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Related Experiment Videos

Improved fragile site detection with trimethoprim.

D A McLean1, M J Faed

  • 1Department of Pathology, University of Dundee, Ninewells Hospital, UK.

Human Genetics
|July 1, 1990
PubMed
Summary

Trimethoprim, a dihydrofolate reductase inhibitor, effectively reveals fragile sites on human chromosomes. Adding trimethoprim early in lymphocyte cultures significantly increased fragile site frequencies compared to other methods.

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Area of Science:

  • Human genetics
  • Molecular biology
  • Cytogenetics

Background:

  • Fragile sites are specific chromosomal regions prone to breakage.
  • Identifying fragile sites is crucial for understanding genomic instability and associated diseases.
  • Dihydrofolate reductase (DHFR) is a key enzyme in folate metabolism, essential for DNA synthesis.

Purpose of the Study:

  • To investigate the efficacy of trimethoprim as a chemical agent for inducing and visualizing fragile sites on human chromosomes.
  • To determine the optimal conditions for trimethoprim-induced fragile site expression in lymphocyte cultures.

Main Methods:

  • Human peripheral blood lymphocytes were cultured under various conditions.
  • Trimethoprim, a DHFR inhibitor, was added at different time points during lymphocyte culture.
  • Chromosomal analysis was performed to quantify the frequency of fragile sites.

Main Results:

  • Lymphocyte cultures treated with 20 mg/l trimethoprim at the onset of incubation exhibited significantly higher frequencies of fragile sites.
  • Compared to other culture regimes and timings, early trimethoprim addition proved most effective in demonstrating fragile sites.
  • The study confirmed trimethoprim's role in enhancing the visibility of chromosomal fragile sites.

Conclusions:

  • Trimethoprim is a potent inducer of fragile sites in human chromosomes.
  • Early addition of trimethoprim to lymphocyte cultures is a reliable method for increasing fragile site detection.
  • This finding aids in the cytogenetic analysis of chromosomal instability and related genetic disorders.

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