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The Anti-Adipogenic Potential of COUP-TFII Is Mediated by Downregulation of the Notch Target Gene Hey1.

Scroyen I, Bauters D, Vranckx C, Lijnen HR - PLoS ONE (2015)

Bottom Line: It has not conclusively been established, however, whether its role is pro- or anti-adipogenic.Gene silencing of Coup-tfII in 3T3-F442A preadipocytes resulted in enhanced differentiation into mature adipocytes.A functional role of Hey1 was confirmed by gene silencing in 3T3-F442A preadipocytes, resulting in impaired differentiation.

View Article: PubMed Central - PubMed

Affiliation: KU Leuven, University of Leuven, Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, B-3000, Leuven, Belgium.

ABSTRACT

Background: Chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) belongs to the steroid/thyroid hormone receptor superfamily and may contribute to the pathogenesis of obesity. It has not conclusively been established, however, whether its role is pro- or anti-adipogenic.

Methods and results: Gene silencing of Coup-tfII in 3T3-F442A preadipocytes resulted in enhanced differentiation into mature adipocytes. This was associated with upregulation of the Notch signaling target gene Hey1. A functional role of Hey1 was confirmed by gene silencing in 3T3-F442A preadipocytes, resulting in impaired differentiation. In vivo, de novo fat pad formation in NUDE mice was significantly stimulated following injection of preadipocytes with Coup-tfII gene silencing, but impaired with Hey1 gene silencing. Moreover, expression of Coup-tfII was lower and that of Hey1 higher in isolated adipocytes of obese as compared to lean adipose tissue.

Conclusions: These in vitro and in vivo data support an anti-adipogenic role of COUP-TFII via downregulating the Notch signaling target gene Hey1.

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Effect of Hey1 gene silencing on in vitro differentiation of 3T3-F442A preadipocytes.(A) Time course of Hey1 expression during differentiation without (⚫, black circles) or with (○, open circles) knockdown (kd). (B-C) Oil Red O staining and quantification (OD at 490 nm) at day 12 of differentiation. (D-J) Time course of expression of aP2 (D), PPAR-γ (E), GPDH (F), GLUT4 (G) C/EBPα (H), C/EBPβ (I) and C/EBPδ (J) during differentiation without (⚫, black circles) or with (○, open circles) knockdown (kd). Data are means ± SEM of 3 independent experiments; *** p < 0.001 **** p < 0.0001. The scale bar in panel C corresponds to 100 μm
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pone.0145608.g002: Effect of Hey1 gene silencing on in vitro differentiation of 3T3-F442A preadipocytes.(A) Time course of Hey1 expression during differentiation without (⚫, black circles) or with (○, open circles) knockdown (kd). (B-C) Oil Red O staining and quantification (OD at 490 nm) at day 12 of differentiation. (D-J) Time course of expression of aP2 (D), PPAR-γ (E), GPDH (F), GLUT4 (G) C/EBPα (H), C/EBPβ (I) and C/EBPδ (J) during differentiation without (⚫, black circles) or with (○, open circles) knockdown (kd). Data are means ± SEM of 3 independent experiments; *** p < 0.001 **** p < 0.0001. The scale bar in panel C corresponds to 100 μm

Mentions: During differentiation of control preadipocytes, Hey1 mRNA expression gradually increased as a function of time, whereas Hey1 gene silencing (Fig 2A) was stable during the 12day-differentiation period. Oil Red O analysis of the intra-cytoplasmatic lipid content of differentiated preadipocytes confirmed impaired differentiation upon Hey1 silencing (Fig 2B and 2C). Monitoring of adipogenic markers including aP2, PPAR-γ, GPDH, GLUT4 and C/EBPα during differentiation confirmed lower expression in Hey1 knock down cells as compared to control cells (Fig 2D–2H). No differences in the expression of C/EBPβ, C/EBPδ (Fig 2I and 2J) Notch1, Notch2 or the target gene Hes1 were detected (S2B–S2D Fig).


The Anti-Adipogenic Potential of COUP-TFII Is Mediated by Downregulation of the Notch Target Gene Hey1.

Scroyen I, Bauters D, Vranckx C, Lijnen HR - PLoS ONE (2015)

Effect of Hey1 gene silencing on in vitro differentiation of 3T3-F442A preadipocytes.(A) Time course of Hey1 expression during differentiation without (⚫, black circles) or with (○, open circles) knockdown (kd). (B-C) Oil Red O staining and quantification (OD at 490 nm) at day 12 of differentiation. (D-J) Time course of expression of aP2 (D), PPAR-γ (E), GPDH (F), GLUT4 (G) C/EBPα (H), C/EBPβ (I) and C/EBPδ (J) during differentiation without (⚫, black circles) or with (○, open circles) knockdown (kd). Data are means ± SEM of 3 independent experiments; *** p < 0.001 **** p < 0.0001. The scale bar in panel C corresponds to 100 μm
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4697848&req=5

pone.0145608.g002: Effect of Hey1 gene silencing on in vitro differentiation of 3T3-F442A preadipocytes.(A) Time course of Hey1 expression during differentiation without (⚫, black circles) or with (○, open circles) knockdown (kd). (B-C) Oil Red O staining and quantification (OD at 490 nm) at day 12 of differentiation. (D-J) Time course of expression of aP2 (D), PPAR-γ (E), GPDH (F), GLUT4 (G) C/EBPα (H), C/EBPβ (I) and C/EBPδ (J) during differentiation without (⚫, black circles) or with (○, open circles) knockdown (kd). Data are means ± SEM of 3 independent experiments; *** p < 0.001 **** p < 0.0001. The scale bar in panel C corresponds to 100 μm
Mentions: During differentiation of control preadipocytes, Hey1 mRNA expression gradually increased as a function of time, whereas Hey1 gene silencing (Fig 2A) was stable during the 12day-differentiation period. Oil Red O analysis of the intra-cytoplasmatic lipid content of differentiated preadipocytes confirmed impaired differentiation upon Hey1 silencing (Fig 2B and 2C). Monitoring of adipogenic markers including aP2, PPAR-γ, GPDH, GLUT4 and C/EBPα during differentiation confirmed lower expression in Hey1 knock down cells as compared to control cells (Fig 2D–2H). No differences in the expression of C/EBPβ, C/EBPδ (Fig 2I and 2J) Notch1, Notch2 or the target gene Hes1 were detected (S2B–S2D Fig).

Bottom Line: It has not conclusively been established, however, whether its role is pro- or anti-adipogenic.Gene silencing of Coup-tfII in 3T3-F442A preadipocytes resulted in enhanced differentiation into mature adipocytes.A functional role of Hey1 was confirmed by gene silencing in 3T3-F442A preadipocytes, resulting in impaired differentiation.

View Article: PubMed Central - PubMed

Affiliation: KU Leuven, University of Leuven, Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, B-3000, Leuven, Belgium.

ABSTRACT

Background: Chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) belongs to the steroid/thyroid hormone receptor superfamily and may contribute to the pathogenesis of obesity. It has not conclusively been established, however, whether its role is pro- or anti-adipogenic.

Methods and results: Gene silencing of Coup-tfII in 3T3-F442A preadipocytes resulted in enhanced differentiation into mature adipocytes. This was associated with upregulation of the Notch signaling target gene Hey1. A functional role of Hey1 was confirmed by gene silencing in 3T3-F442A preadipocytes, resulting in impaired differentiation. In vivo, de novo fat pad formation in NUDE mice was significantly stimulated following injection of preadipocytes with Coup-tfII gene silencing, but impaired with Hey1 gene silencing. Moreover, expression of Coup-tfII was lower and that of Hey1 higher in isolated adipocytes of obese as compared to lean adipose tissue.

Conclusions: These in vitro and in vivo data support an anti-adipogenic role of COUP-TFII via downregulating the Notch signaling target gene Hey1.

Show MeSH
Related in: MedlinePlus