Limits...
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

mRNA expression of Ci-E(z) in wild-type embryos and Ci-E(z) morphants. Time course of Ci-E(z) mRNA expression between 1-cell stage to hatching stage in Ci-E(z) morphants (light grey) and wild-type embryos (dark grey). Histograms are the mean of four independent micro-injection experiments; data were normalized to respective S26 mRNA expression values. The “relative mRNA quantity” was expressed with a value of 1 set as the amount of mRNA at 1-cell stage. No significant difference in mRNA levels was observed between wild-type and morphants at any stage. Error bars correspond to the standard deviation (s.d.) from four independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4582116&req=5

BIO010835F1: mRNA expression of Ci-E(z) in wild-type embryos and Ci-E(z) morphants. Time course of Ci-E(z) mRNA expression between 1-cell stage to hatching stage in Ci-E(z) morphants (light grey) and wild-type embryos (dark grey). Histograms are the mean of four independent micro-injection experiments; data were normalized to respective S26 mRNA expression values. The “relative mRNA quantity” was expressed with a value of 1 set as the amount of mRNA at 1-cell stage. No significant difference in mRNA levels was observed between wild-type and morphants at any stage. Error bars correspond to the standard deviation (s.d.) from four independent experiments.

Mentions: Ci-E(z) gene is maternally expressed and its relative mRNA content is maximal at the 64-cell stage and decreases gradually over time (Fig. 1). In order to repress Ci-E(z) function, Ciona intestinalis eggs were injected with either Ci-E(z) or control morpholinos. Two Ci-E(z) morpholinos were designed with the aim to target the AUG codon and generate untranslatable mRNAs. Both morpholinos induced the same phenotype (data not shown), so only one (#1, see Materials and Methods) of them was used in further experiments. Following morpholino injection we verified, by qPCR, the level of mRNA expression of Ci-E(z) (Fig. 1). No significant difference between control and morphant embryos was observed, consistent with the fact that injection of Ci-E(z) morpholino should only induce a defective expression of the protein. In order to detect Ci-E(z) protein, we raised a specific antibody from a recombinant N-terminal fragment of Ci-E(z).Fig. 1.


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)

mRNA expression of Ci-E(z) in wild-type embryos and Ci-E(z) morphants. Time course of Ci-E(z) mRNA expression between 1-cell stage to hatching stage in Ci-E(z) morphants (light grey) and wild-type embryos (dark grey). Histograms are the mean of four independent micro-injection experiments; data were normalized to respective S26 mRNA expression values. The “relative mRNA quantity” was expressed with a value of 1 set as the amount of mRNA at 1-cell stage. No significant difference in mRNA levels was observed between wild-type and morphants at any stage. Error bars correspond to the standard deviation (s.d.) from four independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

BIO010835F1: mRNA expression of Ci-E(z) in wild-type embryos and Ci-E(z) morphants. Time course of Ci-E(z) mRNA expression between 1-cell stage to hatching stage in Ci-E(z) morphants (light grey) and wild-type embryos (dark grey). Histograms are the mean of four independent micro-injection experiments; data were normalized to respective S26 mRNA expression values. The “relative mRNA quantity” was expressed with a value of 1 set as the amount of mRNA at 1-cell stage. No significant difference in mRNA levels was observed between wild-type and morphants at any stage. Error bars correspond to the standard deviation (s.d.) from four independent experiments.
Mentions: Ci-E(z) gene is maternally expressed and its relative mRNA content is maximal at the 64-cell stage and decreases gradually over time (Fig. 1). In order to repress Ci-E(z) function, Ciona intestinalis eggs were injected with either Ci-E(z) or control morpholinos. Two Ci-E(z) morpholinos were designed with the aim to target the AUG codon and generate untranslatable mRNAs. Both morpholinos induced the same phenotype (data not shown), so only one (#1, see Materials and Methods) of them was used in further experiments. Following morpholino injection we verified, by qPCR, the level of mRNA expression of Ci-E(z) (Fig. 1). No significant difference between control and morphant embryos was observed, consistent with the fact that injection of Ci-E(z) morpholino should only induce a defective expression of the protein. In order to detect Ci-E(z) protein, we raised a specific antibody from a recombinant N-terminal fragment of Ci-E(z).Fig. 1.

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