysis and isolation of mitochondrial subpopulations to more broadly elucidate the underlying complexities of mitochondria as these organelles function collectively within a cell. Julie MacDonald et al. present a method for mitochondrial isolation using fluorescence-activated mitochondria sorting to facilitate analysis from individual organelles. They validate their platform using a number of downstream assays, including electron microscopy, proteomic profiling, and mtDNA analysis. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|