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High-throughput Screening and Biosensing with Fluorescent C. elegans Strains
Published on: May 19, 2011
A genetic screen for temperature-sensitive cell-division mutants of Caenorhabditis elegans
K F O'Connell1, C M Leys, J G White
1Laboratory of Molecular Biology, University of Wisconsin, Madison, Wisconsin 53706, USA. kfoconne@facstaff.wisc.edu
Abstract:
A novel screen to isolate conditional cell-division mutants in Caenorhabditis elegans has been developed. The screen is based on the phenotypes associated with existing cell-division mutations: some disrupt postembryonic divisions and affect formation of the gonad and ventral nerve cord-resulting in sterile, uncoordinated animals-while others affect embryonic divisions and result in lethality. We obtained 19 conditional mutants that displayed these phenotypes when shifted to the restrictive temperature at the appropriate developmental stage. Eighteen of these mutations have been mapped; 17 proved to be single alleles of newly identified genes, while 1 proved to be an allele of a previously identified gene. Genetic tests on the embryonic lethal phenotypes indicated that for 13 genes, embryogenesis required maternal expression, while for 6, zygotic expression could suffice. In all cases, maternal expression of wild-type activity was found to be largely sufficient for embryogenesis. Cytological analysis revealed that 10 mutants possessed embryonic cell-division defects, including failure to properly segregate DNA, failure to assemble a mitotic spindle, late cytokinesis defects, prolonged cell cycles, and improperly oriented mitotic spindles. We conclude that this approach can be used to identify mutations that affect various aspects of the cell-division cycle.
Insights
Researchers developed a new method to find conditional cell-division mutants in Caenorhabditis elegans. This screen identified 19 mutants affecting cell division, providing insights into essential genes for development.
Area of Science:
- Developmental Biology
- Genetics
- Cell Biology
Background:
- Cell division is fundamental to development and organismal integrity.
- Conditional mutants are crucial for studying essential genes that are lethal under standard conditions.
- Caenorhabditis elegans is a powerful model organism for genetic screens due to its rapid life cycle and conserved biology.
Purpose of the Study:
- To develop and implement a novel genetic screen for isolating conditional cell-division mutants in Caenorhabditis elegans.
- To identify new genes involved in regulating cell division during different developmental stages.
- To characterize the roles of maternal and zygotic gene expression in embryogenesis.
Main Methods:
- A conditional screen was designed based on known cell-division mutant phenotypes (sterility, lethality).
- Mutants were identified by temperature shifts to a restrictive condition at specific developmental stages.
- Genetic mapping, allelism tests, and cytological analyses were performed on isolated mutants.
Main Results:
- 19 conditional cell-division mutants were isolated, displaying phenotypes affecting postembryonic or embryonic divisions.
- 18 mutations were mapped to 17 newly identified genes and 1 known gene.
- Cytological analysis revealed diverse defects in 10 mutants, including DNA segregation, spindle assembly, and cytokinesis.
Conclusions:
- The developed screen is effective for identifying conditional cell-division mutants in Caenorhabditis elegans.
- This approach has uncovered novel genes essential for cell division and development.
- The findings provide a foundation for further research into the molecular mechanisms governing the cell-division cycle.
