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Glucocorticoid receptor is involved in the breed-dependent transcriptional regulation of 3β-hydroxysteroid dehydrogenase in the liver of preweaning piglets.

Li X, Jia Y, Li R, Sun Z, Li X, Sui S, Zhao R - BMC Vet. Res. (2015)

Bottom Line: The hepatic expression of 3β-HSD mRNA was significantly lower (P < 0.01) in EHL piglets compared to that in LW piglets.The nucleoprotein contents of AR (P < 0.05), GR (P < 0.01) and phospho-Ser(211)GR (P < 0.01) were significantly higher in the liver of EHL piglets.These results indicate that GR binding to 3β-HSD promoter is involved in the breed-dependent 3β-HSD expression in the liver of piglets.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Physiology and Biochemistry, Nanjing Agricultural University, Nanjing, 210095, P. R. China. lixianlovelv@163.com.

ABSTRACT

Background: Hepatic 3β-hydroxysteroid dehydrogenase (3β-HSD) plays an important role in steroid inactivation and catabolism. Serum concentrations of steroid hormones differ significantly between breeds in pigs, however the molecular mechanism regulating hepatic 3β-HSD expression in different breeds of pigs is poorly understood. In the present study, we used preweaning purebred male Large White (LW) and Erhualian (EHL) piglets as model to investigate the breed difference in the expression and regulation of 3β-HSD gene in porcine liver.

Results: The hepatic expression of 3β-HSD mRNA was significantly lower (P < 0.01) in EHL piglets compared to that in LW piglets. Significant breed differences were detected for the hepatic expression of transcription factors such as androgen receptor (AR), glucocorticoid receptor (GR), and CCAAT/enhancer binding protein β (C/EBPβ). The nucleoprotein contents of AR (P < 0.05), GR (P < 0.01) and phospho-Ser(211)GR (P < 0.01) were significantly higher in the liver of EHL piglets. Chromatin immunoprecipitation (ChIP) assay demonstrated significantly lower binding of GR, but not AR or C/EBPβ, to 3β-HSD gene promoter in EHL piglets (P < 0.05). GR was not detected to interact with C/EBPβ or AR in the co-immunoprecipitation analysis.

Conclusions: These results indicate that GR binding to 3β-HSD promoter is involved in the breed-dependent 3β-HSD expression in the liver of piglets.

No MeSH data available.


ChIP analysis for GR, AR and C/EBPβ binding on hepatic 3β-HSD promoter. (a) Schematic structure of porcine 3β-HSD promoter. GRE, ARE and C/EBPβ binding sites and the PCR fragments used to detect GR, AR and C/EBPβ binding are indicated; TSS: transcription start site. (b) GR binding on hepatic 3β-HSD promoter. (c) AR binding on hepatic 3β-HSD promoter. (d) C/EBPβ binding on hepatic 3β-HSD promoter. Values are means ± SEM, n = 5 for LW or 6 for EHL. * P < 0.05, compared with LW
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Fig3: ChIP analysis for GR, AR and C/EBPβ binding on hepatic 3β-HSD promoter. (a) Schematic structure of porcine 3β-HSD promoter. GRE, ARE and C/EBPβ binding sites and the PCR fragments used to detect GR, AR and C/EBPβ binding are indicated; TSS: transcription start site. (b) GR binding on hepatic 3β-HSD promoter. (c) AR binding on hepatic 3β-HSD promoter. (d) C/EBPβ binding on hepatic 3β-HSD promoter. Values are means ± SEM, n = 5 for LW or 6 for EHL. * P < 0.05, compared with LW

Mentions: Three putative GREs, 3 AREs and 4 C/EBPβ binding sites were predicted on pig 3β-HSD gene promoter (Fig. 3a). ChIP assay revealed significantly lower binding of GR to the first and the third predicted GREs on 3β-HSD promoter in the liver of EHL piglets (P < 0.05, Fig. 3b). No significant breed difference was observed for the binding of AR or C/EBPβ to 3β-HSD promoter in the liver of preweaning piglets (Fig. 3c and d).Fig. 3


Glucocorticoid receptor is involved in the breed-dependent transcriptional regulation of 3β-hydroxysteroid dehydrogenase in the liver of preweaning piglets.

Li X, Jia Y, Li R, Sun Z, Li X, Sui S, Zhao R - BMC Vet. Res. (2015)

ChIP analysis for GR, AR and C/EBPβ binding on hepatic 3β-HSD promoter. (a) Schematic structure of porcine 3β-HSD promoter. GRE, ARE and C/EBPβ binding sites and the PCR fragments used to detect GR, AR and C/EBPβ binding are indicated; TSS: transcription start site. (b) GR binding on hepatic 3β-HSD promoter. (c) AR binding on hepatic 3β-HSD promoter. (d) C/EBPβ binding on hepatic 3β-HSD promoter. Values are means ± SEM, n = 5 for LW or 6 for EHL. * P < 0.05, compared with LW
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig3: ChIP analysis for GR, AR and C/EBPβ binding on hepatic 3β-HSD promoter. (a) Schematic structure of porcine 3β-HSD promoter. GRE, ARE and C/EBPβ binding sites and the PCR fragments used to detect GR, AR and C/EBPβ binding are indicated; TSS: transcription start site. (b) GR binding on hepatic 3β-HSD promoter. (c) AR binding on hepatic 3β-HSD promoter. (d) C/EBPβ binding on hepatic 3β-HSD promoter. Values are means ± SEM, n = 5 for LW or 6 for EHL. * P < 0.05, compared with LW
Mentions: Three putative GREs, 3 AREs and 4 C/EBPβ binding sites were predicted on pig 3β-HSD gene promoter (Fig. 3a). ChIP assay revealed significantly lower binding of GR to the first and the third predicted GREs on 3β-HSD promoter in the liver of EHL piglets (P < 0.05, Fig. 3b). No significant breed difference was observed for the binding of AR or C/EBPβ to 3β-HSD promoter in the liver of preweaning piglets (Fig. 3c and d).Fig. 3

Bottom Line: The hepatic expression of 3β-HSD mRNA was significantly lower (P < 0.01) in EHL piglets compared to that in LW piglets.The nucleoprotein contents of AR (P < 0.05), GR (P < 0.01) and phospho-Ser(211)GR (P < 0.01) were significantly higher in the liver of EHL piglets.These results indicate that GR binding to 3β-HSD promoter is involved in the breed-dependent 3β-HSD expression in the liver of piglets.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Physiology and Biochemistry, Nanjing Agricultural University, Nanjing, 210095, P. R. China. lixianlovelv@163.com.

ABSTRACT

Background: Hepatic 3β-hydroxysteroid dehydrogenase (3β-HSD) plays an important role in steroid inactivation and catabolism. Serum concentrations of steroid hormones differ significantly between breeds in pigs, however the molecular mechanism regulating hepatic 3β-HSD expression in different breeds of pigs is poorly understood. In the present study, we used preweaning purebred male Large White (LW) and Erhualian (EHL) piglets as model to investigate the breed difference in the expression and regulation of 3β-HSD gene in porcine liver.

Results: The hepatic expression of 3β-HSD mRNA was significantly lower (P < 0.01) in EHL piglets compared to that in LW piglets. Significant breed differences were detected for the hepatic expression of transcription factors such as androgen receptor (AR), glucocorticoid receptor (GR), and CCAAT/enhancer binding protein β (C/EBPβ). The nucleoprotein contents of AR (P < 0.05), GR (P < 0.01) and phospho-Ser(211)GR (P < 0.01) were significantly higher in the liver of EHL piglets. Chromatin immunoprecipitation (ChIP) assay demonstrated significantly lower binding of GR, but not AR or C/EBPβ, to 3β-HSD gene promoter in EHL piglets (P < 0.05). GR was not detected to interact with C/EBPβ or AR in the co-immunoprecipitation analysis.

Conclusions: These results indicate that GR binding to 3β-HSD promoter is involved in the breed-dependent 3β-HSD expression in the liver of piglets.

No MeSH data available.