Involvement of Transcription Factor NR2F2 in Human Trophoblast Differentiation

differentiation of human villous cytotrophoblast (CTB) cells to a syncytiotrophoblast (STB) phenotype, mRNA levels for the nuclear hormone receptor NR2F2 (ARP-1, COUP-TFII) increase rapidly, reaching a peak at day 1 of differentiation that is 8.8-fold greater than that in undifferentiated CTB cells. To examine whether NR2F2 is involved in the regulation of villous CTB cell differentiation, studies were performed to determine whether NR2F2 regulates the expression of TFAP2A (AP-2α), a transcription factor that is critical for the terminal differentiation of these cells to a STB phenotype.Overexpression of NR2F2 in primary cultures of human CTB cells and JEG-3 human choriocarcinoma cells induced dose-dependent increases in TFAP2A promoter activity. Conversely, siRNA mediated silencing of the NR2F2 gene in villous CTB undergoing spontaneous differentiation blocked the induction of the mRNAs for TFAP2A and several STB cell specific marker genes, including human placental lactogen (hPL), pregnancy specific glycoprotein 1 (PSG1) and corticotropin releasing hormone (CRH) by 51–59%. The induction of TFAP2A promoter activity by NR2F2 was potentiated by the nuclear hormone receptors retinoic acid receptor alpha (RARA) and retinoid X receptor alpha (RXRA).Taken together, these results strongly suggest that NR2F2 is involved in villous CTB cell differentiation and that NR2F2 acts, at least in part, by directly activating TFAP2A gene expression and by potentiating the transactivation of TFAP2A by RARA and RXRA

Use of SMS texts for facilitating access to online alcohol interventions: a feasibility study

A41 Use of SMS texts for facilitating access to online alcohol interventions: a feasibility study In: Addiction Science & Clinical Practice 2017, 12(Suppl 1): A4

Proceedings of the 13th annual conference of INEBRIA

CITATION: Watson, R., et al. 2016. Proceedings of the 13th annual conference of INEBRIA. Addiction Science & Clinical Practice, 11:13, doi:10.1186/s13722-016-0062-9.The original publication is available at https://ascpjournal.biomedcentral.comENGLISH SUMMARY : Meeting abstracts.https://ascpjournal.biomedcentral.com/articles/10.1186/s13722-016-0062-9Publisher's versio

Time course of TFAP2A and NR2F2 mRNA levels during villous CTB differentiation.

<p>An enriched fraction of enzymatically dispersed villous CTB cells were cultured <i>in vitro</i> for five days as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009417#s4" target="_blank">Methods</a>. TFAP2A and NR2F2 mRNA levels were determined by real-time PCR at the end of each day. The amounts of TFAP2A and NR2F2 mRNAs in each culture well were normalized to the amount of GAPDH mRNA in the same sample. Each point represents the mean ± SEM of triplicate observations from 3 different placenta cell preparations (n = 3 wells/placenta culture).</p

The effect of RXRA on NR2F2-induced TFAP2A promoter activity.

<p>JEG-3 cells were co-transfected with pGL3β-TFAP2A-Luc and pMT2-NR2F2 in the presence and absence of pRSV-RXRA. An equivalent amount of the empty pRSV plasmid was co-transfected into the cells that were not co-transfected with pRSV-RXRA. Each bar represents the mean of triplicate wells; and the brackets enclose 1 SEM. NS = not significant; * = p<0.05; *** = p<0.001. Similar results were obtained in two other experiments.</p