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Cytopenia induction by 5-fluorouracil identifies thrombopoietic mutants in sensitized ENU mutagenesis screens. | LitMetric

AI Article Synopsis

  • * Researchers optimized the 5FU dose across different strains of mice and performed N-ethyl-N-nitrosourea (ENU) mutagenesis on one strain to identify phenotypic variants related to platelet recovery.
  • * Out of 546 tested mice, 15 showed significant changes in platelet number and recovery, demonstrating that the combined use of 5FU and specific mouse strains can effectively uncover mutations related to blood cell development.

Article Abstract

The ability of random mutagenesis techniques to annotate the mammalian genome can be hampered due to genetic redundancy and compensatory pathways that mask heterozygous mutations under homeostatic conditions. The objective of this study was to devise a pharmacologically sensitized screen using the chemotherapeutic drug, 5-fluorouracil (5FU), to induce cytopenia. 5FU dose was optimized in the 129/SvImJ, C57BL/6J, BALB/cJ, and C3H/HeJ strains of laboratory mice. N-ethyl-N-nitrosourea (ENU) mutagenesis was performed on 129/SvImJ males and phenotypic variants were identified by backcrossing on to the C57BL/6J background. G1 animals were challenged with 100 μg/g 5FU and phenodeviants with altered platelet recovery were monitored. Of 546 G1 animals tested, 15 phenodeviants were identified that displayed increased baseline platelet number, a platelet overshoot, or delayed platelet recovery, thereby demonstrating the utility of this approach for uncovering mutations in megakaryocyte and platelet development. Four G1 mice were selected for further analysis. The phenotypes were heritable in all four strains and genetic mapping identified a chromosome location in two of the three G2 lines tested. In conclusion, our group has developed a sensitized random mutagenesis screen utilizing 5FU and has shown that the strain combination of 129/SvImJ × C57BL/6J is robust for identification of founder lines with defects in megakaryocyte and platelet development.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3341667PMC
http://dx.doi.org/10.1016/j.exphem.2011.09.007DOI Listing

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