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. 2011 Dec 5;6(12):e28051. doi: 10.1371/journal.pone.0028051

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Lu et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Dissected ovarioles were hybridised with DIG-labelled antisense riboprobes of Api-piwi2 (A–E) and Api-piwi5 (F–J), respectively. Orientations and morphological characteristics of embryos refer to Figure 3O. Early development: A, B, F, G; Mid development: C, D, H, I; Late development: E, J. Germ cells stained with antisense Api-piwi2 and Api-piwi5 riboprobes are highlighted with arrows (with preferential expression) and double arrows (without preferential expression). (A, F) Germaria and segregated oocytes (stage 1). Expression of Api-piwi2 was preferentially detected in the germarial lumen and the oocyte anterior (arrow heads). Api-piwi5 mRNA was expressed comparatively weaker. (B, G) Embryos with newly-segregated germ cells (stage 6). Weak expression of Api-piwi2 was specifically detected in the germ cells; expression of Api-piwi5 was evenly distributed in anterior two thirds of the embryo and it was weakly detected in the germ cells. (C, H) Gastrulating embryos (stage 8). Like stage 6, expression of Api-piwi2 was specifically detected in the germ cells but the intensity of in situ signals increased. Expression of Api-piwi5 covered most region of the embryo including germ cells. Embryo in (H) is slightly younger than that in (C) so that germ cells have not been pulled into the middle region of the embryo. Bacteria are not clearly visible in the focal plane shown. (D, I) Extension of the germband (stage 12). Transcripts of Api-piwi2 were specifically identified in the germ cells whilst transcripts of Api-piwi5 were evenly distributed except in bacteria. (E, J) Completion of germband retraction (stage 18). Specific expression of Api-piwi2 was identified in germ cells located in the dorsal region of the embryo, but universal expression of Api-piwi5 remained as it was detected in the stage-12 embryos. Abbreviations: Bc, endosymbiotic bacteria; Dev, development; Hd, head; St, stage. Scale bar, 20 µm.