# Molecular and cellular mechanisms regulating mitochondrial subpopulation dynamics and function in vivo

> **NIH NIH R35** · OKLAHOMA MEDICAL RESEARCH FOUNDATION · 2024 · $437,000

## Abstract

Project Summary
Mitochondria regulate a number of critical cellular pathways including energy homeostasis, calcium handling
and lipid production. In a number of cell types, distinct populations of mitochondria are created and maintained
within subcellular compartments driving unique responses to physiological challenges in different regions of the
cell. While many of the molecular players that modulate mitochondrial shape, and therefore function, have
been identified, complete understanding of their functions and interactions in establishing these subpopulations
of mitochondria within cells remain difficult to define. The deficit in understanding subcellular mitochondrial
shape and function is largely due to a limited ability to visualize, and manipulate, these dynamic organelles in a
truly physiological environment at high spatial and temporal resolution. Our approaches are designed to
address these gaps in knowledge by leveraging newly developed technologies enabling genetic labelling and
manipulation, across multiple cell types, with high spatial and temporal imaging of mitochondrial morphology,
dynamics and function in vivo. In project one, members of the laboratory will target the four known mammalian
receptors (MFF, FIS1, MIEF1/2) of the dynamin-like protein one (DRP1), the main effector of mitochondrial
fission, to test their roles in the creation and maintenance of different mitochondrial subpopulations in cortical
neurons and skeletal myocytes in vivo. Through the use of loss of function experiments, CRISPR/Cas labeling
and targeting-motif analysis coupled with high resolution imaging we will map the molecular mechanisms
regulating subcellular mitochondrial fission dynamics across multiple mitochondrial subpopulations. In project
two, members of the laboratory will implement methods for sparse, bright labeling of cortical neuron and
skeletal myocyte mitochondria with fluorescent reporters for adenosine triphosphate, calcium, pH and reactive
oxygen species, and couple it with 2-photon imaging in living mice to reveal how these mitochondrial
subpopulations inform mitochondrial and cellular function in vivo. By manipulating different subpopulations and
visualizing the effects on mitochondrial and cellular function in multiple cell types in vivo, we will provide a
uniquely integrated approach to understanding the universal and cell-specific roles of mitochondrial
subpopulations found within cells.

## Key facts

- **NIH application ID:** 10833633
- **Project number:** 5R35GM137921-05
- **Recipient organization:** OKLAHOMA MEDICAL RESEARCH FOUNDATION
- **Principal Investigator:** Tommy L Lewis
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $437,000
- **Award type:** 5
- **Project period:** 2020-08-01 → 2025-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10833633

## Citation

> US National Institutes of Health, RePORTER application 10833633, Molecular and cellular mechanisms regulating mitochondrial subpopulation dynamics and function in vivo (5R35GM137921-05). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10833633. Licensed CC0.

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