The goal of the Integrative Redox Biology Core is to provide investigators with a coordinated and comprehensive evaluation of the molecules, pathways, and systems that contribute to oxidative metabolism and oxidative stress using analyses of frozen samples from a wide variety of model species (e.g. invertebrates, rodents, exceptionally long-lived species, and humans). These analyses include state-of-the-art measures of redox and energy balance as well as oxidative damage and mitochondrial function. The Core will offer accurate and sensitive analyses of altered redox status and ATP levels as well as mitochondrial function and oxidative damage through a coordinated evaluation of a number of molecules, pathways, and systems. Redox status and oxidant homeostasis have a major impact on physiologic processes underlying aging and contribute to a number of age-related diseases, including diabetes, cancer and neurodegenerative disease. Thus, these assays will be valuable for a wide range of investigators studying aging and age-related disease.
The Specific Aims of the Integrative Redox Biology Core are:
Aim 1. To perform accurate and comprehensive analyses of redox and energetic status. The core will provide analyses of the following redox couples: GSH/GSSG, NADPH/NADP+, NADH/NAD+, thioredoxin (-SH/S-S), peroxiredoxin (-SH/S-S), CoASH/Acetyl-CoA, as well as the energy charge (ATP, ADP, & AMP).
Aim 2. To determine consequences of changes in oxidative stress by measuring oxidative modifications and damage to cellular components as well as the expression profiles of antioxidant proteins. Oxidative damage to cell components will be analyzed using assays we have developed; oxidative damage to lipid, protein, and DNA will be measured by F2- isoprostanes, protein carbonyls, and 8-oxo-dG, respectively.
Aim 3. To provide additional assays of mitochondrial function using permeabilized tissue, isolated mitochondria, and cell culture models. The Core will provide services to measure mitochondrial function e.g, ETC activities, oxidant generation, ATP production, and respiration in fresh tissue or isolated mitochondria. A significant strength of the Core is that, because we are analyzing levels of highly specific biological compounds, our assays can be used with tissues or cells from any living organism. In addition, the analyses we provide are especially appropriate for a Core because they require costly equipment and significant technologic expertise that can limit these types of analyses as routine measures in individual laboratories. Overall, the key advantage of this core is that we can provide a comprehensive, accurate, and sensitive set of redox and oxidative stress related measurements in a wide variety of organisms with frozen tissue that can be shipped from any laboratory in the United States.
The Integrative Redox Biology Core will provide investigators with a coordinated and comprehensive analysis of the molecules, pathways, and systems that contribute to oxidative metabolism and oxidative stress using frozen samples from a wide variety of model species and state-of-the-art assays. Assays of redox couples, energy status, oxidative damage and mitochondrial function will allow investigators to probe pathways related to aging and age- related disease.
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