Analysis of the Ambient Particulate Matter-Induced Chromosomal Aberrations Using an In Vitro System

Details

• Personal Author:
  Chalbot M-C ; Hauer-Jensen M ; Kavouras I ; Koturbash I ; Miousse IR ; Pathak R
• Description:
  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. [Description provided by NIOSH]
• Subjects:
  Air Pollution Blood Cells, Cultured Chromosomes Cytotoxins Environmental Monitoring Genetics Laboratories Macrophages Occupational Health Particulate Matter Safety Spectrum Analysis

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• Keywords:
  Air Pollution Air Sampling Author Keywords: Ambient Particulate Matter Cell Alteration Cell Cultures Cell Culture Techniques Cell Cycle Cellular Effects Chromosomal Aberrations Chromosome Damage Chromosome Disorders Cytogenetic Cytotoxic Effects DNA Damage Exposure Assessment Gene Mutation Genotoxic Effects In Vitro In Vitro Study Laboratory Animals Magnetic Resonance Imaging Mitosis MRI Particulate Matter Collection Particulates Spectrographic Analysis

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• ISSN:
  1940-087X
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  Alabama ; Arkansas ; OSHA Region 4 ; OSHA Region 6
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Issue:
  118
• NIOSHTIC Number:
  nn:20051553
• Citation:
  J Vis Exp 2016 Dec; (118):e54969
• Contact Point Address:
  Isabelle R. Miousse, Department of Occupational and Environmental Health, University of Arkansas for Medical Sciences
• Email:
  IRacinemiousse@uams.edu
• Federal Fiscal Year:
  2017
• Performing Organization:
  University of Alabama at Birmingham
• Peer Reviewed:
  True
• Start Date:
  20050701
• Source Full Name:
  Journal of Visualized Experiments
• End Date:
  20270630
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:bdf7c10da6817323f27dded58f4083642fa61b3eaa2beb303324b1db836011d360857e8680735ce625bd8c6784e0b735de6057b86245529a6289db0b9223ec3a
• Download URL:
  https://stacks.cdc.gov/view/cdc/211450/cdc_211450_DS1.pdf
• File Type:
  [PDF - 725.31 KB ]