title: Analysis of the Ambient Particulate Matter-induced Chromosomal Aberrations Using an In Vitro System.
abstract: Exposure to particulate matter (PM) is a major world health concern, which may damage various cellular components, including the nuclear genetic material. To assess the impact of PM on nuclear genetic integrity, structural chromosomal aberrations are scored in the metaphase spreads of mouse RAW264.7 macrophage cells. PM is collected from ambient air with a high volume total suspended particles sampler. The collected material is solubilized and filtered to retain the water-soluble, fine portion. The particles are characterized for chemical composition by nuclear magnetic resonance (NMR) spectroscopy. Different concentrations of particle suspension are added onto an in vitro culture of RAW264.7 mouse macrophages for a total exposure time of 72 hr, along with untreated control cells. At the end of exposure, the culture is treated with colcemid to arrest cells in metaphase. Cells are then harvested, treated with hypotonic solution, fixed in acetomethanol, dropped onto glass slides and finally stained with Giemsa solution. Slides are examined to assess the structural chromosomal aberrations (CAs) in metaphase spreads at 1,000X magnification using a bright-field microscope. 50 to 100 metaphase spread are scored for each treatment group. This technique is adapted for the detection of structural chromosomal aberrations (CAs), such as chromatid-type breaks, chromatid-type exchanges, acentric fragments, dicentric and ring chromosomes, double minutes, endoreduplication, and Robertsonian translocations in vitro after exposure to PM. It is a powerful method to associate a well-established cytogenetic endpoint to epigenetic alterations.
authors.affiliation: Department of Occupational and Environmental Health, University of Arkansas for Medical Sciences; IRacinemiousse@uams.edu.
authors.forename: Isabelle R
authors.initials: IR
authors.lastname: Miousse
authors.affiliation: Department of Occupational and Environmental Health, University of Arkansas for Medical Sciences; ikoturbash@uams.edu.
authors.forename: Igor
authors.initials: I
authors.lastname: Koturbash
authors.affiliation: Department of Environmental Health Sciences, University of Alabama at Birmingham.
authors.forename: Marie-Cécile
authors.initials: MC
authors.lastname: Chalbot
authors.affiliation: Division of Radiation Health, University of Arkansas for Medical Sciences.
authors.forename: Martin
authors.initials: M
authors.lastname: Hauer-Jensen
authors.affiliation: Department of Environmental Health Sciences, University of Alabama at Birmingham.
authors.forename: Ilias
authors.initials: I
authors.lastname: Kavouras
authors.affiliation: Division of Radiation Health, University of Arkansas for Medical Sciences.
authors.forename: Rupak
authors.initials: R
authors.lastname: Pathak
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002869:Chromosome Aberrations / Q000139:chemically induced*; D044127:Epigenesis, Genetic; D008677:Metaphase; D051379:Mice; D052638:Particulate Matter / Q000009:adverse effects*; D000067996:RAW 264.7 Cells
chemical_list: D052638:Particulate Matter