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When cell division is blocked in the four-cell embryo with cytochalasin B the nuclear division cycle continues. The rapid S phase typical of early embryonic cycles creates four multi-nucleate cells with large amounts of DNA after only one hour of continued incubation. This assay can be combined with RNAi to evaluate the role of candidate DNA replication proteins in DNA synthesis. Shown below is a control embryo (top) and an embryo depleted of the replication protein MUS-101 (bottom) stained with DAPI to visualize DNA. Note the differences in the amount of DNA synthesized during the one hour incubation period. Note also that the middle right set of nuclei in the wild type embryo are in mitosis. Image by Toni Holway. |
