# High throughput assay for mitochondrial respiration in aged brain microvessels

> **NIH NIH R21** · TULANE UNIVERSITY OF LOUISIANA · 2020 · $202,875

## Abstract

Aging is a major contributor to cerebrovascular disease and subsequent neurological sequelae. The
frequency and severity of these diseases, age of onset, and underlying mechanisms differ between men and
women and are augmented by age-related diseases such as type 2 diabetes (T2D). The endothelium, a
principle component of the microcirculation, is particularly sensitive to the negative effects of aging or T2D and
mitochondria appear to play a pivotal role. Progress in elucidating the underlying mechanisms negatively
affecting endothelial mitochondria and developing beneficial therapies has been obstructed due to the inability
to follow mitochondrial characteristics in the brain microcirculation in real time during the development of aging
and age-associated diseases. To address this deficiency, we will use three new approaches. First, we will use
a mouse model which we developed with genetic labeling of mitochondria only in endothelium with Dendra2
fluorescent protein (mitoDendr2 FP). Second, we will use a novel, high throughput method we developed that
allows the determination of energy production by mitochondrial respiration and glycolysis in freshly harvested
brain microvessel from the mouse. Glycolysis is a major energy producing process in the endothelium and the
relative importance of oxidative phosphorylation (OXPHOS) and glycolysis changes with aging and T2D is
unclear. We also will examine whether mitochondrial fuels change. Third, we will use RNA Sequencing and
Proteomics to explore underlying mechanisms involving energy producing pathways. Our preliminary and
published data have led to the overall hypothesis that mitochondria play key roles in adverse changes
in the cerebral microcirculation during aging and T2D and that therapies targeting mitochondria are
protective. Thus, we speculate that mitochondria in microvessels are adversely affected more and earlier than
large arteries during aging, T2D acerbates changes in mitochondria in microvessels, glycolysis becomes a
more important source of ATP during aging and T2D, alternative fuel sources for OXPHOS become important
during aging and T2D, mitochondria are more resilient in female microvessels, and mitochondria represent a
useful therapeutic target to protect microvessels. We have 2 aims. Aim 1: Elucidate mechanisms of
mitochondrial and vascular changes during aging. We will: a) determine mitochondrial and vascular
characteristics in vivo in male and female aged mice, b) determine effects of aging on glycolysis and
OXPHOS, mitochondrial fuel, and mitoROS, c) elucidate mechanisms affecting mitochondrial and glycolytic
dynamics, and d) explore treatment modalities. Aim 2: Determine mechanisms of mitochondrial and
vasculature changes during aging and T2D. We will: a) determine mitochondrial and vascular
characteristics in aged male and female mice during T2D, b) determine effects of aging on glycolysis and
OXPHOS, mitochondrial fuel, and mitoROS, c) elucidate mechanisms involved in chang...

## Key facts

- **NIH application ID:** 9980261
- **Project number:** 5R21AG063345-02
- **Recipient organization:** TULANE UNIVERSITY OF LOUISIANA
- **Principal Investigator:** DAVID W BUSIJA
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $202,875
- **Award type:** 5
- **Project period:** 2019-08-01 → 2021-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9980261

## Citation

> US National Institutes of Health, RePORTER application 9980261, High throughput assay for mitochondrial respiration in aged brain microvessels (5R21AG063345-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9980261. Licensed CC0.

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