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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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Observation of maternal expression and post-fertilization translocation of Apo-14 promoter-driven GFP in transgenic zebrafish line Tg(Apo14: GFP).(a) unfertilized egg; (b) 1-cell embryo; (c) 2-cell embryo; (d) high blastula embryo; (e) 50%-epiboly embryo; (f) shield embryo; (g) 90%-epiboly embryo; (h) bud stage embryo.
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pone-0022555-g003: Observation of maternal expression and post-fertilization translocation of Apo-14 promoter-driven GFP in transgenic zebrafish line Tg(Apo14: GFP).(a) unfertilized egg; (b) 1-cell embryo; (c) 2-cell embryo; (d) high blastula embryo; (e) 50%-epiboly embryo; (f) shield embryo; (g) 90%-epiboly embryo; (h) bud stage embryo.

Mentions: Identical to the presence of maternal Apo-14 in gibel carp [16], the Apo-14 promoter-driven GFP was also expressed in mature eggs of the Tg(Apo14: GFP) zebrafish, and uniformly distributed in the yolk (Figure3a). After the egg was fertilized, the GFP moved to animal pole along with the cytoplasm stream (Figure3b). Following the cleavage progress, the maternal GFP was equally distributed to the blastomeres (Figure3c–g), and gradually disappeared until the bud stage (Figure3h).


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

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

Observation of maternal expression and post-fertilization translocation of Apo-14 promoter-driven GFP in transgenic zebrafish line Tg(Apo14: GFP).(a) unfertilized egg; (b) 1-cell embryo; (c) 2-cell embryo; (d) high blastula embryo; (e) 50%-epiboly embryo; (f) shield embryo; (g) 90%-epiboly embryo; (h) bud stage embryo.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0022555-g003: Observation of maternal expression and post-fertilization translocation of Apo-14 promoter-driven GFP in transgenic zebrafish line Tg(Apo14: GFP).(a) unfertilized egg; (b) 1-cell embryo; (c) 2-cell embryo; (d) high blastula embryo; (e) 50%-epiboly embryo; (f) shield embryo; (g) 90%-epiboly embryo; (h) bud stage embryo.
Mentions: Identical to the presence of maternal Apo-14 in gibel carp [16], the Apo-14 promoter-driven GFP was also expressed in mature eggs of the Tg(Apo14: GFP) zebrafish, and uniformly distributed in the yolk (Figure3a). After the egg was fertilized, the GFP moved to animal pole along with the cytoplasm stream (Figure3b). Following the cleavage progress, the maternal GFP was equally distributed to the blastomeres (Figure3c–g), and gradually disappeared until the bud stage (Figure3h).

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