title: Analysis of targeted mutation in DJ-1 on cellular function in primary astrocytes.
abstract: DJ-1 mutation induces early-onset Parkinson's disease, and conversely over-expression of DJ-1 is associated with cancer in numerous tissues. A gene-trap screening library conducted in embryonic stem cells was utilized for generation of a DJ-1 mutant mouse. Real-time PCR and immunoblotting were utilized to confirm functional mutation of the DJ-1 gene. Normal DJ-1 protein expression in adult mouse tissue was characterized and demonstrates high expression in brain tissue with wide systemic distribution. Primary astrocytes isolated from DJ-1(-/-) mice reveal a decreased nuclear localization of DJ-1 protein in response to rotenone or LPS, with a concomitant increase in mitochondrial localization of DJ-1 found only in the rotenone exposure. Resting mitochondrial membrane potential was significantly lower in DJ-1(-/-) astrocytes, as compared to controls. Our DJ-1 knockout mouse provides an exciting tool for exploring the molecular and physiological roles of DJ-1 to further explicate its functions in neurodegeneration.
authors.affiliation: Cell and Molecular Biology Graduate Program, Colorado State University, Fort Collins, CO 80523, United States.
authors.forename: Amanda K
authors.initials: AK
authors.lastname: Ashley
authors.forename: William H
authors.initials: WH
authors.lastname: Hanneman
authors.forename: Takeshi
authors.initials: T
authors.lastname: Katoh
authors.forename: Julie A
authors.initials: JA
authors.lastname: Moreno
authors.forename: Ashley
authors.initials: A
authors.lastname: Pollack
authors.forename: Ronald B
authors.initials: RB
authors.lastname: Tjalkens
authors.forename: Marie E
authors.initials: ME
authors.lastname: Legare
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005838:Genotype; D008070:Lipopolysaccharides / Q000494:pharmacology; D053078:Membrane Potential, Mitochondrial; D051379:Mice; D018345:Mice, Knockout; D008928:Mitochondria / Q000378:metabolism; D009154:Mutation; D015513:Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D054464:Peroxiredoxins; D010641:Phenotype; D000071617:Protein Deglycase DJ-1; D021381:Protein Transport; D012402:Rotenone / Q000494:pharmacology
chemical_list: D008070:Lipopolysaccharides; D015513:Oncogene Proteins; D012402:Rotenone; D054464:Peroxiredoxins; C503466:PARK7 protein, mouse; D000071617:Protein Deglycase DJ-1