Morphological defects of ovarioles induced by high-dose irradiation.

<p>The wild type females were irradiated at 40 Gy, and the germline cells were stained with an anti-Vasa antibody (green) and DAPI (blue) each day after irradiation. (A) Various defects (indicated by an arrow) were observed in oogenesis after irradiation. Representative images exhibiting the following defects are shown: germarium lacking anti-Vasa-stained germline cells in region 3 and/or region 2b (a), in region 1 (b), or in the entire germarium (c); ovarioles lacking a couple of early stage egg chambers (j) and several late stage egg chambers (k); germarium exhibiting fusion of two cysts (d); fused egg chambers (g); single germline cells in the germarium (e); abnormal number of germline cells in the egg chambers (f); ovarioles containing smaller egg chambers at more posterior positions (g, arrowhead); egg chambers with an abnormal shape (h); and egg chambers containing mislocalized nurse cells (l). The germarium (g) and the stage of the egg chambers are shown in (j) and (k). The scale bars represent 20 µm (a–i) and 50 µm (j–l). (B) Irradiation induced loss of germline cells in the germarium. The percentage of germarium lacking anti-Vasa-stained germline cells in region 3 and/or 2 (blue dots), region 1 (red diamonds), or the entire germarium (green triangles) was counted each day after irradiation. The values represent the means and standard deviations from two independent experiments. At least 55 ovarioles were counted in total for each time point. (C) Morphological defects observed in the germarium and egg chambers. The percentage of germarium containing morphological defects in the germline cells (blue dots) or extracellular materials (green triangles) was counted. The percentage of ovarioles containing an abnormal egg chamber morphology (red diamonds) was also calculated. The values represent the means and standard deviations from two independent experiments. At least 55 ovarioles were counted in total for each time point.