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Histone acetylation is involved in TCDD‑induced cleft palate formation in fetal mice.

Yuan X, Qiu L, Pu Y, Liu C, Zhang X, Wang C, Pu W, Fu Y - Mol Med Rep (2016)

Bottom Line: On GD 13.5 and GD 14.5, TGF‑β3 mRNA expression, HAT activity and acetylated H3 levels were significantly increased in the TCDD group compared with the control.Methylated bands were not observed in the TCDD or control groups.In conclusion, at the critical period of palate fusion (GD 13.5‑14.5), TCDD significantly increased TGF‑β3 gene expression, HAT activity and H3 acetylation.

View Article: PubMed Central - PubMed

Affiliation: Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing 400014, P.R. China.

ABSTRACT
The aim of the present was to evaluate the effects of DNA methylation and histone acetylation on 2,3,7,8‑tetrachlorodibenzo‑p‑dioxin (TCDD)‑induced cleft palate in fetal mice. Pregnant mice (n=10) were randomly divided into two groups: i) TCDD group, mice were treated with 28 µg/kg TCDD on gestation day (GD) 10 by oral gavage; ii) control group, mice were treated with an equal volume of corn oil. On GD 16.5, the fetal mice were evaluated for the presence of a cleft palate. An additional 36 pregnant mice were divided into the control and TCDD groups, and palate samples were collected on GD 13.5, GD 14.5 and GD 15.5, respectively. Transforming growth factor‑β3 (TGF‑β3) mRNA expression, TGF‑β3 promoter methylation, histone acetyltransferase (HAT) activity and histone H3 (H3) acetylation in the palates were evaluated in the two groups. The incidence of a cleft palate in the TCDD group was 93.55%, and no cases of cleft palate were identified in the control group. On GD 13.5 and GD 14.5, TGF‑β3 mRNA expression, HAT activity and acetylated H3 levels were significantly increased in the TCDD group compared with the control. Methylated bands were not observed in the TCDD or control groups. In conclusion, at the critical period of palate fusion (GD 13.5‑14.5), TCDD significantly increased TGF‑β3 gene expression, HAT activity and H3 acetylation. Therefore, histone acetylation may be involved in TCDD‑induced cleft palate formation in fetal mice.

No MeSH data available.


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Transforming growth factor-β3 CpG island locations.
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f1-mmr-14-02-1139: Transforming growth factor-β3 CpG island locations.

Mentions: Methylation of the TGF-β3 gene promoter was examined using a DNA methylation kit (Zymo Research Corporation, Irvine, CA, USA). The TGF-β3 gene promoter sequences were obtained from the UCSC Genome Browser Home and inputted into MethPrimer software (15). The location of the CpG island in the TGF-β3 gene promoter is shown in Fig. 1, and the primers for methylation analysis were synthesized by Shanghai Biological Engineering Co., Ltd. (Shanghai, China) and are presented in Table I. Total DNA was extracted from the frozen palates using a Cell/Tissue DNA Extraction kit (BioTeke Corporation). PCR was performed in a reaction volume of 25 µl containing 2 µl cDNA, 12.5 µl Zymo Taq PreMix (Zymo Research Corporation), 1 µl of forward and reverse primers (Beijing SBS Genetech Co., Ltd.), and 8.5 µl ddH2O. The PCR parameters were as follows: Denaturation at 95°C for 10 min; 35 cycles of 95°C for 30 sec, 56°C for 40 sec, and 72°C for 60 sec; and a final extension at 72°C for 7 min. Equal quantities (5 µl) of PCR product from each group were analyzed on a 1.5% agarose gel containing ethidium bromide (Shanghai Chemical Reagent), and the target bands were analyzed densitometrically on a Vistra Fluor Imager SI (Molecular Dynamics Inc., Albuquerque, NM, USA).


Histone acetylation is involved in TCDD‑induced cleft palate formation in fetal mice.

Yuan X, Qiu L, Pu Y, Liu C, Zhang X, Wang C, Pu W, Fu Y - Mol Med Rep (2016)

Transforming growth factor-β3 CpG island locations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940082&req=5

f1-mmr-14-02-1139: Transforming growth factor-β3 CpG island locations.
Mentions: Methylation of the TGF-β3 gene promoter was examined using a DNA methylation kit (Zymo Research Corporation, Irvine, CA, USA). The TGF-β3 gene promoter sequences were obtained from the UCSC Genome Browser Home and inputted into MethPrimer software (15). The location of the CpG island in the TGF-β3 gene promoter is shown in Fig. 1, and the primers for methylation analysis were synthesized by Shanghai Biological Engineering Co., Ltd. (Shanghai, China) and are presented in Table I. Total DNA was extracted from the frozen palates using a Cell/Tissue DNA Extraction kit (BioTeke Corporation). PCR was performed in a reaction volume of 25 µl containing 2 µl cDNA, 12.5 µl Zymo Taq PreMix (Zymo Research Corporation), 1 µl of forward and reverse primers (Beijing SBS Genetech Co., Ltd.), and 8.5 µl ddH2O. The PCR parameters were as follows: Denaturation at 95°C for 10 min; 35 cycles of 95°C for 30 sec, 56°C for 40 sec, and 72°C for 60 sec; and a final extension at 72°C for 7 min. Equal quantities (5 µl) of PCR product from each group were analyzed on a 1.5% agarose gel containing ethidium bromide (Shanghai Chemical Reagent), and the target bands were analyzed densitometrically on a Vistra Fluor Imager SI (Molecular Dynamics Inc., Albuquerque, NM, USA).

Bottom Line: On GD 13.5 and GD 14.5, TGF‑β3 mRNA expression, HAT activity and acetylated H3 levels were significantly increased in the TCDD group compared with the control.Methylated bands were not observed in the TCDD or control groups.In conclusion, at the critical period of palate fusion (GD 13.5‑14.5), TCDD significantly increased TGF‑β3 gene expression, HAT activity and H3 acetylation.

View Article: PubMed Central - PubMed

Affiliation: Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing 400014, P.R. China.

ABSTRACT
The aim of the present was to evaluate the effects of DNA methylation and histone acetylation on 2,3,7,8‑tetrachlorodibenzo‑p‑dioxin (TCDD)‑induced cleft palate in fetal mice. Pregnant mice (n=10) were randomly divided into two groups: i) TCDD group, mice were treated with 28 µg/kg TCDD on gestation day (GD) 10 by oral gavage; ii) control group, mice were treated with an equal volume of corn oil. On GD 16.5, the fetal mice were evaluated for the presence of a cleft palate. An additional 36 pregnant mice were divided into the control and TCDD groups, and palate samples were collected on GD 13.5, GD 14.5 and GD 15.5, respectively. Transforming growth factor‑β3 (TGF‑β3) mRNA expression, TGF‑β3 promoter methylation, histone acetyltransferase (HAT) activity and histone H3 (H3) acetylation in the palates were evaluated in the two groups. The incidence of a cleft palate in the TCDD group was 93.55%, and no cases of cleft palate were identified in the control group. On GD 13.5 and GD 14.5, TGF‑β3 mRNA expression, HAT activity and acetylated H3 levels were significantly increased in the TCDD group compared with the control. Methylated bands were not observed in the TCDD or control groups. In conclusion, at the critical period of palate fusion (GD 13.5‑14.5), TCDD significantly increased TGF‑β3 gene expression, HAT activity and H3 acetylation. Therefore, histone acetylation may be involved in TCDD‑induced cleft palate formation in fetal mice.

No MeSH data available.


Related in: MedlinePlus