About the Redox Molecular Signaling Core

Redox Molecular Signaling Core

The Redox Molecular Signaling Core occupies 3,375 sq. ft. of laboratory space on the 3rd floor of the Medical School building and on the 6th floor of the adjacent Biomedical Research Institute. As part of the COBRE Center for Redox Biology and Cardiovascular Disease, this core is able to offer services at a significantly reduced rate to facilitate the study of redox regulation of cardiovascular disease at LSU Health – Shreveport. In addition, the COBRE grant provides money to expand this core facility over time, as discussed below. Currently, this core is divided functionally and spatially into two distinct sub-cores: the Analytic Redox Biology Sub-Core and the Molecular Signaling Sub-Core.

Analytical Redox Biology Sub-Core: The Analytic Redox Biology Sub-Core occupies space in Room F6-12 (690 sq. ft.) of the Biomedical Research Institute and Room 3-449 (1,074 sq. ft) in the adjacent Medical School Building. This core facility provides high quality, accurate measurements of reactive oxygen species, reactive nitrogen species, and reactive sulfide species in cell culture and tissue samples. High performance liquid chromatography (HPLC) systems coupled with UV-vis and fluorescence detectors are used to specifically quantify cellular and mitochondrial superoxide production, hydrogen sulfide pools (free sulfide, sulfide bound to transitional metals, and sulfane sulfur), and thiols (glutathione, GSH/GSSG, cysteine, cystine, homocysteine, persulfides, and glutathionylation). In addition, this core facility employs a versatile and highly sensitive Sievers NO Analyzer (NOA 280i, GE) capable of measuring nitric oxide, nitrite, nitrate, nitrosoheme, and nitrosothiols in a variety of biological samples. Lastly, this core utilizes a collection of mass spectrometery equipment, including a SYNAPT HDMS (Waters Corp.), a LCQ-DECA XP (Thermo), and a Q-TOF micro (Waters Corp.), to facilitate protein identification, identify post-translational modifications, and quantify fatty acid oxidation products.

Molecular Signaling Sub-Core: The Molecular Signaling Sub-Core is located on the 3rd floor of the Medical School Building in a bank of adjacent laboratories (3-236, 3-324, 3-326, 3-328, 3-330, 3-332, 3-334) occupying approximately 1,612 sq. ft. of laboratory space. The facility offers services for molecular cloning and site-directed mutagenesis, as well as design and production of vectors for CRISPR/Cas9 modification of vascular cells. Through the use of three dedicated cell culture rooms, this core provides services for endothelial, smooth muscle, and cardiac myocyte cell isolation, for generation of vascular cell lines, and for adenovirus and lentivirus production for transient or stable modification of cardiovascular cells. In addition, this core provides access to equipment and expertise for exposing vascular cells to hemodynamic forces using parallel plate flow chambers and access to equipment for the study of cellular effects of hypoxia/reoxygenation injury (Coy Hypoxic Chamber). As part of the ongoing COBRE grant, this core will purchase a hypoxia-capable CLARIOstar spectrofluorometer in 2020 to allow investigators to assess the cellular redox state over an extended, uninterrupted time course, including assessment of redox signaling during hypoxia and reoxygenation. In addition, this facility will purchase a ProteinSimple Wes Simple Western system in 2021 to provide investigators with an automated, high throughput analysis of changes in protein expression and function with reduced variability and improved quantitation compared to typical methods of manual Western blotting.

The Center for Redox Biology and Cardiovascular Disease is supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number P20GM121307.