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Enhancer of zeste acts as a major developmental regulator of Ciona intestinalis embryogenesis.

Le Goff E, Martinand-Mari C, Martin M, Feuillard J, Boublik Y, Godefroy N, Mangeat P, Baghdiguian S, Cavalli G - Biol Open (2015)

Bottom Line: These major phenotypic defects are specifically rescued by injection of a morpholino-resistant Ci-E(z) mRNA, which restores expression of Ci-E(z) protein and re-deposition of the H3K27me3 mark.As observed by qPCR analyses, Ci-E(z) invalidation leads to the early derepression of tissue-specific developmental genes, whereas late-acting developmental genes are generally down-regulated.Altogether, our results suggest that Ci-E(z) plays a major role during embryonic development in Ciona intestinalis by silencing early-acting developmental genes in a Hox-independent manner.

View Article: PubMed Central - PubMed

Affiliation: Université Montpellier, Place Eugène Bataillon, Montpellier 34095, Cedex 5, France Institut des Sciences de l'Evolution (ISEM), UMR5554, CNRS, Montpellier 34095, France.

No MeSH data available.


Related in: MedlinePlus

Ci-E(z) and H3K27me3 detection in rescue experiments. Comparative indirect immunofluorescence analysis between (A) control embryos, Ci-E(z) morphants, rescue embryos and (B) Ci-E(z) morphants ½ and rescue ½ embryos at late tailbud stage of development. A triple labeling was performed using: actin (Phalloïdin in green), DNA (DAPI, in blue) and antibodies against H3K27me3 or Ci-E(z) protein (in grey). On the right of each DAPI/Phalloïdin merge, the corresponding image of Ci-E(z) or H3K27me3 alone is shown (as indicated) with cell contour drawn in grey. Ab, antibodies; MO-Ctrl, control embryo; mRNA-E(z), mRNA-E(z) embryo; MO-E(z), Ci-E(z) morphant; MO-E(z) ½, embryo injected with half of MO-E(z) concentration; Rescue, mRNA-E(z)+MO-E(z) embryo; Rescue ½, embryo injected with half of MO-E(z) concentration+mRNA-E(z). For three independent experiments, between 20 and 40 embryos were collected. Scale bar: 25 µm.
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BIO010835F6: Ci-E(z) and H3K27me3 detection in rescue experiments. Comparative indirect immunofluorescence analysis between (A) control embryos, Ci-E(z) morphants, rescue embryos and (B) Ci-E(z) morphants ½ and rescue ½ embryos at late tailbud stage of development. A triple labeling was performed using: actin (Phalloïdin in green), DNA (DAPI, in blue) and antibodies against H3K27me3 or Ci-E(z) protein (in grey). On the right of each DAPI/Phalloïdin merge, the corresponding image of Ci-E(z) or H3K27me3 alone is shown (as indicated) with cell contour drawn in grey. Ab, antibodies; MO-Ctrl, control embryo; mRNA-E(z), mRNA-E(z) embryo; MO-E(z), Ci-E(z) morphant; MO-E(z) ½, embryo injected with half of MO-E(z) concentration; Rescue, mRNA-E(z)+MO-E(z) embryo; Rescue ½, embryo injected with half of MO-E(z) concentration+mRNA-E(z). For three independent experiments, between 20 and 40 embryos were collected. Scale bar: 25 µm.

Mentions: In order to confirm the specificity of Ci-E(z) morpholino, we tested whether the Ci-E(z) phenotype could be rescued by injection of a synthetic Ci-E(z) mRNA lacking the morpholino target sequence simultaneously with the Ci-E(z) morpholino. We first tested the MO-E(z)/mRNA-E(z) ratio classically used in Ciona intestinalis publications (Christiaen et al., 2009). Fig. 6A shows the comparison between control, E(z) morphant and rescue phenotypes obtained at late-tailbud stage. The control injection of mRNA-E(z) alone has no phenotypic effect (supplementary material Fig. S4). The rescued embryos presented partial restoration of E(z) protein levels as well as H3K27me3 mark labeling. At a morphological level, even though rescued embryos exhibited some disorganization features, most rescued embryos expressed a partially differentiated notochord that could be clearly identified by the coherent alignment of notochord cells at the tail-head junction (Fig. 6A, right rescue panels).Fig. 6.


Enhancer of zeste acts as a major developmental regulator of Ciona intestinalis embryogenesis.

Le Goff E, Martinand-Mari C, Martin M, Feuillard J, Boublik Y, Godefroy N, Mangeat P, Baghdiguian S, Cavalli G - Biol Open (2015)

Ci-E(z) and H3K27me3 detection in rescue experiments. Comparative indirect immunofluorescence analysis between (A) control embryos, Ci-E(z) morphants, rescue embryos and (B) Ci-E(z) morphants ½ and rescue ½ embryos at late tailbud stage of development. A triple labeling was performed using: actin (Phalloïdin in green), DNA (DAPI, in blue) and antibodies against H3K27me3 or Ci-E(z) protein (in grey). On the right of each DAPI/Phalloïdin merge, the corresponding image of Ci-E(z) or H3K27me3 alone is shown (as indicated) with cell contour drawn in grey. Ab, antibodies; MO-Ctrl, control embryo; mRNA-E(z), mRNA-E(z) embryo; MO-E(z), Ci-E(z) morphant; MO-E(z) ½, embryo injected with half of MO-E(z) concentration; Rescue, mRNA-E(z)+MO-E(z) embryo; Rescue ½, embryo injected with half of MO-E(z) concentration+mRNA-E(z). For three independent experiments, between 20 and 40 embryos were collected. Scale bar: 25 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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BIO010835F6: Ci-E(z) and H3K27me3 detection in rescue experiments. Comparative indirect immunofluorescence analysis between (A) control embryos, Ci-E(z) morphants, rescue embryos and (B) Ci-E(z) morphants ½ and rescue ½ embryos at late tailbud stage of development. A triple labeling was performed using: actin (Phalloïdin in green), DNA (DAPI, in blue) and antibodies against H3K27me3 or Ci-E(z) protein (in grey). On the right of each DAPI/Phalloïdin merge, the corresponding image of Ci-E(z) or H3K27me3 alone is shown (as indicated) with cell contour drawn in grey. Ab, antibodies; MO-Ctrl, control embryo; mRNA-E(z), mRNA-E(z) embryo; MO-E(z), Ci-E(z) morphant; MO-E(z) ½, embryo injected with half of MO-E(z) concentration; Rescue, mRNA-E(z)+MO-E(z) embryo; Rescue ½, embryo injected with half of MO-E(z) concentration+mRNA-E(z). For three independent experiments, between 20 and 40 embryos were collected. Scale bar: 25 µm.
Mentions: In order to confirm the specificity of Ci-E(z) morpholino, we tested whether the Ci-E(z) phenotype could be rescued by injection of a synthetic Ci-E(z) mRNA lacking the morpholino target sequence simultaneously with the Ci-E(z) morpholino. We first tested the MO-E(z)/mRNA-E(z) ratio classically used in Ciona intestinalis publications (Christiaen et al., 2009). Fig. 6A shows the comparison between control, E(z) morphant and rescue phenotypes obtained at late-tailbud stage. The control injection of mRNA-E(z) alone has no phenotypic effect (supplementary material Fig. S4). The rescued embryos presented partial restoration of E(z) protein levels as well as H3K27me3 mark labeling. At a morphological level, even though rescued embryos exhibited some disorganization features, most rescued embryos expressed a partially differentiated notochord that could be clearly identified by the coherent alignment of notochord cells at the tail-head junction (Fig. 6A, right rescue panels).Fig. 6.

Bottom Line: These major phenotypic defects are specifically rescued by injection of a morpholino-resistant Ci-E(z) mRNA, which restores expression of Ci-E(z) protein and re-deposition of the H3K27me3 mark.As observed by qPCR analyses, Ci-E(z) invalidation leads to the early derepression of tissue-specific developmental genes, whereas late-acting developmental genes are generally down-regulated.Altogether, our results suggest that Ci-E(z) plays a major role during embryonic development in Ciona intestinalis by silencing early-acting developmental genes in a Hox-independent manner.

View Article: PubMed Central - PubMed

Affiliation: Université Montpellier, Place Eugène Bataillon, Montpellier 34095, Cedex 5, France Institut des Sciences de l'Evolution (ISEM), UMR5554, CNRS, Montpellier 34095, France.

No MeSH data available.


Related in: MedlinePlus