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A new FISH assay to simultaneously detect structural and numerical chromosomal abnormalities in mouse sperm
Francesca S Hill1, Francesco Marchetti, Melissa Liechty
1Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California, USA.
Molecular Reproduction and Development
|September 2, 2003
Summary
Researchers developed a new fluorescence in situ hybridization (FISH) assay to detect structural chromosome abnormalities in mouse sperm. This assay reveals a lower baseline frequency of these aberrations in mice compared to humans.
Area of Science:
- Genetics
- Cytogenetics
- Toxicology
Background:
- De novo structural aberrations in chromosomes are a significant source of paternally transmitted genetic damage.
- The majority of de novo structural aberrations in human offspring originate paternally, unlike numerical abnormalities.
Purpose of the Study:
- To develop and validate a novel three-color fluorescence in situ hybridization (FISH) assay, termed CT8, for detecting mouse sperm with structural and numerical chromosomal abnormalities.
- To establish baseline frequencies of specific chromosomal aberrations in mouse sperm using the CT8 assay.
Main Methods:
- Development of a three-color FISH assay (CT8) utilizing DNA probes for centromeric and telomeric regions of chromosome 2, and a subcentromeric region probe for chromosome 8.
- Analysis of approximately 80,000 mouse sperm to quantify frequencies of structural aberrations involving chromosome 2, disomy 2, disomy 8, and sperm diploidy.
- Validation of the CT8 assay by comparing results with chromosome painting cytogenetic analysis in T(2;14) translocation carrier mice.
Main Results:
- The CT8 assay detected an average baseline frequency of 2.5 per 10,000 sperm with partial duplications and deletions of chromosome 2.
- An estimated 0.4% of mouse sperm carry structural chromosomal aberrations, which is over five times lower than spontaneous frequencies observed in humans.
- The CT8 assay demonstrated good concordance with established cytogenetic methods for detecting abnormal segregants in translocation carriers.
Conclusions:
- The CT8 sperm FISH assay is a reliable and sensitive method for detecting structural chromosome aberrations in mouse sperm.
- The developed assay provides valuable insights into the frequency of paternal chromosomal damage in mice.
- The CT8 assay has broad applicability in genetic research, reproductive physiology studies, and genetic toxicology assessments.