ZFIN Image: Ye et al., 2016, Fig. 2

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Fig. 2

Insulin signaling blockade increases embryonic β cell formation. (A) Schematic of a dominant negative IRS2 construct designed to block transmission of insulin signaling; C-terminal effector binding domains are replaced with GFP. (B,C) 5 hpf embryo injected with dnIRS2-GFP mRNA (B) Photomicrograph of epifluorescence shows ubiquitous distribution of GFP. (C) Confocal plane shows the localization of dnIRS2-GFP to the plasma membrane while co-injected H2B-RFP mRNA labels cell nuclei red. (D,E) Confocal projections of control (D) and dnIRS2-GFP mRNA-injected islets in 24 hpf Tg(ins:dsRed) embryos. (F) Quantification of insa:dsRed+ β cells in 24 hpf control (n=12) and dnIRS2-GFPmRNA injected embryos (n=10). (G,H) Confocal projections of DMSO-treated control (G) and 1 µM wortmannin-treated islets in 30 hpf Tg(ins:CFP-NTR) embryos. (I) Quantification of β cells in DMSO-treated control (n=18) and 1 µM wortmannin-treated (n=17) islets in 30 hpf embryos. Student t-test was used to determine significance in F and I.

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Acknowledgments

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Reprinted from Developmental Biology, 409(2), Ye, L., Robertson, M.A., Mastracci, T.L., Anderson, R.M., An insulin signaling feedback loop regulates pancreas progenitor cell differentiation during islet development and regeneration, 354-69, Copyright (2016) with permission from Elsevier. Full text @ Dev. Biol.