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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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Samples were hybridised with Api-piwi1,4,7 (A–F) and Api-piwi8 (G–L) antisense riboprobes. Anterior is to the left; dorsal is upper. All views are lateral. Unless addressed specifically, locations of germ cells are labelled with double arrowheads. (A, G) Germaria and segregated oocytes (stage 1). Preferential expression was identified in the follicle cells between the germarium and the oocyte (arrowheads). Api-piwi8 transcripts were not detected. (B, H) Embryos undergoing nuclear divisions (stage 2, 3). Api-piwi1,4,7 expression remained restricted to the follicle cells (arrowheads). Api-piwi8 stayed undetected. (C, I) Embryos with newly-segregated germ cells (stage 6). Most Api-piwi1,4,7 transcripts were restricted to the central syncytium (arrow), but Api-piwi1,4,7 expression was not detected in germ cells. Api-piwi8 was weakly expressed in anterior two thirds of the egg. (D, J) Embryos undergoing gastrulation (stage 8). Api-piwi1,4,7 expression was preferentially identified in the invaginating germband (arrow). Api-piwi8 expression was almost undetected. (E, K) Extension of the germband (stage 12). Strong expression of Api-piwi1,4,7 was identified in the abdomen (arrow). Faint expression of Api-piwi8 was visible. (F, L) Completion of germband retraction (stage 18). Api-piwi1,4,7 transcripts were restricted to the gonadal germ cells (arrow); Api-piwi8 expression was almost undetected. (M, N) Negative control, ovarioles hybridised with sense riboprobes. No signals were detected. (O) Illustration displaying presented developmental stages. Abbreviations: Ab, abdomen; Ant, antennae; Bc, endosymbiotic bacteria (invading embryos during stage 7); Blc, blastodermal cells; CNS; central nerve system; Cs, central syncytium (central blastoderm containing multiplying nuclei); Dev, development; Dn, dividing nuclei; Fc, follicle cells; Gc, germ cells; Gl, germarial lumen (central cavity of the germarium, where germline stem cells are derived); Gm, germarium; Hd, head; Ig, invaginating germband; L1-3, limbs 1–3; Nc, nurse cells; Nu, nuclei; Oc, oocyte; Ps, posterior syncytium; Sn, syncytial nuclei. St, stage; Th, thorax; T1-3, thoracic segments 1–3. Scale bar, 20 µm.