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Experimental and bioinformatic analysis of cultured Bovine Endometrial Cells (BEND) responding to interferon tau (IFNT).

Palma-Vera SE, Einspanier R - Reprod. Biol. Endocrinol. (2016)

Bottom Line: Bioinformatic analyses detected that PLA2G4A was associated to IRF-1 and IRF-6, while ESR1, PR and PTGS2 were associated to only IRF-6.All genes exhibit one motif per IRF, except miR-106a that had three binding sites for IRF-6.Future studies will use these data to detect new IFNT regulatory mechanisms in the endometrium.

View Article: PubMed Central - PubMed

Affiliation: Institute of Veterinary Biochemistry, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany.

ABSTRACT

Background: In ruminants, embryo implantation depends on progesterone (P4) and interferon tau (IFNT) controlling endometrial function. IFNT antagonizes bovine endometrial cells (BEND) response to phorbol 12,13-dibutyrate (PDBU) through posttranscriptional regulation of gene expression. We have previously described microRNAs (miRNAs) profiles in bovine endometrium, detecting miR-106a, relevant for embryo maternal communication. In this study, we investigated the expression miR-106a and genes for prostaglandin-endoperoxide synthase 2 (PTGS2), phospholipase A2, group IVA (PLA2G4A), estrogen receptor 1 (ESR1) and progesterone receptor (PR) in response to IFNT in BEND cells and searched for interferon responsive factors (IRFs) binding sites in their promoter genomic regions. The aim of this study was to unravel the molecular mechanisms involved in IFNT signalling and its regulation of miR-106a.

Findings: PTGS2 showed increased expression under PDBU, which was antagonized by IFNT. IFNT induced expression of PR and miR-106a and downregulation of ESR1 and PR. Bioinformatic analyses detected that PLA2G4A was associated to IRF-1 and IRF-6, while ESR1, PR and PTGS2 were associated to only IRF-6. All genes exhibit one motif per IRF, except miR-106a that had three binding sites for IRF-6.

Conclusions: We report the IFNT regulatory effect on miR-106a expression through IRF-6 in bovine endometrial cells. We identified a set of potential binding sites for IRF-1 and IRF-6 within the bovine genome. A set of candidate gene regions could be characterized where IFNT can act via IRFs to regulate the expression of proteins and miRNAs. Future studies will use these data to detect new IFNT regulatory mechanisms in the endometrium.

No MeSH data available.


Related in: MedlinePlus

Regulation of PTGS2, PLA2G4A, ESR1 and PR expression in BEND cells. a Normalized log 2 fold change mRNA expression of characteristic genes for BEND cells in response to treatment with PDBU, IFNT, P4 and their combinations. Transcript expression was normalized to a combination of two housekeeping genes (SUZ12 and SDHA). b Matrix of significances (white: p < 0.05; black: p > 0.05). For each experiment, six biological replicates were used. Outliers are indicated as single dots above or below the whiskers
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Fig1: Regulation of PTGS2, PLA2G4A, ESR1 and PR expression in BEND cells. a Normalized log 2 fold change mRNA expression of characteristic genes for BEND cells in response to treatment with PDBU, IFNT, P4 and their combinations. Transcript expression was normalized to a combination of two housekeeping genes (SUZ12 and SDHA). b Matrix of significances (white: p < 0.05; black: p > 0.05). For each experiment, six biological replicates were used. Outliers are indicated as single dots above or below the whiskers

Mentions: Previous studies have described the antagonizing effect of IFNT when PDBU was added to BEND cells. The result was a reduction of the mRNA of PTGS2 and PLA2G4A [8, 9, 11]. In our study, PTGS2 and PLA2G4A were upregulated by PDBU. For PTGS2, the PDBU effect was antagonized by IFNT, but this was not observed for PLA2G4A (Fig. 1). The lack of PLA2G4A regulation implies a stronger effect of IFNT on the expression of PTGS2 and a reduced effect on the expression of PLA2G4A. It has been shown that IFNT antagonizes the effect PDBU on the protein levels of PLA2G4A [9]. Thus, it could be possible that at the mRNA level, this effect remains inconspicuous. Nevertheless, the downregulation of PTGS2 corroborates the validity of our assays.Fig. 1


Experimental and bioinformatic analysis of cultured Bovine Endometrial Cells (BEND) responding to interferon tau (IFNT).

Palma-Vera SE, Einspanier R - Reprod. Biol. Endocrinol. (2016)

Regulation of PTGS2, PLA2G4A, ESR1 and PR expression in BEND cells. a Normalized log 2 fold change mRNA expression of characteristic genes for BEND cells in response to treatment with PDBU, IFNT, P4 and their combinations. Transcript expression was normalized to a combination of two housekeeping genes (SUZ12 and SDHA). b Matrix of significances (white: p < 0.05; black: p > 0.05). For each experiment, six biological replicates were used. Outliers are indicated as single dots above or below the whiskers
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4835850&req=5

Fig1: Regulation of PTGS2, PLA2G4A, ESR1 and PR expression in BEND cells. a Normalized log 2 fold change mRNA expression of characteristic genes for BEND cells in response to treatment with PDBU, IFNT, P4 and their combinations. Transcript expression was normalized to a combination of two housekeeping genes (SUZ12 and SDHA). b Matrix of significances (white: p < 0.05; black: p > 0.05). For each experiment, six biological replicates were used. Outliers are indicated as single dots above or below the whiskers
Mentions: Previous studies have described the antagonizing effect of IFNT when PDBU was added to BEND cells. The result was a reduction of the mRNA of PTGS2 and PLA2G4A [8, 9, 11]. In our study, PTGS2 and PLA2G4A were upregulated by PDBU. For PTGS2, the PDBU effect was antagonized by IFNT, but this was not observed for PLA2G4A (Fig. 1). The lack of PLA2G4A regulation implies a stronger effect of IFNT on the expression of PTGS2 and a reduced effect on the expression of PLA2G4A. It has been shown that IFNT antagonizes the effect PDBU on the protein levels of PLA2G4A [9]. Thus, it could be possible that at the mRNA level, this effect remains inconspicuous. Nevertheless, the downregulation of PTGS2 corroborates the validity of our assays.Fig. 1

Bottom Line: Bioinformatic analyses detected that PLA2G4A was associated to IRF-1 and IRF-6, while ESR1, PR and PTGS2 were associated to only IRF-6.All genes exhibit one motif per IRF, except miR-106a that had three binding sites for IRF-6.Future studies will use these data to detect new IFNT regulatory mechanisms in the endometrium.

View Article: PubMed Central - PubMed

Affiliation: Institute of Veterinary Biochemistry, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany.

ABSTRACT

Background: In ruminants, embryo implantation depends on progesterone (P4) and interferon tau (IFNT) controlling endometrial function. IFNT antagonizes bovine endometrial cells (BEND) response to phorbol 12,13-dibutyrate (PDBU) through posttranscriptional regulation of gene expression. We have previously described microRNAs (miRNAs) profiles in bovine endometrium, detecting miR-106a, relevant for embryo maternal communication. In this study, we investigated the expression miR-106a and genes for prostaglandin-endoperoxide synthase 2 (PTGS2), phospholipase A2, group IVA (PLA2G4A), estrogen receptor 1 (ESR1) and progesterone receptor (PR) in response to IFNT in BEND cells and searched for interferon responsive factors (IRFs) binding sites in their promoter genomic regions. The aim of this study was to unravel the molecular mechanisms involved in IFNT signalling and its regulation of miR-106a.

Findings: PTGS2 showed increased expression under PDBU, which was antagonized by IFNT. IFNT induced expression of PR and miR-106a and downregulation of ESR1 and PR. Bioinformatic analyses detected that PLA2G4A was associated to IRF-1 and IRF-6, while ESR1, PR and PTGS2 were associated to only IRF-6. All genes exhibit one motif per IRF, except miR-106a that had three binding sites for IRF-6.

Conclusions: We report the IFNT regulatory effect on miR-106a expression through IRF-6 in bovine endometrial cells. We identified a set of potential binding sites for IRF-1 and IRF-6 within the bovine genome. A set of candidate gene regions could be characterized where IFNT can act via IRFs to regulate the expression of proteins and miRNAs. Future studies will use these data to detect new IFNT regulatory mechanisms in the endometrium.

No MeSH data available.


Related in: MedlinePlus