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REV-ERBα inhibits the PTGS2 expression in bovine uterus endometrium stromal and epithelial cells exposed to ovarian steroids.

Isayama K, Chen H, Yamauchi N, Hattori MA - J. Reprod. Dev. (2014)

Bottom Line: BMAL1 knockdown with specific siRNA caused a significant decrease in the transcript levels of NR1D1 and PTGS2 in USCs, but not in UECs.The production of PGF2α also decreased in USCs after BMAL1 knockdown, while its level did not significantly change in UECs.Collectively, these results indicate that REV-ERBα plays an inhibitory role in the expression of PTGS2 in both bovine USCs and UECs treated with ovarian steroids.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal and Marine Bioresource Sciences, Graduate School of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.

ABSTRACT
The nuclear receptor REV-ERBα (encoded by NR1D1) has a critical role in metabolism and physiology as well as circadian rhythm. Here, we investigated the possible contribution of clock genes including NR1D1 to the secretion of prostaglandin F2α (PGF2α) from bovine uterine stromal (USCs) and epithelial cells (UECs) by modulating the expression of PTGS2. The circadian oscillation of clock genes in the cells was weak compared with that reported in rodents, but the expression of BMAL1, PER1, and NR1D1 was changed temporally by treatment with ovarian steroids. Significant expression of clock genes including NR1D1 was detected in USCs exposed to progesterone. NR1D1 was also significantly expressed in UECs exposed to estradiol. The expression of PTGS2 was suppressed in USCs exposed to progesterone, while the expression was initially suppressed in UECs exposed to estradiol and then increased after long-term exposure to estradiol. BMAL1 knockdown with specific siRNA caused a significant decrease in the transcript levels of NR1D1 and PTGS2 in USCs, but not in UECs. The production of PGF2α also decreased in USCs after BMAL1 knockdown, while its level did not significantly change in UECs. The transcript level of PTGS2 was increased by treatment with the antagonist of REV-ERBα in both cell types, but the agonist was ineffective. In these two cell types treated with the agonist or antagonist, the PGF2α production coincided well with the PTGS2 expression. Collectively, these results indicate that REV-ERBα plays an inhibitory role in the expression of PTGS2 in both bovine USCs and UECs treated with ovarian steroids.

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Generation of bioluminescence oscillations by rat and bovine USCs transfected with pGL3 vector containing the mousePer1 promoter region after synchronization with forskolin. (A) Schematic of the pGL3 vectorcontaining the mouse Per1 promoter region (upper). Black bars indicate the location of E-box motifs(–146 to –151, –509 to –514, and –1255 to –1260) and a cyclic-AMP response element (CRE, –1725 to –1732). (B)Bioluminescence activity was induced in bovine and rat USCs transfected with 1 μg of the constructed vector bysynchronization with forskolin. Bioluminescence was monitored in real time in serum-free medium DMEM/F12supplemented with 0.1 mM luciferin, 0.1% BSA, 1% ITS, 1×AA and 100 nM P4. Each value represents the meansof three independent determinations.
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fig_001: Generation of bioluminescence oscillations by rat and bovine USCs transfected with pGL3 vector containing the mousePer1 promoter region after synchronization with forskolin. (A) Schematic of the pGL3 vectorcontaining the mouse Per1 promoter region (upper). Black bars indicate the location of E-box motifs(–146 to –151, –509 to –514, and –1255 to –1260) and a cyclic-AMP response element (CRE, –1725 to –1732). (B)Bioluminescence activity was induced in bovine and rat USCs transfected with 1 μg of the constructed vector bysynchronization with forskolin. Bioluminescence was monitored in real time in serum-free medium DMEM/F12supplemented with 0.1 mM luciferin, 0.1% BSA, 1% ITS, 1×AA and 100 nM P4. Each value represents the meansof three independent determinations.

Mentions: To investigate whether the cellular clockwork functions in bovine uterus cells, we first analyzed mouse Per1promoter activity as an indicator of the clockwork. There are three E-box sites and one CRE in the mousePer1 promoter region (Fig. 1AFig. 1.


REV-ERBα inhibits the PTGS2 expression in bovine uterus endometrium stromal and epithelial cells exposed to ovarian steroids.

Isayama K, Chen H, Yamauchi N, Hattori MA - J. Reprod. Dev. (2014)

Generation of bioluminescence oscillations by rat and bovine USCs transfected with pGL3 vector containing the mousePer1 promoter region after synchronization with forskolin. (A) Schematic of the pGL3 vectorcontaining the mouse Per1 promoter region (upper). Black bars indicate the location of E-box motifs(–146 to –151, –509 to –514, and –1255 to –1260) and a cyclic-AMP response element (CRE, –1725 to –1732). (B)Bioluminescence activity was induced in bovine and rat USCs transfected with 1 μg of the constructed vector bysynchronization with forskolin. Bioluminescence was monitored in real time in serum-free medium DMEM/F12supplemented with 0.1 mM luciferin, 0.1% BSA, 1% ITS, 1×AA and 100 nM P4. Each value represents the meansof three independent determinations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219993&req=5

fig_001: Generation of bioluminescence oscillations by rat and bovine USCs transfected with pGL3 vector containing the mousePer1 promoter region after synchronization with forskolin. (A) Schematic of the pGL3 vectorcontaining the mouse Per1 promoter region (upper). Black bars indicate the location of E-box motifs(–146 to –151, –509 to –514, and –1255 to –1260) and a cyclic-AMP response element (CRE, –1725 to –1732). (B)Bioluminescence activity was induced in bovine and rat USCs transfected with 1 μg of the constructed vector bysynchronization with forskolin. Bioluminescence was monitored in real time in serum-free medium DMEM/F12supplemented with 0.1 mM luciferin, 0.1% BSA, 1% ITS, 1×AA and 100 nM P4. Each value represents the meansof three independent determinations.
Mentions: To investigate whether the cellular clockwork functions in bovine uterus cells, we first analyzed mouse Per1promoter activity as an indicator of the clockwork. There are three E-box sites and one CRE in the mousePer1 promoter region (Fig. 1AFig. 1.

Bottom Line: BMAL1 knockdown with specific siRNA caused a significant decrease in the transcript levels of NR1D1 and PTGS2 in USCs, but not in UECs.The production of PGF2α also decreased in USCs after BMAL1 knockdown, while its level did not significantly change in UECs.Collectively, these results indicate that REV-ERBα plays an inhibitory role in the expression of PTGS2 in both bovine USCs and UECs treated with ovarian steroids.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal and Marine Bioresource Sciences, Graduate School of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.

ABSTRACT
The nuclear receptor REV-ERBα (encoded by NR1D1) has a critical role in metabolism and physiology as well as circadian rhythm. Here, we investigated the possible contribution of clock genes including NR1D1 to the secretion of prostaglandin F2α (PGF2α) from bovine uterine stromal (USCs) and epithelial cells (UECs) by modulating the expression of PTGS2. The circadian oscillation of clock genes in the cells was weak compared with that reported in rodents, but the expression of BMAL1, PER1, and NR1D1 was changed temporally by treatment with ovarian steroids. Significant expression of clock genes including NR1D1 was detected in USCs exposed to progesterone. NR1D1 was also significantly expressed in UECs exposed to estradiol. The expression of PTGS2 was suppressed in USCs exposed to progesterone, while the expression was initially suppressed in UECs exposed to estradiol and then increased after long-term exposure to estradiol. BMAL1 knockdown with specific siRNA caused a significant decrease in the transcript levels of NR1D1 and PTGS2 in USCs, but not in UECs. The production of PGF2α also decreased in USCs after BMAL1 knockdown, while its level did not significantly change in UECs. The transcript level of PTGS2 was increased by treatment with the antagonist of REV-ERBα in both cell types, but the agonist was ineffective. In these two cell types treated with the agonist or antagonist, the PGF2α production coincided well with the PTGS2 expression. Collectively, these results indicate that REV-ERBα plays an inhibitory role in the expression of PTGS2 in both bovine USCs and UECs treated with ovarian steroids.

Show MeSH
Related in: MedlinePlus