IMAGE

Fig. 2

ID
ZDB-IMAGE-130830-2
Genes
Antibodies
Source
Figures for Ninov et al., 2013
Image
Figure Caption

Fig. 2

Nutrients Regulate β Cell Differentiation (A) Tg(TP1:CreERT2);Tg(ubi3C:loxp-GFP-loxp:mCherry) larvae were treated with 4-OHT at 14 dpf for 16 hr, fixed at 17 dpf, and stained for 2F11 to mark IPD cells. 4-OHT treatment resulted in the mosaic labeling of individual IPD cells (mCherry+;GFP cells) (arrowheads). (B) Experimental setup for the lineage tracing of β cells from IPD cells. Tg(insulin:loxP:mCherry-STOP:loxP:H2BGFP)+ β cells that originated from IPD cells with Tg(TP1:CreERT2) activity exhibit H2BGFP expression instead of mCherry expression. In a single-progenitor scenario (1), each SI would be composed of H2BGFP+ or mCherry+ cells, whereas, in a multiple-progenitor scenario (2), SIs would be mosaic, containing both H2BGFP+ and mCherry+ cells. (C–F) Tg(TP1:CreERT2);Tg(insulin:loxP:mCherry-STOP:loxP:H2BGFP) larvae treated with limiting concentrations of 4-OHT at 16 dpf for 16 hr and analyzed at 40 dpf. (C) A mosaic SI composed of H2BGFP+ (arrowheads) and mCherry+ β cells. (D) Three individual H2BGFP+ β cells (arrowheads) located in the periphery of a SI, suggesting that they originated from three different IPD cells. (E) A PI showing two peripheral H2BGFP+ groups of β cells (arrowheads), indicating several cell cycles after differentiation. (F) A single H2BGFP+ β cell (arrowhead) in proximity to an SI. Of 20 animals treated with 4-OHT, 15 contained lineage-traced cells, whereas no lineage-traced cells were observed in vehicle-treated controls (n = 7 animals, 80 SIs and 7 PIs). (G) Tg(TP1:H2BmCherry);Tg(ins:GFP) animals were examined at 30 dpf. A projection of several planes shows an SI (arrow). The white arrowhead points to a single β cell in the periphery of the SI. The yellow arrowhead points to a single β cell outside the SI, extending a long cellular process toward it. Both of these β cells exhibit higher levels of Tg(TP1:H2BmCherry) fluorescence in comparison to the rest of the β cells in the SI, suggesting that they recently differentiated and did not undergo proliferation. (H and I) Tg(ins:Kaede) animals were maintained on a low-calorie diet until 20 dpf (H) or switched to a high-calorie diet (I) from 15 to 20 dpf (see Experimental Procedures). Although there is only a single Tg(ins:Kaede)+ SI (arrowhead) posterior to the PI (arrow) in (H), multiple SIs (arrowheads) formed in (I). (J) Quantification of the number of SIs posterior to the PI (n = 17 animals for each group). The high-calorie diet induced the formation of more SIs in comparison to the low-calorie diet (****p < 0.0001). Error bars represent SEM, and scale bars represent 20 μm. See also Figure S2.

Figure Data
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Curr. Biol.