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An Apo-14 promoter-driven transgenic zebrafish that marks liver organogenesis.

Wang R, Li Z, Wang Y, Gui JF - PLoS ONE (2011)

Bottom Line: Then, a green fluorescence dot begins to appear between the notochord and the yolk sac adjacent to otic vesicle at about 20 hpf, which is later demonstrated to be liver primordium that gives rise to liver.Additionally, we observed similar morphology between the liver progenitor cells and the GFP-positive nuclei on the YSL, suggesting that they might originate from the same progenitor cells in early embryos.Overall, the current study provides a transgenic zebrafish line that marks the through liver organogenesis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China.

ABSTRACT
Several transgenic zebrafish lines for liver development studies had been obtained in the first decade of this century, but not any transgenic GFP zebrafish lines that mark the through liver development and organogenesis were reported. In this study, we analyzed expression pattern of endogenous Apo-14 in zebrafish embryogenesis by whole-mount in situ hybridization, and revealed its expression in liver primordium and in the following liver development. Subsequently, we isolated zebrafish Apo-14 promoter of 1763 bp 5'-flanking sequence, and developed an Apo-14 promoter-driven transgenic zebrafish Tg(Apo14: GFP). And, maternal expression and post-fertilization translocation of Apo-14 promoter-driven GFP were observed in the transgenic zebrafish line. Moreover, we traced onset expression of Apo-14 promoter-driven GFP and developmental behavior of the expressed cells in early heterozygous embryos by out-crossing the Tg(Apo14: GFP) male to the wild type female. Significantly, the Apo-14 promoter-driven GFP is initially expressed around YSL beneath the embryo body at 10 hpf when the embryos develop to tail bud prominence. In about 14-somite embryos at 16-17 hpf, a typical "salt-and-pepper" expression pattern is clearly observed in YSL around the yolk sac. Then, a green fluorescence dot begins to appear between the notochord and the yolk sac adjacent to otic vesicle at about 20 hpf, which is later demonstrated to be liver primordium that gives rise to liver. Furthermore, we investigated dynamic progression of liver organogenesis in the Tg(Apo14: GFP) zebrafish, because the Apo-14 promoter-driven GFP is sustainably expressed from hepatoblasts and liver progenitor cells in liver primordium to hepatocytes in the larval and adult liver. Additionally, we observed similar morphology between the liver progenitor cells and the GFP-positive nuclei on the YSL, suggesting that they might originate from the same progenitor cells in early embryos. Overall, the current study provides a transgenic zebrafish line that marks the through liver organogenesis.

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Onset expression and developmental behavior of Apo-14 promoter-driven GFP in early heterozygous embryos produced by out-crossing the Tg(Apo14: GFP) male to the wild type female.(a, a’) bud embryo at 10 hpf; (b, b’) 5-somite embryo at 12 hpf; (c, c’) 14-somite embryo at 16 hpf; (d, d’) 17 hpf embryo; (e, e’) 20 hpf embryo; (f, f’) 24 hpf embryo. Arrows point to the liver primordium. The images (a–f) show the green fluorescence, and (a’–f’) the corresponding bright images.
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pone-0022555-g004: Onset expression and developmental behavior of Apo-14 promoter-driven GFP in early heterozygous embryos produced by out-crossing the Tg(Apo14: GFP) male to the wild type female.(a, a’) bud embryo at 10 hpf; (b, b’) 5-somite embryo at 12 hpf; (c, c’) 14-somite embryo at 16 hpf; (d, d’) 17 hpf embryo; (e, e’) 20 hpf embryo; (f, f’) 24 hpf embryo. Arrows point to the liver primordium. The images (a–f) show the green fluorescence, and (a’–f’) the corresponding bright images.

Mentions: To clarify the onset expression of Apo-14 promoter-driven GFP and thereby to trace the developmental behavior of the expressed cells in early embryos, we produced heterozygous embryos by out-crossing the Tg(Apo14: GFP) male to the wild type female, because the heterozygous embryos avoided maternal GFP, and were able to characterize the onset and progression of expression with confocal laser scanning microscope. As shown in Figure4, the earliest GFP fluorescence is initially observed around YSL beneath the embryo body at 10 hpf when the embryos develop to tail bud prominent (Figure4a), and the green fluorescence ring becomes obvious at 12 hpf when the embryos develop to 5-somite stage (Figure4b). In about 14-somite embryos at 16–17 hpf, a typical “salt-and-pepper” expression pattern is clearly observed in YSL around the yolk sac (Figure4c and 4d), and green fluorescence also appears in the notochord at 17 hpf. In comparison with the corresponding bright images (Figure4c’ and 4d’), the GFP expressed cells locate in YSL with an ordered distribution. At about 20 hpf, a green fluorescence dot begins to appear between the notochord and the yolk sac adjacent to otic vesicle (Figure4e and 4e’). Significantly, the dot exists continually as observed at 24 hpf (Figure4f), and becomes very obvious along with the following developmental progress. Additionally, eyes are also lightened by the Apo-14 promoter-driven GFP at about 24 hpf.


An Apo-14 promoter-driven transgenic zebrafish that marks liver organogenesis.

Wang R, Li Z, Wang Y, Gui JF - PLoS ONE (2011)

Onset expression and developmental behavior of Apo-14 promoter-driven GFP in early heterozygous embryos produced by out-crossing the Tg(Apo14: GFP) male to the wild type female.(a, a’) bud embryo at 10 hpf; (b, b’) 5-somite embryo at 12 hpf; (c, c’) 14-somite embryo at 16 hpf; (d, d’) 17 hpf embryo; (e, e’) 20 hpf embryo; (f, f’) 24 hpf embryo. Arrows point to the liver primordium. The images (a–f) show the green fluorescence, and (a’–f’) the corresponding bright images.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3142191&req=5

pone-0022555-g004: Onset expression and developmental behavior of Apo-14 promoter-driven GFP in early heterozygous embryos produced by out-crossing the Tg(Apo14: GFP) male to the wild type female.(a, a’) bud embryo at 10 hpf; (b, b’) 5-somite embryo at 12 hpf; (c, c’) 14-somite embryo at 16 hpf; (d, d’) 17 hpf embryo; (e, e’) 20 hpf embryo; (f, f’) 24 hpf embryo. Arrows point to the liver primordium. The images (a–f) show the green fluorescence, and (a’–f’) the corresponding bright images.
Mentions: To clarify the onset expression of Apo-14 promoter-driven GFP and thereby to trace the developmental behavior of the expressed cells in early embryos, we produced heterozygous embryos by out-crossing the Tg(Apo14: GFP) male to the wild type female, because the heterozygous embryos avoided maternal GFP, and were able to characterize the onset and progression of expression with confocal laser scanning microscope. As shown in Figure4, the earliest GFP fluorescence is initially observed around YSL beneath the embryo body at 10 hpf when the embryos develop to tail bud prominent (Figure4a), and the green fluorescence ring becomes obvious at 12 hpf when the embryos develop to 5-somite stage (Figure4b). In about 14-somite embryos at 16–17 hpf, a typical “salt-and-pepper” expression pattern is clearly observed in YSL around the yolk sac (Figure4c and 4d), and green fluorescence also appears in the notochord at 17 hpf. In comparison with the corresponding bright images (Figure4c’ and 4d’), the GFP expressed cells locate in YSL with an ordered distribution. At about 20 hpf, a green fluorescence dot begins to appear between the notochord and the yolk sac adjacent to otic vesicle (Figure4e and 4e’). Significantly, the dot exists continually as observed at 24 hpf (Figure4f), and becomes very obvious along with the following developmental progress. Additionally, eyes are also lightened by the Apo-14 promoter-driven GFP at about 24 hpf.

Bottom Line: Then, a green fluorescence dot begins to appear between the notochord and the yolk sac adjacent to otic vesicle at about 20 hpf, which is later demonstrated to be liver primordium that gives rise to liver.Additionally, we observed similar morphology between the liver progenitor cells and the GFP-positive nuclei on the YSL, suggesting that they might originate from the same progenitor cells in early embryos.Overall, the current study provides a transgenic zebrafish line that marks the through liver organogenesis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China.

ABSTRACT
Several transgenic zebrafish lines for liver development studies had been obtained in the first decade of this century, but not any transgenic GFP zebrafish lines that mark the through liver development and organogenesis were reported. In this study, we analyzed expression pattern of endogenous Apo-14 in zebrafish embryogenesis by whole-mount in situ hybridization, and revealed its expression in liver primordium and in the following liver development. Subsequently, we isolated zebrafish Apo-14 promoter of 1763 bp 5'-flanking sequence, and developed an Apo-14 promoter-driven transgenic zebrafish Tg(Apo14: GFP). And, maternal expression and post-fertilization translocation of Apo-14 promoter-driven GFP were observed in the transgenic zebrafish line. Moreover, we traced onset expression of Apo-14 promoter-driven GFP and developmental behavior of the expressed cells in early heterozygous embryos by out-crossing the Tg(Apo14: GFP) male to the wild type female. Significantly, the Apo-14 promoter-driven GFP is initially expressed around YSL beneath the embryo body at 10 hpf when the embryos develop to tail bud prominence. In about 14-somite embryos at 16-17 hpf, a typical "salt-and-pepper" expression pattern is clearly observed in YSL around the yolk sac. Then, a green fluorescence dot begins to appear between the notochord and the yolk sac adjacent to otic vesicle at about 20 hpf, which is later demonstrated to be liver primordium that gives rise to liver. Furthermore, we investigated dynamic progression of liver organogenesis in the Tg(Apo14: GFP) zebrafish, because the Apo-14 promoter-driven GFP is sustainably expressed from hepatoblasts and liver progenitor cells in liver primordium to hepatocytes in the larval and adult liver. Additionally, we observed similar morphology between the liver progenitor cells and the GFP-positive nuclei on the YSL, suggesting that they might originate from the same progenitor cells in early embryos. Overall, the current study provides a transgenic zebrafish line that marks the through liver organogenesis.

Show MeSH
Related in: MedlinePlus