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. 2012 Jan 16;13:1. doi: 10.1186/1471-2199-13-1

Figure 1.

Figure 1

Confocal images of DSB signals generated by plant SPO11 expression in the mei-W68 mus301 double mutant background (mei-W681/mei-W681; mus301D4/Df(3L)66C-G28). The DNA is green. The γ-H2Av is red, and shows the DSB signals. When the two signals overlap, the signal is yellow. Each inset shows an enlarged view of the oocyte nucleus. All images are single confocal sections. Each scale bar shows 20 μm. (A) An egg chamber of mei-W68+ /mei-W681; mus301D4/Df(3L)66C-G28. (B) An egg chamber of mei-W681/mei-W681; mus301D4/Df(3L)66C-G28. The frequencies of the oocyte nuclei with DSB signals for the total oocyte nuclei of stage 2-8 egg chambers were 100% (344/345, 3 ovaries) in the mei-W681 heterozygote, and 7.4% (16/220, 2 ovaries) in the mei-W681 homozygote (see Figure 2A). The frequencies of the oocyte nuclei with a karyosome morphological defect (Figure 1) for the total oocyte nuclei of stage 3-8 egg chambers were 91% (254/278, 3 ovaries) in the mei-W68 heterozygote, and 1.3% (2/177, 2 ovaries) in the mei-W68 homozygote (see Figure 2B). (C) An egg chamber of mei-W681 /mei-W681; mus301D4 /Df(3L)66C-G28 with transgene P{hsp83-mei-W68+cDNA, M53-3} (indicated as Vector name{gene, insertion #}). The frequency of the oocyte nuclei with DSB signals for the total oocyte nuclei of stage 2-8 egg chambers was 100% (180/180, 2 ovaries) in the mei-W68 homozygote with P{hsp83-mei-W68+cDNA, M53-3}, similar to the mei-W681 heterozygote. The frequency of the oocyte nuclei with defective karyosome morphology for the total oocyte nuclei of stage 3-8 egg chambers was 70% (93/132) in the mei-W68 homozygote with P{hsp83-mei-W68+cDNA, M53-3}, similar to the mei-W681 heterozygote. The karyosomes in the insets of panels A and C (SPO11 positive oocyte with DSB repair deficiency, as positive controls) have defective morphology (non-disc shape), while the karyosome in the inset of panel B (DSB-repair defective oocyte without functional SPO11 as negative controls) has normal morphology (disc shape). (D-G) Egg chambers of mei-W681 /mei-W681; mus301D4 /Df(3L)66C-G28 with transgene: P{hsp83-AtSPO11-1 cDNA, A5-1} (D); P{hsp83-AtSPO11-2 cDNA, 2M3-1} (E); P{hsp83-OsSPO11A cDNA, 3M2-3} (F); P{hsp83-OsSPO11D cDNA, F2-1} (G). Quantitative data with respect to the DSB signals and the defective karyosome morphology in the oocyte nuclei of these transgenic flies are shown in Figure 2.