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In-Vitro Chromosomal Aberration Test

Biology | Toxicology | Biochemistry | Proteomics | Pharmacology

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General Aim of Chromosomal Aberration Test

This chromosome aberration test aims at detecting structural chromosomal aberrations by estimating different classes of chromosome changes scored in metaphase using the light microscope.

 

Method of chromosome aberration test

In-Vitro screening of Metaphase Chromosomal Aberrations using Light Microscope.

Learning Objectives (ILOs)| Chromosomal Aberration Assay

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    By the end of the in vitro mammalian chromosomal aberration test, the postgraduate student will be able to: 

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    Successfully handle the required instruments and consumables needed in the chromosomal aberration test.
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    Check the confluence and count cells under the microscope.
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    Dilute the cells to a specific count suitable for seeding in the 24-well plate.
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    Calculate the concentration of tested chemicals and prepare the calculated doses in the cell culture medium.
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    Treat cells with the genotoxic agent(s) or nanoparticles and observe under the microscope during the in vitro genotoxicity testing.
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    Harvest cells, fix them and stain with Giemsa stain .
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    Analyze cells by light microscope and evaluate analyzed data.
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    Represent and interpret the resulting data graphically using dot plots.

Theoretical Background / Context

  • In genetics, genotoxicity describes the property of chemical agents that damages the genetic information within a cell causing mutations, which may lead to cancer. 
  • While genotoxicity is often confused with mutagenicity, all mutagens are genotoxic, whereas not all genotoxic substances are mutagenic. 
  • The alteration can have direct or indirect effects on the DNA: the induction of mutations mistimed event activation, and direct DNA damage leading to mutations. 
  • The permanent, heritable changes can affect either somatic cells of the organism or germ cells to be passed onto future generations. 
  • Cells prevent expression of the genotoxic mutation by either DNA repair or apoptosis; however, the damage may not always be fixed leading to mutagenesis.

Principle Work of Chromosomal Aberration Test Procedure

  • The presence of genotoxic agents in the environment may cause, through different mechanisms, chromosomal mutations and DNA aberrations, which are associated with the induction of both heritable defects, and cancer in the human population. 
  • Chromosomal mutations are distinguished under two major categories related to changes in the chromosome structure (chromosomal aberrations) and changes in the number of chromosomes (numerical aberrations). 
  • Structural chromosome aberrations are induced by physical and chemical mutagenic agents through a variety of molecular mechanisms, which are almost exclusively a consequence of a direct damage to DNA (e.g., DNA strand breaks, base damage, hydrolysis of bases, pyrimidine dimers, and DNA cross-links) or indirect (e.g., inhibition of DNA topoisomerases I and II, nucleotide pool imbalance, and generation of reactive oxygen species) left unrepaired, or misrepaired to produce chromosome breaks or rearrangements. 
  • This in vitro assay aimed to detect structural chromosomal aberrations by estimating different classes of chromosome changes scored in metaphase using the light microscope during chromosome aberration test.
  • Chromosome aberration test is based on whether one or both chromatids (at the same locus) in a metaphase chromosome are involved in an aberration in biology according to the chromosomal aberrations test procedure in chromosome aberration test.

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