Expression and regulation of Foxa2 in the rat uterus during early pregnancy

The forkhead box a (Foxa) protein family has been found to play important roles in mammals. Recently, the expression of Foxa2 was reported in the mouse uterus, and it was reported to be involved in regulation of implantation. However, the regulation of Foxa2 expression in the uterus is still poorly understood. Therefore, the present study was conducted to investigate the expressional profiles of Foxa2 in the rat uterus during the estrus cycle and pregnancy. Furthermore, the effect of steroid hormones and Hedgehog protein on the expression of Foxa2 was analyzed in vivo and in vitro. In this study, the level of expression of Foxa2 was low in the rat uterus during the different stages of the estrus cycle. However, the expression increased transiently during early pregnancy at 3.5 days post coitus (dpc) and decreased at 5.5 dpc. In ovariectomized rats, P4 treatment had no effect on the expression of Foxa2 compared with the expression in control animals. Moreover, the expression of Foxa2 in cultured epithelial cells was not increased by P4 treatment in vitro. However, Foxa2 expression was significantly decreased in the rat uterus after 24 h of E2 treatment. Treatment of cells with a recombinant Hedgehog protein significantly increased the expression of Foxa2. These results suggest that the expression of Foxa2 may transiently increase just before the implantation and it may be regulated by E2 and Hedgehog protein.     

交感神经对妊娠早期大鼠子宫内DKK-1和Wnt-1表达的影响

为研究交感神经对妊娠早期子宫内DKK-1和Wnt-1的调控作用,选取180~200日龄健康的性成熟Wistar雌性大鼠,随机分为对照组和处理组,处理组采用静脉注射6-OHDA损毁交感神经,用免疫组化法和qRT-PCR法,观察和分析交感神经对妊娠早期大鼠子宫内膜的DKK-1和Wnt-1蛋白及mRNA相对表达量的影响。结果显示:6-OHDA损毁交感神经后,对卵巢排卵数无显著性影响,但子宫囊胚数量显著下降、囊胚发育延迟,导致子宫腔上皮和腺上皮细胞自分泌DKK-1和Wnt-1紊乱,呈现与正常状态相反的变化趋势。结果表明:在着床窗口期,交感神经通过影响DKK-1对Wnt-1信号分子的调控作用而影响囊胚着床、发育。     

Expression patterns of the implantation-associated genes in the uterus during the estrous cycle in mice

The mRNA expression patterns of EGF, HB-EGF, Amphiregulin, EGF receptor, IGF-1, CSF-1, IL-1 alpha, IL-1 beta, IL-1 receptor type 1, IL-1 receptor antagonist, LIF, COX-1, COX-2, Mucin-1, calcitonin, and rat USAG-1 mouse homologue, all of which are involved in the process of conceptus implantation to the endometrium, were examined during the estrous cycle by means of real-time quantitative PCR. COX-2, HB-EGF, LIF, Mucin-1, CSF-1, IL-1 alpha, IL-1 beta, and IL-1 receptor antagonist were temporally regulated during the estrous cycle and highly expressed during the estrous stage. In the case of COX-1, EGF, IGF-1, and EGF receptor, the highest mRNA expression was during the diestrous stage. In contrast, the rat USAG-1 mouse homologue mRNA expression did not change during the estrous cycle. These results indicate that rat USAG-1 mouse homologue expression at implantation might be specifically regulated by embryonic factors rather than the maternal environment.     

The uterus and early pregnancy failure in the mare

The environment the equine conceptus finds itself in when it arrives in the uterus some 6 days after ovulation will determine if it will thrive or die. The uterus will be its home for approximately the next 11 months and as such it needs to both coordinate the growth of the placenta and ensure there is enough nourishment passing to this organ for the fetus to develop normally. The trophoblast, endometrium, maternal ovaries and, later in pregnancy, the fetal gonads, all play roles in the hormonal changes that orchestrate these events. Although failures of these processes later in pregnancy can have catastrophic effects for the fetus, it is in early gestation that the foundations for a successful pregnancy are laid. This paper therefore concentrates on some of the noninfectious influences the uterus may have on survival of the young conceptus.     

Expression and regulation of Enpp2 in rat uterus during the estrous cycle

Ectonucleotide pyrophosphatase/phosphodiestrase 2 (Enpp2) isolated from the supernatant of human melanoma cells is a lysophospholipase D that transforms lysophosphatidylcholine into lysophospatidic acid. Although multiple analyses have investigated the function of Enpp2 in the hypothalamus, its role in the uterus during the estrous cycle is not well understood. In the present study, rat uterine Enpp2 was analyzed by RT-PCR, Western blotting, and immunohistochemistry. Quantitative PCR analysis demonstrated that uterine Enpp2 mRNA was decreased during estrus compared to proestrus and diestrus. To determine whether uterine Enpp2 expression is affected by sex steroid hormones, immature rats were treated with 17β-estradiol (E2), progesterone, or both on postnatal days 14 to 16. Interestingly, the expression of Enpp2 mRNA and protein were down-regulated by E2 in the uterus during estrus but not during proestrus or diestrus, suggesting that Enpp2 may play a role in uterine function during estrus. Enpp2 is primarily localized in the stromal cells of the endometrium during proestrus and estrus. During diestrus, Enpp2 was highly expressed in the epithelial cells of the endometrium. Taken together, these results suggest that uterine Enpp2 may be regulated by E2 and plays a role in reproductive functions during female rat development.     

Progesterone receptor mRNA levels during pregnancy, labor, lactation and the estrous cycle in rat uterus

Progesterone plays important roles in the regulation of female reproduction. In this study, progesterone receptor (PR) mRNA levels in rat uterus during pregnancy, labor, lactation and the estrous cycle were examined by competitive RT-PCR. During pregnancy and lactation, PR mRNA levels had decreased on day 20 of pregnancy (P20) and P21 compared with P15 but increased during labor. After a decline on day 1 of lactation (L1), PR mRNA levels had increased again on L3 and L14 compared with P15, P18, P20, P21 and P21pm (at 2200-2300 h on P21). There was no significant change in the PR mRNA level during the estrous cycle. The PR mRNA level did not change during 1 week of progesterone treatment or afterwards. Injection of 17beta-estradiol did not affect PR mRNA levels in rats treated with progesterone or those without any injections. In rats on P18, 17beta-estradiol injection did not change PR mRNA levels after sham-operation but induced an increase in PR mRNA levels of rats ovariectomized 6 h before the treatment. These results suggest that uterine PR mRNA levels are differently regulated during late pregnancy, labor and lactation, and during labor estrogen is one of the essential factors for the increase in PR mRNA levels.     

Expression of IFNAR1 and IFNAR2 in cattle placenta during early pregnancy

Interferon‐tau (IFNT), a type I interferon, is an antiluteolytic factor secreted by trophoderm during pregnancy. IFNT transmitted signals or stimulated the expression of some factors to build maternal recognition and keep pregnancy by binding its receptors, IFNT receptor 1(IFNAR1) and IFNT receptor 2 (IFNAR2). Up to now, the expression model and roles of IFNAR1 and IFNAR2 in placenta have not been investigated in cattle. In this study, the localization and expression of IFNAR1 and IFNAR2 in the cattle placenta at days 18–50 of pregnancy were detected by histological examination, immunofluorescence staining and real‐time qPCR. The results showed that IFNAR1 mainly distributed in chorioallantoic membrane, endometrial epithelium, cotyledon and caruncle during the early pregnancy of cattle with change in time‐ and position‐dependent. IFNAR1 and IFNAR2 mRNA expression were mainly detected in chorioallantoic membrane and cotyledon, and markedly increased along with pregnancy process. Moreover, the mRNA expression level of IFNAR1 in chorioallantoic membrane and cotyledon was higher than that of IFNAR2. IFNAR mRNA was also expressed in caruncle tissues, which experienced a tendency of decrease from days 21 to 36, followed by increase after days 36. These results provide morphological basis and quantitative data for investigating the roles of IFNAR1 and IFNAR2 on development of cattle placenta and pregnancy maintenance.     

Expression dynamics of bovine MX genes in the endometrium and placenta during early to mid pregnancy

MX belongs to a family of type I interferon (IFN)-stimulated genes, and the MX protein has antiviral activity. MX has at least two isoforms, known as MX1 and MX2, in mammals. Moreover, bovine MX1 has been found to have alternative splice variants—namely, MX1-a and MX1B. In ruminants, IFN-τ—a type I IFN—is temporarily produced from the conceptus before implantation and induces MX expression in the endometrium. However, the expression dynamics of MX after implantation are not clear. In the present study, we investigated the expression of MX1-a, MX1B and MX2 in the endometrium and placenta before and after implantation along with the expression of IFN-α, type I receptors (IFNAR1 and IFNAR2) and interferon regulatory factors (IRF3 and IRF9). Pregnant uterine samples were divided into five groups according to pregnancy days 14–18, 25–40, 50–70, 80–100, and 130–150. Tissue samples were collected from the intercaruncular endometrium (IC), caruncular endometrium (C) and fetal placenta (P). Although all the MX expressions were significantly higher in the IC and C at days 14–18, presumably caused by embryo-secreted IFN-τ stimulation, their expressions were also detectable in the IC, C and P after implantation. Furthermore, IFN-α expression was significantly higher in the IC. RT-PCR indicated IFNAR1, IFNAR2, IRF3 and IRF9 mRNA in all the tissues during pregnancy. These results suggest that all the MX genes are affected by the type I IFN pathway during pregnancy and are involved in an immune response to protect the mother and fetus.     

Strangulation of the duodenum by the uterus during late pregnancy in two cows.

Two Swiss Braunvieh cows in late pregnancy underwent surgery because of a rare form of ileus due to strangulation of the duodenum at its caudal flexure by the gravid uterus. The whole uterus had passed through a gap between the mesoduodenum and duodenum and with increasing weight had led to strangulation of the duodenum. This was possible since the mesoduodenum and both walls of the greater omentum adjacent to its caudal edge were not connected with the duodenum, probably due to a congenital inhibitory malformation. A transsection and an end-to-end anastomosis of the duodenum were necessary in both cases since it was impossible to retract the gravid uterus through the defect. Postoperative recovering was uneventful in both cows, which were discharged after seven and five days respectively and calved normally about two months later. Copyright Harcourt Publishers Ltd.     

Left abomasal displacement between the uterus and rumen during bovine twin pregnancy

Here, we describe two cases of left displaced abomasum (LDA) in Holstein cattle at 6 and 9 months of twin gestation. Clinical examination revealed signs of proximal ileus with marked abdominal distension, but no ping sounds. An unusually high position of the gravid uterine horn on the left side was observed with ultrasound. Left exploratory laparotomy confirmed that the abomasum was displaced to the left and entrapped between the rumen and twin gravid uterine horn. A left surgical approach was necessary to correct the condition. Both animals recovered and gave birth to healthy twins. The present cases indicate that the subomental position of a heavy twin gravid uterine horn is a possible mechanical cause of LDA.     

Upregulation of interferon-stimulated genes and interleukin-10 in peripheral blood immune cells during early pregnancy in dairy cows

In cows, interferon-tau (IFNT) regulates maternal recognition around days 15-19 after artificial insemination (AI). The present study hypothesized that if key target genes of IFNT are clearly upregulated in earlier stages of pregnancy, these genes could be use as indices of future pregnancy in cows. Therefore, we determined the expression of these genes in peripheral blood mononuclear leukocytes (PBMCs) and polymorphonuclear granulocytes (PMNs) during the maternal recognition period (MRP). Twenty multiparous Holstein cows were subjected to AI on day 0 and categorized into the following groups: pregnancy (Preg, n = 9), embryonic death (ED, n = 5) and non-pregnancy (NP, n = 6). Progesterone levels in the Preg group were higher than those in the NP group on days 12-21. ISG15 and OAS-1 (IFN-stimulated genes: ISGs) mRNA in PBMCs on day 8 was higher in the Preg group than in the NP group, and these mRNAs in PMNs was higher in the Preg group on day 5 than in the NP and ED groups. Interleukin-10 (IL-10, Th2 cytokine) mRNA expression increased on day 8 in the PBMCs of pregnant cows. Tumor necrosis factor α (TNFα, Th1 cytokine) mRNA expression was stable in all groups. In an in vitro cell culture experiment, IFNT stimulated mRNA expression of ISGs in both PBMCs and PMNs. IFNT stimulated IL-10 mRNA expression in PBMCs, whereas IFNT increased TNFα mRNA levels in PBMCs in vitro. The results suggest that ISGs and IL-10 could be responsive to IFNT before the MRP in peripheral blood immune cells and may be useful target genes for reliable indices of pregnancy before the MRP.     

山羊妊娠早期胎盘滋养层细胞TET1、MMPs与TIMPs的表达规律

TET1的去甲基化作用对胎盘功能具有重要调控作用,胎盘滋养层分泌的基质金属酶(matrix metalloproteinase,MMPs)使其具有迁移和侵袭功能,保证了滋养层细胞正常侵入子宫内膜建立妊娠,但滋养层细胞TET1、MMPs及TIMPs在山羊妊娠早期胎盘滋养层细胞中的表达规律未知。因此,本试验取妊娠20,25,30,45和60 d大足黑山羊胎盘滋养层,利用荧光定量PCR技术检测并分析了MMP2和MMP9、TIMP1和TIMP2基因相对表达规律,观察TET1和MMP9在各时期胎盘滋养层中的分布规律。结果显示,除20,25 d外,MMP9的表达量均高于MMP2,在30和60 d差异显著(P0.05),45 d时差异达到极显著(P0.01);除20 d外,TIMP1的表达量均高于TIMP2,且在30,45 d时具有显著性差异(P0.05)。免疫组化染色结果发现TET1在细胞核和细胞质中均有表达,MMP9则主要分布于细胞质和细胞膜,且两者表达变化存在一定相似性。结果表明,山羊妊娠25～60 d,可能由MMP9和TIMP1主要调控胎盘滋养层细胞迁移和侵袭,且在该过程中,TET1可能通过影响MMP9的表达,进而影响山羊胎盘滋养层细胞迁移和侵袭。     

Histomorphological changes in the oviduct epithelium of the rabbit during early pregnancy

In this study, we determined the histomorphological changes in different regions of the oviduct (fimbriae, infundibulum, ampullae and isthmus) during early pregnancy (days 1-4) of the rabbit. It was observed that the number of secretory cells (PAS-positive cells) diminished in the fimbriae during the first 4 days of pregnancy with the corresponding increase in the number of non-secretory cells (PAS-negative cells). In contrast, there was an increase in the number of PAS-positive cells in the isthmus during early pregnancy. In both the infundibulum and the ampullae the number of PAS-positive cells was increased on day 3. A significant decrease in the height of the epithelium was observed in the fimbriae during early pregnancy. On the contrary, a significant increase in this parameter was observed in the isthmus during the first 3 days of pregnancy. A slight decrease in the height of the epithelium was observed in the infundibulum on day 4 of pregnancy, while in the ampullae it was significantly increased on days 1 and 3. The overall results indicate that during early pregnancy there is a selective modification in the histomorphological characteristics of the different regions of the rabbit oviduct.     

Characterization of effects of zearalenone in swine during early pregnancy

Mature gilts (n = 16) were hand mated and randomly assigned to 1 of 4 groups of 4 gilts each. Treated gilts had 108 mg of purified zearalenone added to their diet on postmating days (PMD) 2 to 6, 7 to 10, or 11 to 15. Control gilts were given the same diet without added zearalenone. On PMD 6, 10, and 15, control gilts had venous cannulas placed in the jugular vein, and blood samples were taken at 20-minute intervals for 4 hours before feeding and 4 hours after feeding. Samples were collected from treated gilts on the last day that zearalenone was consumed. Samples were analyzed for follicle stimulating hormone, luteinizing hormone (LH), and prolactin. Single blood samples were taken by venipuncture on PMD 8, 12, 16, 20, 24, and 28 and at euthanasia and were analyzed for serum concentration of progesterone and estradiol-17 beta. All gilts were euthanatized 30 to 32 days after mating, and fetal development was assessed. Three gilts that were given zearalenone on PMD 7 to 10 were not pregnant and had regressing corpora lutea on the ovaries at euthanasia. All other treated and control gilts were pregnant. Serum samples from treated gilts on PMD 10 and 15 had lower mean prolactin concentrations than did those from controls. The number of LH spikes were fewer (P less than 0.05) in gilts that were given zearalenone on PMD 15 compared with those in controls on PMD 15. Serum progesterone concentrations indicated that corpora lutea regressed between PMD 20 and 28 in nonpregnant gilts.(ABSTRACT TRUNCATED AT 250 WORDS)     

Uterine and fetal dynamics during early pregnancy in mares

Fetal activity and mobility and changes in diameter of the allantoic fluid compartment in the uterine horns were studied in mares between days 69 and 81 of pregnancy by use of transrectal ultrasonography (n = 12) and transcervical videoendoscopy (n = 8). The insertion tube of the videoendoscope was positioned within the allantoic sac to permit viewing of the fetus and entrance to each uterine horn. Each uterine horn was divided ultrasonographically into 3 segments of equal length, and the horns were designated on the basis of side of umbilical attachment (cord vs noncord horns). The diameter of the allantoic fluid compartment in the cornual segments increased (P less than 0.05) over the cranial (18.6 +/- 1.9 mm), middle (35.6 +/- 2.9 mm), and caudal (51.7 +/- 4.4 mm) segments, but differences between cord and noncord horns were not evident. Dynamic changes in diameter of the allantoic fluid compartment in cornual segments (ultrasonography) and at the entrance to each uterine horn (videoendoscopy) were detected (no significant difference between methods). During continuous videoendoscopic viewing (17 to 60 min/mare), extreme changes in allantoic fluid compartment diameter (76 to 100% of maximum to 0 to 25% of maximum or vice-versa) occurred an equivalent of 2.6 times/h/horn entrance; changes had an average duration of 3.4 minutes. A change from 100% (maximal diameter) to 0% (no visible lumen) or vice-versa occurred an equivalent of 1.3 times/h/horn entrance. Sometimes the uterine wall was so closely constricted++ around the fetal-amniotic unit that no intervening allantoic fluid was ultrasonographically detectable whereas at other times the uterus in the same location was widely dilated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Aspects of the antiluteolytic activity of the conceptus during early pregnancy in ewes

Experiments were undertaken to determine whether the conceptus renders a corpus luteum resistant to the luteolytic action of prostaglandin F2 alpha (PGF2 alpha), and modulates release of this prostaglandin by the uterus of early pregnant ewes. Prostaglandin F2 alpha was luteolytic when administered to indomethacin-treated ewes on d 10 and 11 of the estrous cycle. The same PGF2 alpha treatment was not luteolytic when applied on d 19 and 20 of pregnancy in ewes treated with indomethacin. Pulsatile release of PGF2 alpha (measured by 15-keto-13,14-dihydro PGF2 alpha-PGF2 alpha plasma level, PGFM) was observed between d 14 and 16 of the cycle but not during the same period of pregnancy. Ablation of the conceptus on d 17 resulted in progressive restoration of PGFM surges and subsequent luteolysis. Estradiol-17 beta (E2-17 beta) administration on d 12 of the cycle induced earlier PGFM surges and luteal regression. The same E2-17 beta treatment administered on d 14, 19 and 33 of pregnancy failed to induced PGFM pulses and luteolysis. In the absence of the conceptus (surgical ablation), E2-17 beta treatment was luteolytic (PGFM surges) on d 17 but not on d 33. We conclude that the conceptus controls the amount and pattern of PGF2 alpha released by the uterus, as well as the sensitivity of the uterus to E2-17 beta as early as d 14 of pregnancy. Simultaneously, an embryonic protective effect takes place at the luteal level.     

Expression analyses of sex steroid-regulated genes in neonatal rat hypothalamus

Estrogen plays an important role in sexual differentiation of the brain in rats during the perinatal period. To elucidate molecular mechanisms underlying sexual differentiation of the brain, in this study we investigated genes differentially expressed between sexes or induced to express by estrogen in neonatal rat hypothalamus using DNA microarray analysis in combination with real-time RT-PCR. It was found that the levels of expression of the genes encoding glutamic acid decarboxylase 65 and coronin 1b were higher in male than female hypothalamus on postnatal day (PN) 5 and those of collagen type 3 alpha1 and thioredoxin reductase 2 genes in female hypothalamus on PN5 were decreased and increased, respectively, by treatment with estradiol on PN2. Then the developmental changes in the expression of these 4 genes were examined from 1 day before the parturition to PN9, and they all showed sexual dimorphic patterns. In addition, dependence of the expression of these genes on either estradiol, testosterone or dihydrotestosterone during the neonatal period was confirmed. These results suggest that these four genes are involved in sexual differentiation of the rat brain, and that androgen per se as well as estrogen may take part in the processes.