A microarray analysis for genes regulated by interferon- in ovine luminal epithelial cells (original) (raw)
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Biology of Reproduction, 1999
Oxytocin (OT) is responsible for the episodic release of luteolytic prostaglandin (PG) F 2␣ from the uterus in ruminants. The attenuation of OT-stimulated uterine PGF 2␣ secretion by interferon-(IFN-) is essential for prevention of luteolysis during pregnancy in cows. To better understand the mechanisms involved, the effect of recombinant bovine IFN-(rbIFN-) on OTinduced PG production and cyclooxygenase-2 (COX-2) and PGF synthase (PGFS) expression in cultured endometrial epithelial cells was investigated. Cells were obtained from cows at Days 1-3 of the estrous cycle and cultured to confluence in RPMI medium supplemented with 5% steroid-free fetal calf serum. The cells were then incubated in the presence or absence of either 100 ng/ml OT or OT؉100 ng/ml rbIFN-for 3, 6, 12, and 24 h. OT significantly increased PGF 2␣ and PGE 2 secretion at all time points (p Ͻ 0.01), while rbIFN-inhibited the OTinduced PG production and reduced OT receptor binding in a time-dependent manner. OT increased the steady-state level of COX-2 mRNA, measured by Northern blot, which was maximal at 3 h (9-fold increase) and then decreased with time (p Ͻ 0.01). OT also caused an increase in COX-2 protein, which peaked at 12 h (11-fold increase), as measured by Western blot. Addition of rbIFN-suppressed the induction of COX-2 mRNA (89%, p Ͻ 0.01) and COX-2 protein (50%, p Ͻ 0.01) by OT. OT also increased PGFS mRNA, and this stimulation was attenuated by rbIFN-(p Ͻ 0.01). To ensure that the decrease in COX-2 was not solely due to down-regulation of the OT receptor, cells were stimulated with a phorbol ester (phorbol 12-myristate 13-acetate; PMA) in the presence and absence of rbIFN-. The results showed that rbIFN-also decreased PMA-stimulated PG production and COX-2 protein. It can be concluded that rbIFNinhibition of OT-stimulated PG production is due to down-regulation of OT receptor, COX-2, and PGFS.
Reproductive Biology and Endocrinology, 2003
In sheep, the uterus produces luteolytic pulses of prostaglandin F 2α (PGF) on Days 15 to 16 of estrous cycle to regress the corpus luteum (CL). These PGF pulses are produced by the endometrial lumenal epithelium (LE) and superficial ductal glandular epithelium (sGE) in response to binding of pituitary and/or luteal oxytocin to oxytocin receptors (OTR) and liberation of arachidonic acid, the precursor of PGF. Cyclooxygenase-one (COX-1) and COX-2 are rate-limiting enzymes in PGF synthesis, and COX-2 is the major form expressed in ovine endometrium. During pregnancy recognition, interferon tau (IFNτ), produced by the conceptus trophectoderm, acts in a paracrine manner to suppress development of the endometrial epithelial luteolytic mechanism by inhibiting transcription of estrogen receptor α (ERα) (directly) and OTR (indirectly) genes.
Biology of Reproduction, 2001
Interferon tau (IFNtau) is the signal for maternal recognition of pregnancy in ruminants. The positive effects of IFNtau on IFN-stimulated gene (ISG) expression are mediated by ISG factor 3 (ISGF3), which is composed of signal transducer and activator of transcription (Stat) 1, Stat 2, and IFN regulatory factor-9 (IRF-9), and by gamma-activated factor (GAF), which is a Stat 1 homodimer. Induction of ISGs, such as ISG17 and 2',5'-oligoadenylate synthetase, by IFNtau during pregnancy is limited to the endometrial stroma (S) and glandular epithelium (GE) of the ovine uterus. The IRF-2, a potent transcriptional repressor of ISG expression, is expressed in the luminal epithelium (LE). This study determined effects of the estrous cycle, pregnancy, and IFNtau on expression of Stat 1, Stat 2, IRF-9, IRF-1, and IRF-2 genes in the ovine endometrium. In cyclic ewes, Stat 1, Stat 2, IRF-1, and IRF-9 mRNA and protein were detected at low levels in the S and GE. During pregnancy, expression of these genes increased only in the S and GE. Expression of IRF-2 was detected only in the LE and superficial GE (sGE) of both cyclic and pregnant ewes. In cyclic ewes, intrauterine administration of IFNtau stimulated Stat 1, Stat 2, IRF-9, and IRF-1 expression in the endometrium. Ovine IRF-2 repressed transcriptional activity driven by IFN-stimulated response elements that bind ISGF3, but not by gamma-activation sequences that bind GAF. These results suggest that IRF-2 in the LE and sGE restricts IFNtau induction of ISGs to the S and GE. In the S and GE, IFNtau hyperactivation of ISG expression likely involves formation and actions of the transcription factors ISGF3 and, perhaps, IRF-1.
Domestic Animal Endocrinology, 2002
Luteal regression is caused by a pulsatile release of prostaglandin (PG) F 2␣ from the uterus in the late luteal phase in most mammals including cattle. Although it has been proposed in ruminants that pulsatile PGF 2␣ secretion is generated by a positive feedback loop between luteal and/or hypophyseal oxytocin and uterine PGF 2␣ , the bovine endometrium may possess other mechanisms for initiation of luteolytic PGF 2␣ secretion. It has been recently demonstrated that tumor necrosis factor-␣ (TNF-␣) stimulates PGF 2␣ output from bovine endometrial tissue not only during the follicular phase but also during the late luteal phase, suggesting that TNF-␣ is a factor in the initiation of luteolysis in cattle. Furthermore, our recent study has shown that IFN-suppresses the action of TNF-␣ on PGF 2␣ synthesis by the bovine endometrium in vitro, suggesting that IFN-plays a luteoprotective role by inhibiting TNF-␣-induced PGF 2␣ production in early pregnancy. On the other hand, factors other than oxytocin or TNF-␣ have also been suggested to be involved in the regulation of PGF 2␣ synthesis by bovine endometrium. The purpose of this review is to summarize our current understanding of the endocrine mechanisms that regulate the timing and pattern of uterine PGF 2␣ secretion during the estrous cycle and early pregnancy.
Biology of Reproduction, 2003
Tumor necrosis factor-␣ (TNF␣) has been shown to be a potent stimulator of prostaglandin (PG) F 2␣ synthesis in bovine endometrial stromal cells. The aims of the present study were to determine the effect of interferon-(IFN) on TNF␣-stimulated PGF 2␣ synthesis and the intracellular mechanisms of TNF␣ and IFN action in the stromal cells. When cultured bovine stromal cells were exposed to TNF␣ (0.006-0.6 nM) for 24 h, the production of PGF 2␣ and cyclooxygenase (COX)-2 gene expression were stimulated by TNF␣ (0.06-0.6 nM, P Ͻ 0.05). Moreover, a specific COX-2 inhibitor (NS-398; 5 nM) blocked the stimulatory effect of TNF␣ on PGF 2␣ production (P Ͻ 0.05). Although IFN (0.03-30 ng/ml) did not stimulate basal PGF 2␣ production in the stromal cells, it suppressed TNF␣ action in PGF 2␣ production dose dependently (P Ͻ 0.05). Moreover, the stimulatory effect of TNF␣ (0.6 nM) on COX-2 gene expression was completely blocked by IFN (30 ng/ml; P Ͻ 0.05), although the gene expression of COX-2 was not influenced by IFN. The overall results indicate that the stimulatory effect of TNF␣ on PGF 2␣ production is mediated by the up-regulation of COX-2 gene expression and suggest that one of the mechanisms of the inhibitory effect of IFN on luteolysis is the inhibition of TNF␣ action in PGF 2␣ production in the stromal cells by the down-regulation of COX-2 gene expression stimulated by TNF␣.
Endocrinology, 1998
Estradiol (E2) and progesterone are responsible for regulating PG synthesis in the endometrium during the estrous cycle and interferon-(IFN-) alters PG synthesis during early pregnancy in ruminants. In this study, we examined the effects of these steroid hormones and recombinant bovine IFN-(rbIFN-) on PG production and on cyclooxygenase-2 (COX-2) and PG F (PGF) synthase (PGFS) gene expression in isolated endometrial cells. E2 decreased both PGF 2␣ and PG E 2 (PGE 2) whereas progesterone increased PGF 2␣ secretion in epithelial cells. Steroid hormones had no effect on PG production in stromal cells. rbIFN-attenuated both PGF 2␣ and PGE 2 production in epithelial cells and enhanced their production, and the ratio of PGE 2 to PGF 2␣ , in stromal cells. Northern blot analysis showed that E2 and rbIFN-decreased COX-2 messenger RNA (mRNA) levels in epithelial cells. Conversely, rbIFN-increased COX-2 mRNA in stromal cells. Furthermore, rbIFN-decreased PGFS mRNA in both cell types and this was associated with the increase in PGE 2 /PGF 2␣ ratio. These results show that the regulation of PG synthesis by steroid hormones is different in endometrial epithelial and stromal cells in vitro. The attenuation of PGF 2␣ secretion from epithelial cells and increased PGE 2 production in stromal cells by rbIFN-are modulated by steroid hormones.
Possible role of IFNT on the bovine corpus luteum and neutrophils during the early pregnancy
Reproduction (Cambridge, England), 2015
When pregnancy is established, interferon tau (IFNT), a well-known pregnancy recognition signal in ruminants, is secreted by embryonic trophoblast cells and acts within the uterus to prepare for pregnancy. IFNT acts as an endocrine factor on the corpus luteum (CL) to induce refractory ability against the luteolytic action of PGF2a. Hypothesising that IFNT may influence not only the uterine environment but also the CL in cows via local or peripheral circulation, we investigated qualitative changes in the CL of pregnant cows during the maternal recognition period (day 16) and the CL of non-pregnant cows. The CL of pregnant animals had higher number of neutrophils, and the expression of interleukin (IL)-8 mRNA and its protein was higher as well, as compared with the CL of non-pregnant animals. Although IFNT did not affect progesterone secretion and neutrophil migration directly, it stimulated IL-8 mRNA expression on luteal cells influenced the neutrophils, resulting in the increased mi...
Endocrinology, 2008
The ruminant conceptus synthesizes and secretes interferon (IFN)-, which presumably acts via an intrauterine paracrine mechanism to signal maternal recognition of pregnancy. The aims of this study were to determine whether IFN-stimulated genes (ISG) such as ISG15 and OAS-1 are differentially expressed in blood cells circulating in the uterus of ewes; whether extrauterine components of the reproductive tract such as the corpus luteum (CL) also express mRNA for these ISG, and whether antiviral activity is greater in uterine vein than in uterine artery during early pregnancy. The concentrations of mRNA for both ISG were significantly greater (P < 0.0001) in endometrium and jugular blood of 15-d pregnant ewes than in nonpregnant ewes. ISG15 and OAS-1 mRNA con-centrations were also greater (P < 0.05) in CL from 15-d pregnant ewes than in nonpregnant ewes. Immunohistochemistry revealed intense staining for ISG15 in large luteal cells on d 15 of pregnancy. Blood cells from uterine artery and vein of 15-d pregnant ewes had similar ISG15 and OAS-1 mRNA concentrations, suggesting that these cells were not conditioned by IFN-within the uterus. By using an antiviral assay, uterine venous blood was found to contain 500-to 1000-fold higher concentrations of bioactive IFN-than in uterine arterial blood on d 15 of pregnancy. It is concluded that uterine vein releases IFN-, which induces ISG in extrauterine tissues such as the CL during the time of maternal recognition of pregnancy.
Journal of Dairy Science, 2010
Interferon-τ (IFNT), the pregnancy recognition signal in ruminant species, is secreted by conceptus trophectoderm cells and induces expression of IFN-stimulated gene 15 (ISG15) in the uterus and corpus luteum (CL) in ewes. Expression of ISG15 in ovine CL is speculated to be through an endocrine pathway, but it is unclear whether expression of ISG15 in bovine CL is via such a pathway. In this study, CL were obtained from cows on d 16, 25, 60, 120, 180, and 270 of pregnancy, and endometrium, mammary gland, ovarian stroma, and CL were also collected from cows on d 18 of pregnancy and on d 15 and 18 of the estrous cycle. All tissue explants from d 15 of the estrous cycle were cultured in the absence or presence of 100 ng/mL of recombinant bovine IFNT for 24 h. The results indicated that ISG15 and conjugated proteins were expressed in CL of both cyclic and pregnant cows regardless of pregnancy status and were up-regulated during early pregnancy. The mammary gland from d 18 of pregnancy did not express ISG15, but explants of the mammary gland from d 15 of the estrous cycle did express ISG15 after being treated with IFNT. However, luteal explants from d 15 of the estrous cycle did not express ISG15 after being cultured for 24 h. In conclusion, ISG15 expression is up-regulated in the bovine CL during early pregnancy. Interestingly, cultured CL cells do not respond to IFNT, suggesting that the pregnancy-dependent stimulation of ISG15 expression is controlled by something other than IFNT in the bloodstream.