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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

Phenotypes obtained in rescue experiments at the 4-cell stage. (A) Comparative indirect immunofluorescence analysis between control embryos, Ci-E(z) morphants, Ci-E(z) morphants injected with half of the MO-E(z) concentration, rescue embryos and rescue ½ embryos at 4-cell stage with a double labeling: actin (Phalloïdin, green) and DNA (DAPI, blue). MO-Ctrl, control embryo; MO-E(z), Ci-E(z) morphant; MO-E(z) ½, morphant 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 each point, between 20 and 40 embryos were collected. Scale bar: 25 µm. (B) Corresponding percentage of embryos categorized as normal (N) or types 1–2. Type 1: Blastomeres are distinguishable from each other, embryo symmetry is not preserved. Type 2: Blastomeres are not properly separated, embryo symmetry is lost.
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BIO010835F7: Phenotypes obtained in rescue experiments at the 4-cell stage. (A) Comparative indirect immunofluorescence analysis between control embryos, Ci-E(z) morphants, Ci-E(z) morphants injected with half of the MO-E(z) concentration, rescue embryos and rescue ½ embryos at 4-cell stage with a double labeling: actin (Phalloïdin, green) and DNA (DAPI, blue). MO-Ctrl, control embryo; MO-E(z), Ci-E(z) morphant; MO-E(z) ½, morphant 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 each point, between 20 and 40 embryos were collected. Scale bar: 25 µm. (B) Corresponding percentage of embryos categorized as normal (N) or types 1–2. Type 1: Blastomeres are distinguishable from each other, embryo symmetry is not preserved. Type 2: Blastomeres are not properly separated, embryo symmetry is lost.

Mentions: The results being striking at the larval stage, is the rescue active as soon as at the 4-cell stage? To answer this question, we performed rescue assays at this earlier stage when the E(z) inhibition is already effective (Figs 2 and 7). While both concentrations of MO-E(z) caused cytokinesis defects, co-injections of MO-E(z) with mRNA-E(z) restored a control phenotype with, notably, the rescue of the characteristic 4-cell symmetrical organization (Fig. 7, rescue and rescue ½ panels). These data show that Ci-E(z) mRNA injection does partially rescue both early and late effects of its impairment by morpholino, further substantiating the early and crucial role of this protein in Ciona intestinalis embryogenesis.Fig. 7.


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)

Phenotypes obtained in rescue experiments at the 4-cell stage. (A) Comparative indirect immunofluorescence analysis between control embryos, Ci-E(z) morphants, Ci-E(z) morphants injected with half of the MO-E(z) concentration, rescue embryos and rescue ½ embryos at 4-cell stage with a double labeling: actin (Phalloïdin, green) and DNA (DAPI, blue). MO-Ctrl, control embryo; MO-E(z), Ci-E(z) morphant; MO-E(z) ½, morphant 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 each point, between 20 and 40 embryos were collected. Scale bar: 25 µm. (B) Corresponding percentage of embryos categorized as normal (N) or types 1–2. Type 1: Blastomeres are distinguishable from each other, embryo symmetry is not preserved. Type 2: Blastomeres are not properly separated, embryo symmetry is lost.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4582116&req=5

BIO010835F7: Phenotypes obtained in rescue experiments at the 4-cell stage. (A) Comparative indirect immunofluorescence analysis between control embryos, Ci-E(z) morphants, Ci-E(z) morphants injected with half of the MO-E(z) concentration, rescue embryos and rescue ½ embryos at 4-cell stage with a double labeling: actin (Phalloïdin, green) and DNA (DAPI, blue). MO-Ctrl, control embryo; MO-E(z), Ci-E(z) morphant; MO-E(z) ½, morphant 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 each point, between 20 and 40 embryos were collected. Scale bar: 25 µm. (B) Corresponding percentage of embryos categorized as normal (N) or types 1–2. Type 1: Blastomeres are distinguishable from each other, embryo symmetry is not preserved. Type 2: Blastomeres are not properly separated, embryo symmetry is lost.
Mentions: The results being striking at the larval stage, is the rescue active as soon as at the 4-cell stage? To answer this question, we performed rescue assays at this earlier stage when the E(z) inhibition is already effective (Figs 2 and 7). While both concentrations of MO-E(z) caused cytokinesis defects, co-injections of MO-E(z) with mRNA-E(z) restored a control phenotype with, notably, the rescue of the characteristic 4-cell symmetrical organization (Fig. 7, rescue and rescue ½ panels). These data show that Ci-E(z) mRNA injection does partially rescue both early and late effects of its impairment by morpholino, further substantiating the early and crucial role of this protein in Ciona intestinalis embryogenesis.Fig. 7.

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