Mutations in human mitochondrial DNA (mtDNA) can cause mitochondrial disease and have been associated with neurodegenerative disorders, cancer, diabetes and aging. Yet our progress toward delineating the precise contributions of mtDNA mutations to these conditions is impeded by the limited availability of faithful transmitochondrial animal models. Here, we report a method for the isolation of mutations in mouse mtDNA and its implementation for the generation of a collection of over 150 cell lines suitable for the production of transmitochondrial mice. This method is based on the limited mutagenesis of mtDNA by proofreading-deficient DNA-polymerase γ followed by segregation of the resulting highly heteroplasmic mtDNA population by means of intracellular cloning. Among generated cell lines, we identify nine which carry mutations affecting the same amino acid or nucleotide positions as in human disease, including a mutation in the ND4 gene responsible for 70% of Leber Hereditary Optic Neuropathies (LHON). Similar to their human counterparts, cybrids carrying the homoplasmic mouse LHON mutation demonstrated reduced respiration, reduced ATP content and elevated production of mitochondrial reactive oxygen species (ROS). The generated resource of mouse mtDNA mutants will be useful both in modeling human mitochondrial disease and in understanding the mechanisms of ROS production mediated by mutations in mtDNA.
Sediment remains one of the most commonly occurring pollutants affecting the U.S.’s water bodies, as identified by the United States Environmental Protection Agency (USEPA) ( 1). Construction activities largely accelerate soil erosion and subsequent sediment deposition. The National Pollutant Discharge Elimination System Construction General Permit requires construction operators to implement erosion and sediment control (E&SC) plans to minimize downstream implications from sediment-laden discharge. However, E&SC practices are often designed from “rules of thumb” and lack scientific, performance-based evidence in their design and implementation. The Auburn University Stormwater Research Facility (AU-SRF), previously the Auburn University Erosion and Sediment Control Testing Facility (AU-ESCTF), is an outdoor research center dedicated to evaluating E&SC practices and products commonly used on highway construction projects. Large-scale test apparatuses and methods at AU-SRF are designed to mimic construction site conditions, including rainfall, flow rates, topography, and soil characteristics, to evaluate existing and novel E&SC practices. Since its inception in 2008, AU-SRF has provided small-, medium-, and large-scale testing evaluations for numerous Departments of Transportation and product manufacturers. Findings from controlled testing have continued to inform the selection, design, implementation, and maintenance of E&SC practices used on construction sites and protect downstream waters and infrastructure. In the first decade, AU-SRF has directed 13 research projects and produced more than 30 peer-reviewed publications and 100 professional presentations. As AU-SRF grows into its second decade and efforts reach outside of the southeastern region, the mission to advance knowledge through E&SC research and development, product evaluation, and training remains constant. This review synthesizes the research produced from large-scale testing at AU-SRF to date and presents ongoing projects.
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