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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) Western blot with the anti–ING2 antibody showed only one distinct band at the predicted molecular weight of ING2 protein (33 kD), for both the mouse ovary and 4-cell embryo. (B) Immunofluorescence staining of ING2 and nuclear DNA in preimplantation embryos with specific anti-ING2 antibody (green, a′–f′) and propidium iodide (red, a′′–f′′) respectively. zygote, approximately 21 h after human chorionic gonadotropin (HCG) injection (a′, a′′); 2-cell embryo, approximately 45 h after HCG injection (b′, b′′); 4- to 8-cell embryo, approximately 58 h after HCG injection (c′, c′′, d′, d′′); morula, approximately 78 h after HCG injection (e,′ e′′) and blastocyst, approximately 96 h after HCG injection (f′, f′′). Scale bar: 2 μm. (C) Unstained 2-cell embryo was utilized as the negative control. Scale bar: 1 μm.
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fig002: (A) Western blot with the anti–ING2 antibody showed only one distinct band at the predicted molecular weight of ING2 protein (33 kD), for both the mouse ovary and 4-cell embryo. (B) Immunofluorescence staining of ING2 and nuclear DNA in preimplantation embryos with specific anti-ING2 antibody (green, a′–f′) and propidium iodide (red, a′′–f′′) respectively. zygote, approximately 21 h after human chorionic gonadotropin (HCG) injection (a′, a′′); 2-cell embryo, approximately 45 h after HCG injection (b′, b′′); 4- to 8-cell embryo, approximately 58 h after HCG injection (c′, c′′, d′, d′′); morula, approximately 78 h after HCG injection (e,′ e′′) and blastocyst, approximately 96 h after HCG injection (f′, f′′). Scale bar: 2 μm. (C) Unstained 2-cell embryo was utilized as the negative control. Scale bar: 1 μm.

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) Western blot with the anti–ING2 antibody showed only one distinct band at the predicted molecular weight of ING2 protein (33 kD), for both the mouse ovary and 4-cell embryo. (B) Immunofluorescence staining of ING2 and nuclear DNA in preimplantation embryos with specific anti-ING2 antibody (green, a′–f′) and propidium iodide (red, a′′–f′′) respectively. zygote, approximately 21 h after human chorionic gonadotropin (HCG) injection (a′, a′′); 2-cell embryo, approximately 45 h after HCG injection (b′, b′′); 4- to 8-cell embryo, approximately 58 h after HCG injection (c′, c′′, d′, d′′); morula, approximately 78 h after HCG injection (e,′ e′′) and blastocyst, approximately 96 h after HCG injection (f′, f′′). Scale bar: 2 μm. (C) Unstained 2-cell embryo was utilized as the negative control. Scale bar: 1 μm.
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Related In: Results  -  Collection

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fig002: (A) Western blot with the anti–ING2 antibody showed only one distinct band at the predicted molecular weight of ING2 protein (33 kD), for both the mouse ovary and 4-cell embryo. (B) Immunofluorescence staining of ING2 and nuclear DNA in preimplantation embryos with specific anti-ING2 antibody (green, a′–f′) and propidium iodide (red, a′′–f′′) respectively. zygote, approximately 21 h after human chorionic gonadotropin (HCG) injection (a′, a′′); 2-cell embryo, approximately 45 h after HCG injection (b′, b′′); 4- to 8-cell embryo, approximately 58 h after HCG injection (c′, c′′, d′, d′′); morula, approximately 78 h after HCG injection (e,′ e′′) and blastocyst, approximately 96 h after HCG injection (f′, f′′). Scale bar: 2 μm. (C) Unstained 2-cell embryo was utilized as the negative control. Scale bar: 1 μm.
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