nanos3 is required cell-autonomously in the <20 μm germ cell population. (A–L) Genetic germline chimeras were created by transplanting Tg(ziwi:EGFP) donor germ cells into EGFP-negative hosts, and the contribution of EGFP+ germ cells was analyzed at 60 dpf. (A, E and I) Low magnification images, where donor derived cells are green. All others panels are higher magnification confocal images. (A–D) In controls, wild-type Tg(ziwi:EGFP) germ cells were able to equally contribute to the <20 μm germ cell population as wild-type host germ cells (WT′WT, n=10). (E-H) Wild-type Tg(ziwi:EGFP) germ cells are the only <20 μm germ cells detected in a nanos3 mutant host ovary (WT′MUT, n=5). (I–L) Conversely, nanos3 mutant; Tg(ziwi:EGFP) donor germ cells are not able to contribute to the <20 μm germ cell population in wild-type host ovaries (MUT′WT, n=2). B, F and J show merged images of EGFP expressing donor cells (green), Vasa expressing germ cells (red), and DNA (blue). C, G, and K, show endogenous GFP expression. D, H and L show Vasa protein localization, and inset shows the DNA morphology of DAPI stained cells boxed in main image. All images are from whole mount preparations. Scale bars: 200 μm (A, E and I), 20 μm (B, F and J). Stage Ia oocytes (Ia), stage Ib oocytes (Ib) and oogonia (oo).
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Reprinted from Developmental Biology, 374(2), Beer, R.L., and Draper, B.W., nanos3 maintains germline stem cells and expression of the conserved germline stem cell gene nanos2 in the zebrafish ovary, 308-318, Copyright
(2013) with permission from Elsevier.
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