Meiotic checkpoint and defective eggshell induced by IR treatment.

<p>(A–B) Karyosome defects after IR treatment. Wild type females were irradiated at 40 Gy, and the ovaries were stained for DNA with propidium iodide (red) and for oocyte cytoplasm with an anti-Orb antibody (green). (A) Representative confocal images of stage 6 egg chambers before (a) and after irradiation (b, c, and d) are shown. Bar, 10 µm. The nuclei of oocytes are indicated with a white arrow. (B) The percentage of oocytes at stage 6 with defective karyosome was counted at 0, 1, 8, 24, 48, and 72 h after irradiation. The morphology of the karyosome similar to (Ad) was considered as defective. The data are presented as the mean and standard deviation of at least two independent experiments. The total number of oocytes at each time point was at least 21. (C–D) Dorsal-ventral patterning defects of the embryos laid by the irradiated females. (C) Representative images of a normal embryo from an untreated female (-IR) and a defective embryo from a wild type female irradiated at 40 Gy (+IR) are shown. (D) Wild type or <i>mnk</i> mutant females were irradiated at 40 Gy and the embryos were collected every 24 h. The percentage of embryos with dorsal-ventral patterning defects were counted each day. Day 1 corresponds to the embryos collected 0–24 h after irradiation. The data are presented as the mean and standard deviation of at least two independent experiments. At least 100 embryos were observed for each time point in each experiment. (E–F) Eggshell defects in the embryos derived from irradiated females. (E) The embryos derived from the untreated (-IR, normal eggshell) and 40 Gy irradiated (+IR) females are shown. The embryos from irradiated female exhibited irregular eggshells (second panel, indicated in the red dotted line) or lucid eggshells (third panel). (F) The embryos were collected from the irradiated females every 24 h, and the percentages of the embryos with thin eggshell phenotypes were counted. Day 1 represents the embryos collected at the first 24 h after irradiation. The data are presented as the mean and standard deviation of three independent experiments. At least 100 embryos were observed for each time point.