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ING2 (inhibitor of growth protein-2) plays a crucial role in preimplantation development.

Zhou L, Wang P, Zhang J, Heng BC, Tong GQ - Zygote (2015)

Bottom Line: Embryonic cells microinjected with ING2-specific esiRNA exhibited decreased blastulation rate compared to the negative control.Further investigation of the underlying mechanism indicated that down-regulation of ING2 significantly increased expression of p21, whilst decreasing expression of HDAC1.These results suggest that ING2 may play a crucial role in the process of preimplantation embryo development through chromatin regulation.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Medicine,Department of Reproduction,Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University,Nanjing 210004,PR China.

ABSTRACT
ING2 (inhibitor of growth protein-2) is a member of the ING-gene family and participates in diverse cellular processes involving tumor suppression, DNA repair, cell cycle regulation, and cellular senescence. As a subunit of the Sin3 histone deacetylase complex co-repressor complex, ING2 binds to H3K4me3 to regulate chromatin modification and gene expression. Additionally, ING2 recruits histone methyltransferase (HMT) activity for gene repression, which is independent of the HDAC class I or II pathway. However, the physiological function of ING2 in mouse preimplantation embryo development has not yet been characterized previously. The expression, localization and function of ING2 during preimplantation development were investigated in this study. We showed increasing expression of ING2 within the nucleus from the 4-cell embryo stage onwards; and that down-regulation of ING2 expression by endoribonuclease-prepared small interfering RNA (esiRNA) microinjection results in developmental arrest during the morula to blastocyst transition. Embryonic cells microinjected with ING2-specific esiRNA exhibited decreased blastulation rate compared to the negative control. Further investigation of the underlying mechanism indicated that down-regulation of ING2 significantly increased expression of p21, whilst decreasing expression of HDAC1. These results suggest that ING2 may play a crucial role in the process of preimplantation embryo development through chromatin regulation.

No MeSH data available.


Related in: MedlinePlus

(A) SDS-PAGE gel of RT-PCR amplification of Ing2 mRNA transcripts extracted at different embryonic stages. H3f3a was utilized as the endogenous reference gene. Lanes 1, 2, 3, 4, 5 corresponds to zygote, 2-cell embryo, 4-cell embryo, morula-stage embryo and blastocyst-stage embryo respectively. (B) Real-time PCR analysis of Ing2 mRNA expression patterns at different embryonic stages during mouse preimplantation development. The expression level was calculated from the cycle threshold (Ct) values by the 2−ΔΔCt method. The calibration sample was embryos at the zygote stage. Bar graphs indicate mean ± standard deviation (SD) of three experimental replicates.
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fig001: (A) SDS-PAGE gel of RT-PCR amplification of Ing2 mRNA transcripts extracted at different embryonic stages. H3f3a was utilized as the endogenous reference gene. Lanes 1, 2, 3, 4, 5 corresponds to zygote, 2-cell embryo, 4-cell embryo, morula-stage embryo and blastocyst-stage embryo respectively. (B) Real-time PCR analysis of Ing2 mRNA expression patterns at different embryonic stages during mouse preimplantation development. The expression level was calculated from the cycle threshold (Ct) values by the 2−ΔΔCt method. The calibration sample was embryos at the zygote stage. Bar graphs indicate mean ± standard deviation (SD) of three experimental replicates.

Mentions: We analyzed the expression profile of Ing2 at each stage of mouse preimplantation embryonic development. The RT-PCR data confirmed expression of Ing2 in mouse preimplantation embryos (Fig. 1A), and further analysis revealed variation in expression levels of Ing2 at different embryonic developmental stages (Fig. 1B). Most notably, Ing2 expression rapidly increased from the 2-cell to 4-cell cleavage-stage. This suggests that Ing2 may be a crucial modulator of early embryonic development genes. Western blot analysis utilizing an anti-ING2 antibody revealed an exclusive band at the expected molecular mass of 33 kDa, for both the mouse ovary and 4-cell embryo (Fig. 2A).Figure 1


ING2 (inhibitor of growth protein-2) plays a crucial role in preimplantation development.

Zhou L, Wang P, Zhang J, Heng BC, Tong GQ - Zygote (2015)

(A) SDS-PAGE gel of RT-PCR amplification of Ing2 mRNA transcripts extracted at different embryonic stages. H3f3a was utilized as the endogenous reference gene. Lanes 1, 2, 3, 4, 5 corresponds to zygote, 2-cell embryo, 4-cell embryo, morula-stage embryo and blastocyst-stage embryo respectively. (B) Real-time PCR analysis of Ing2 mRNA expression patterns at different embryonic stages during mouse preimplantation development. The expression level was calculated from the cycle threshold (Ct) values by the 2−ΔΔCt method. The calibration sample was embryos at the zygote stage. Bar graphs indicate mean ± standard deviation (SD) of three experimental replicates.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4697306&req=5

fig001: (A) SDS-PAGE gel of RT-PCR amplification of Ing2 mRNA transcripts extracted at different embryonic stages. H3f3a was utilized as the endogenous reference gene. Lanes 1, 2, 3, 4, 5 corresponds to zygote, 2-cell embryo, 4-cell embryo, morula-stage embryo and blastocyst-stage embryo respectively. (B) Real-time PCR analysis of Ing2 mRNA expression patterns at different embryonic stages during mouse preimplantation development. The expression level was calculated from the cycle threshold (Ct) values by the 2−ΔΔCt method. The calibration sample was embryos at the zygote stage. Bar graphs indicate mean ± standard deviation (SD) of three experimental replicates.
Mentions: We analyzed the expression profile of Ing2 at each stage of mouse preimplantation embryonic development. The RT-PCR data confirmed expression of Ing2 in mouse preimplantation embryos (Fig. 1A), and further analysis revealed variation in expression levels of Ing2 at different embryonic developmental stages (Fig. 1B). Most notably, Ing2 expression rapidly increased from the 2-cell to 4-cell cleavage-stage. This suggests that Ing2 may be a crucial modulator of early embryonic development genes. Western blot analysis utilizing an anti-ING2 antibody revealed an exclusive band at the expected molecular mass of 33 kDa, for both the mouse ovary and 4-cell embryo (Fig. 2A).Figure 1

Bottom Line: Embryonic cells microinjected with ING2-specific esiRNA exhibited decreased blastulation rate compared to the negative control.Further investigation of the underlying mechanism indicated that down-regulation of ING2 significantly increased expression of p21, whilst decreasing expression of HDAC1.These results suggest that ING2 may play a crucial role in the process of preimplantation embryo development through chromatin regulation.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Medicine,Department of Reproduction,Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University,Nanjing 210004,PR China.

ABSTRACT
ING2 (inhibitor of growth protein-2) is a member of the ING-gene family and participates in diverse cellular processes involving tumor suppression, DNA repair, cell cycle regulation, and cellular senescence. As a subunit of the Sin3 histone deacetylase complex co-repressor complex, ING2 binds to H3K4me3 to regulate chromatin modification and gene expression. Additionally, ING2 recruits histone methyltransferase (HMT) activity for gene repression, which is independent of the HDAC class I or II pathway. However, the physiological function of ING2 in mouse preimplantation embryo development has not yet been characterized previously. The expression, localization and function of ING2 during preimplantation development were investigated in this study. We showed increasing expression of ING2 within the nucleus from the 4-cell embryo stage onwards; and that down-regulation of ING2 expression by endoribonuclease-prepared small interfering RNA (esiRNA) microinjection results in developmental arrest during the morula to blastocyst transition. Embryonic cells microinjected with ING2-specific esiRNA exhibited decreased blastulation rate compared to the negative control. Further investigation of the underlying mechanism indicated that down-regulation of ING2 significantly increased expression of p21, whilst decreasing expression of HDAC1. These results suggest that ING2 may play a crucial role in the process of preimplantation embryo development through chromatin regulation.

No MeSH data available.


Related in: MedlinePlus