There is significant evidence linking mitochondria to biological aging, with a recent theory proposing that giant mitochondria are involved in the aging of post-mitotic tissues. In these tissues, giant mitochondria appear to evade autophagocytosis, accumulate, and replace normal mitochondria. Techniques to characterize giant mitochondria in the presence of, or after isolation from, normal mitochondria are greatly needed. The short- term goal of this proposal is to characterize and isolate giant mitochondria and establish whether or not these mitochondria affect muscle function. The long-term goal is to use and make available novel technological approaches for uncovering the changes that, at the subcellular and molecular level, define biological human aging. The proposed work utilizes mitochondria from a myoblast model and muscle from Fischer 344 rats. We have four aims: (1) Characterize giant mitochondria in vivo in cell models based on their cell biology. (2) Purify giant mitochondria isolated from these in vivo cell models and then characterize these giant mitochondria biochemically. (3) Compare the proteomes of giant and normal mitochondria isolated from these models. (4) Characterize giant mitochondria in aging skeletal muscle.
The first aim uses confocal microscopy, or this technique combined with single cell capillary electrophoresis, to investigate size, DMAcontent and synthesis, DNA mutations, carbonyl accumulation, ROS production, membrane potential, mitochondrial fusion, and autophagocytosis.
The second aim describes the methods to isolate subpopulations of giant and normal mitochondria using free-flow electrophoresis and then characterize these isolated mitochondria using methods suitable for isolated organelles.
The third aim utilizes stable isotope labeling in cell cultures to compare the proteomes of giant and normal mitochondria. The last aim investigates age-related changes in muscle containing giant mitochondria. Significance to Health. Giant mitochondria remain poorly characterized, their proteomic and genomic characteristics have not been defined, and their relationship to muscle function has not been investigated. The characterization of this class of mitochondria is highly relevant to the public health not only because of their possible role in aging tissues, but also because of their appearance as a consequence of alcoholism, human toxicity, and multiple diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
3R01AG020866-05A1S1
Application #
7432916
Study Section
Enabling Bioanalytical and Biophysical Technologies Study Section (EBT)
Program Officer
Finkelstein, David B
Project Start
2002-04-15
Project End
2012-03-31
Budget Start
2007-06-15
Budget End
2008-03-31
Support Year
5
Fiscal Year
2007
Total Cost
$23,202
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Muratore, Katherine A; Najt, Charles P; Livezey, Nicholas M et al. (2018) Sizing lipid droplets from adult and geriatric mouse liver tissue via nanoparticle tracking analysis. Anal Bioanal Chem 410:3629-3638
Satori, Chad P; Ramezani, Marzieh; Koopmeiners, Joseph S et al. (2017) Checkpoints for Preliminary Identification of Small Molecules found Enriched in Autophagosomes and Activated Mast Cell Secretions Analyzed by Comparative UPLC/MSe. Anal Methods 9:46-54
Daniele, Joseph R; Esping, Daniel J; Garcia, Gilbert et al. (2017) ""High-Throughput Characterization of Region-Specific Mitochondrial Function and Morphology"". Sci Rep 7:6749
Muratore, Katherine A; Grundhofer, Heather M; Arriaga, Edgar A (2016) Capillary Electrophoresis with Laser-Induced Fluorescent Detection of Immunolabeled Individual Autophagy Organelles Isolated from Liver Tissue. Anal Chem 88:11691-11698
Luo, Jinghui; Muratore, Katherine A; Arriaga, Edgar A et al. (2016) Deterministic Absolute Negative Mobility for Micro- and Submicrometer Particles Induced in a Microfluidic Device. Anal Chem 88:5920-7
Palsuledesai, Charuta C; Ochocki, Joshua D; Kuhns, Michelle M et al. (2016) Metabolic Labeling with an Alkyne-modified Isoprenoid Analog Facilitates Imaging and Quantification of the Prenylome in Cells. ACS Chem Biol 11:2820-2828
Daniele, Joseph R; Heydari, Kartoosh; Arriaga, Edgar A et al. (2016) Identification and Characterization of Mitochondrial Subtypes in Caenorhabditis elegans via Analysis of Individual Mitochondria by Flow Cytometry. Anal Chem 88:6309-16
Kumar, Shailabh; Wolken, Gregory G; Wittenberg, Nathan J et al. (2015) Nanohole Array-Directed Trapping of Mammalian Mitochondria Enabling Single Organelle Analysis. Anal Chem 87:11973-7
Luo, Jinghui; Abdallah, Bahige G; Wolken, Gregory G et al. (2014) Insulator-based dielectrophoresis of mitochondria. Biomicrofluidics 8:021801
Hahn, Wendy S; Kuzmicic, Jovan; Burrill, Joel S et al. (2014) Proinflammatory cytokines differentially regulate adipocyte mitochondrial metabolism, oxidative stress, and dynamics. Am J Physiol Endocrinol Metab 306:E1033-45

Showing the most recent 10 out of 58 publications