-
Biology of Reproduction Feb 2014Embryo implantation and development requires the endometrial stromal cells (ESCs) to undergo decidualization. This differentiation process requires glucose utilization,...
Embryo implantation and development requires the endometrial stromal cells (ESCs) to undergo decidualization. This differentiation process requires glucose utilization, and blockade of the pentose phosphate pathway inhibits decidualization of ESCs both in vitro and in vivo. Glucose and fatty acids are energy substrates for many cell types, and fatty acid beta-oxidation is critical for embryo implantation. Here, we investigated whether beta-oxidation is required for decidualization of ESCs. As assessed by marker gene expression, decidualization of human primary ESCs was blocked by reducing activity of carnitine calmitoyltransferase I, the rate-limiting enzyme in beta-oxidation, either by short hairpin RNA-mediated silencing or by treatment with the inhibitor etomoxir. Ranolazine (RAN), a partial beta-oxidation inhibitor, blocked early decidualization of a human ESC line. However, decidualization resumed after several days, most likely due to a compensatory up-regulation of GLUT1 expression and an increase in glucose metabolism. Simultaneous inhibition of the beta-oxidation pathway with RAN and the pentose phosphate pathway with glucosamine (GlcN) impaired in vitro decidualization of human ESCs more strongly than inhibition of either pathway alone. These findings were confirmed in murine ESCs in vitro, and exposure to RAN plus GlcN inhibited decidualization in vivo in a deciduoma model. Finally, intrauterine implantation of time-release RAN and GlcN pellets reduced pup number. Importantly, pup number returned to normal after the end of the pellet-active period. This work indicates that both fatty acids and glucose metabolism pathways are important for ESC decidualization, and suggests novel pathways to target for the design of future nonhormonal contraceptives.
Topics: Animals; Cells, Cultured; Decidua; Endometrium; Fatty Acids; Female; Humans; Lipid Metabolism; Male; Metabolic Networks and Pathways; Mice; Mice, Inbred ICR; Oxidation-Reduction; Stromal Cells
PubMed: 24403548
DOI: 10.1095/biolreprod.113.113217 -
Journal of Toxicologic Pathology Mar 2013Uterine deciduomas were found in two female virgin rats, a 15-week-old Lewis rat and a 7-week-old Sprague-Dawley rat. The firm white nodules were located at the base of...
Uterine deciduomas were found in two female virgin rats, a 15-week-old Lewis rat and a 7-week-old Sprague-Dawley rat. The firm white nodules were located at the base of unilateral uterine horns and were approximately 6 mm and 4 mm in diameter. Histopathologically, the nodules were composed of three areas, each with a distinct type of proliferating cells: large epithelioid decidual cells with round nuclei, prominent nucleoli and abundant eosinophilic cytoplasm (antimesometrial region); compact spindle-shaped cells with oval nuclei and vacuolar cytoplasm (transitional region); and pleomorphic and spiny cells with round to oval nuclei and compact eosinophilic cytoplasm (mesometrial region). These cells proliferated in sheet-like arrangements and transformed into the other types of cells located in surrounding regions. Immunohistochemically, proliferating cells in all regions were strongly positive for proliferating cell nuclear antigen. The proliferating cells were positive for vimentin, and large decidual cells were positive for common acute lymphoblastic leukemia antigen 10, a marker of uterine interstitial cells. Large decidual cells were positive for α-smooth muscle actin and desmin, suggesting differentiation into muscular cells. Progesterone receptor was expressed in all cell types; however, estrogen receptor α was not expressed in the antimesometrial region. These extremely rare tumor-like nodules represent nonneoplastic lesions referred as decidual reactions of endometrial interstitial cells, and their biological behavior is that of a space-occupying benign tumor in young rats. Our cases might provide information as a historical control in toxicity and pharmacological studies in rats.
PubMed: 23723570
DOI: 10.1293/tox.26.61 -
Biology of Reproduction Jul 2013Embryo implantation in the uterus depends on decidualization of the endometrial stromal cells (ESCs), and glucose utilization via the pentose phosphate pathway is...
Embryo implantation in the uterus depends on decidualization of the endometrial stromal cells (ESCs), and glucose utilization via the pentose phosphate pathway is critical in this process. We hypothesized that the amino sugar glucosamine may block the pentose phosphate pathway via inhibition of the rate-limiting enzyme glucose-6-phosphate dehydrogenase in ESCs and therefore impair decidualization and embryo implantation, thus preventing pregnancy. Both human primary and immortalized ESCs were decidualized in vitro in the presence of 0, 2.5, or 5 mM glucosamine for 9 days. Viability assays demonstrated that glucosamine was well tolerated by human ESCs. Exposure of human ESCs to glucosamine resulted in significant decreases in the activity and expression of glucose-6-phosphate dehydrogenase and in the mRNA expression of the decidual markers prolactin, somatostatin, interleukin-15, and left-right determination factor 2. In mouse ESCs, expression of the decidual marker Prp decreased upon addition of glucosamine. In comparison with control mice, glucosamine-treated mice showed weak artificial deciduoma formation along the stimulated uterine horn. In a complementary in vivo experiment, a 60-day-release glucosamine (15, 150, or 1500 μg) or placebo pellet was implanted in a single uterine horn of mice. Mice with a glucosamine pellet delivered fewer live pups per litter than those with a control pellet, and pup number returned to normal after the end of the pellet-active period. In conclusion, glucosamine is a nonhormonal inhibitor of decidualization of both human and mouse ESCs and of pregnancy in mice. Our data indicate the potential for development of glucosamine as a novel, reversible, nonhormonal contraceptive.
Topics: Animals; Contraceptive Agents; Endometrium; Female; Glucosamine; Glucosephosphate Dehydrogenase; Humans; Litter Size; Mice; Pregnancy; Stromal Cells
PubMed: 23718985
DOI: 10.1095/biolreprod.113.108571 -
Journal of Cellular Biochemistry Oct 2012Transforming growth factor (TGF)-β and activin, members of TGF-β superfamily, are abundantly expressed in the endometrium and regulate decidualization of endometrial...
Transforming growth factor (TGF)-β and activin, members of TGF-β superfamily, are abundantly expressed in the endometrium and regulate decidualization of endometrial stroma. Smad2 and Smad3 are receptor-regulated Smads (R-Smads) that transduce extracellular TGF-β/activin/Nodal signaling. In situ hybridization results showed that Smad3 was highly expressed in the decidual zone during the peri-implantation period in mice. By using artificial decidualization, we found that Smad3 null mice showed partially compromised decidualization. We therefore hypothesized that Smad2 might compensate for the function of Smad3 during the process of decidualization. Smad2 was also highly expressed in the decidual zone and phosphorylated Smad2 was much more abundantly increased in the deciduoma of Smad3 null mice than for wild-type (WT) mice. We further employed an in vitro uterine stromal cell decidualization model, and found that decidual prolactin-related protein (dPRP) and cyclin D3, which are well-known markers for decidual cells, were significantly down-regulated in Smad3 null decidual cells, and were much more significantly reduced when the expression of Smad2 was simultaneously silenced by its siRNA (P < 0.05). However, the expression levels of dPRP and cyclin D3 remained the same when Smad2 was silenced in WT decidual cells. Collectively, these findings provide evidence for an important role of Smad3 in decidualization and suggest that Smad2 and Smad3 may have redundant roles in decidualization.
Topics: Animals; Cyclin D3; Decidua; Dinoprostone; Embryo Implantation; Embryo, Mammalian; Female; Gene Expression Regulation, Developmental; Heterozygote; Homozygote; Male; Mice; Mice, Knockout; Ovariectomy; Phosphorylation; Pregnancy; Primary Cell Culture; RNA, Small Interfering; Smad2 Protein; Smad3 Protein; Stromal Cells; Transforming Growth Factor beta
PubMed: 22644778
DOI: 10.1002/jcb.24204 -
The Journal of Biological Chemistry May 2012Embryo implantation into the maternal uterus is a crucial step for the successful establishment of mammalian pregnancy. Following the attachment of embryo to the uterine...
Progesterone and DNA damage encourage uterine cell proliferation and decidualization through up-regulating ribonucleotide reductase 2 expression during early pregnancy in mice.
Embryo implantation into the maternal uterus is a crucial step for the successful establishment of mammalian pregnancy. Following the attachment of embryo to the uterine luminal epithelium, uterine stromal cells undergo steroid hormone-dependent decidualization, which is characterized by stromal cell proliferation and differentiation. The mechanisms underlying steroid hormone-induced stromal cell proliferation and differentiation during decidualization are still poorly understood. Ribonucleotide reductase, consisting of two subunits (RRM1 and RRM2), is a rate-limiting enzyme in deoxynucleotide production for DNA synthesis and plays an important role in cell proliferation and tumorgenicity. Based on our microarray analysis, Rrm2 expression was significantly higher at implantation sites compared with interimplantation sites in mouse uterus. However, the expression, regulation, and function of RRM2 in mouse uterus during embryo implantation and decidualization are still unknown. Here we show that although both RRM1 and RRM2 expression are markedly induced in mouse uterine stromal cells undergoing decidualization, only RRM2 is regulated by progesterone, a key regulator of decidualization. Further studies showed that the induction of progesterone on RRM2 expression in stromal cells is mediated by the AKT/c-MYC pathway. RRM2 can also be induced by replication stress and DNA damage during decidualization through the ATR/ATM-CHK1-E2F1 pathway. The weight of implantation sites and deciduoma was effectively reduced by specific inhibitors for RRM2. The expression of decidual/trophoblast prolactin-related protein (Dtprp), a reliable marker for decidualization in mice, was significantly reduced in deciduoma and steroid-induced decidual cells after HU treatment. Therefore, RRM2 may be an important effector of progesterone signaling to induce cell proliferation and decidualization in mouse uterus.
Topics: Animals; Blotting, Western; Cell Proliferation; Cells, Cultured; DNA Damage; Decidua; E2F1 Transcription Factor; Embryo Implantation; Female; Gene Expression Regulation, Developmental; Hydroxyurea; Male; Mice; Ovariectomy; Pregnancy; Progesterone; Protein Binding; Proto-Oncogene Proteins c-myc; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleoside Diphosphate Reductase; Ribonucleotide Reductases; Signal Transduction; Stromal Cells; Time Factors; Uterus
PubMed: 22403396
DOI: 10.1074/jbc.M111.308023 -
FASEB Journal : Official Publication of... Jan 2012Uterine receptivity implies a dialogue between the hormonally primed maternal endometrium and the free-floating blastocyst. Endometrial stromal cells proliferate, avert...
Uterine receptivity implies a dialogue between the hormonally primed maternal endometrium and the free-floating blastocyst. Endometrial stromal cells proliferate, avert apoptosis, and undergo decidualization in preparation for implantation; however, the molecular mechanisms that underlie differentiation into the decidual phenotype remain largely undefined. The Notch family of transmembrane receptors transduce extracellular signals responsible for cell survival, cell-to-cell communication, and differentiation, all fundamental processes for decidualization and pregnancy. Using a murine artificial decidualization model, pharmacological inhibition of Notch signaling by γ-secretase inhibition resulted in a significantly decreased deciduoma. Furthermore, a progesterone receptor (PR)-Cre Notch1 bigenic (Notch1(d/d)) confirmed a Notch1-dependent hypomorphic decidual phenotype. Microarray and pathway analysis, following Notch1 ablation, demonstrated significantly altered signaling repertoire. Concomitantly, hierarchical clustering demonstrated Notch1-dependent differences in gene expression. Uteri deprived of Notch1 signaling demonstrated decreased cellular proliferation; namely, reduced proliferation-specific antigen, Ki67, altered p21, cdk6, and cyclinD activity and an increased apoptotic-profile, cleaved caspase-3, Bad, and attenuated Bcl2. The results demonstrate that the preimplantation uterus relies on Notch signaling to inhibit apoptosis of stromal fibroblasts and regulate cell cycle progression, which together promotes successful decidualization. In summary, Notch1 signaling modulates multiple signaling mechanisms crucial for decidualization and these studies provide additional perspectives to the coordination of multiple signaling modalities required during decidualization.
Topics: Animals; Apoptosis; Cell Communication; Cell Differentiation; Cell Division; Cytoskeleton; Decidua; Embryo Implantation; Female; Mice; Mice, Inbred C57BL; Mice, Knockout; Oligonucleotide Array Sequence Analysis; Ovariectomy; Pregnancy; Pregnancy, Animal; Receptor, Notch1; Signal Transduction; Stromal Cells
PubMed: 21990372
DOI: 10.1096/fj.11-184663 -
Fertility and Sterility Jun 2011Fatty acid-binding protein 4 (Fabp4) is highly expressed in the secondary decidual zone of mouse decidua and deciduoma and stromal cells under in vitro decidualization....
Fatty acid-binding protein 4 (Fabp4) is highly expressed in the secondary decidual zone of mouse decidua and deciduoma and stromal cells under in vitro decidualization. Dtprp, a well-known marker of in vitro decidualization, is diminished by small interfering RNA against Fabp4 and FABP4 inhibitor and stimulated through Fabp4 overexpression.
Topics: Animals; Biphenyl Compounds; Cells, Cultured; Decidua; Deciduoma; Embryo Implantation; Estradiol; Fatty Acid-Binding Proteins; Female; Gene Expression Regulation; Humans; Immunohistochemistry; In Situ Hybridization; Mice; Pregnancy; Progesterone; Prolactin; Pyrazoles; RNA Interference; RNA, Messenger; Stromal Cells; Time Factors; Transfection
PubMed: 21704217
DOI: 10.1016/j.fertnstert.2011.05.052 -
Cell and Tissue Research Jun 2011Tumor necrosis factor receptor subfamily 9 (TNFRSF9) plays a potentially important general role in immune function. Tnfrsf9 gene expression has previously been...
Tumor necrosis factor receptor subfamily 9 (TNFRSF9) plays a potentially important general role in immune function. Tnfrsf9 gene expression has previously been characterized in late pregnant mouse uterus and placenta. However, little is known about its expression in the uterus during the implantation phase of early pregnancy. We have assessed the levels and localization of Tnfrsf9 expression in the mouse uterus and conceptus during implantation. Relative Tnfrsf9 mRNA levels were significantly higher in implantation than in non-implantation site tissue on days 6.5-8.5 of pregnancy. This increase did not depend on the presence of the conceptus, as mRNA levels were not significantly different between pregnant implantation sites and artificially induced deciduomas. Localization by in situ hybridization revealed a subpopulation of endothelial and uterine natural killer cells expressing Tnfrsf9 in the endometrium during implantation. In the developing conceptus, primary trophoblast giant and ectoplacental cells expressed Tnfrsf9 on days 6.5-8.5, followed by expression in the trophoblast giant cell layers surrounding the conceptus on day 9.5 of pregnancy. Two main splice forms of Tnfrsf9 mRNA exist and encode proteins with distinct biological functions; both mRNA splice forms were present in uterine and conceptus tissues as determined by reverse transcription with the polymerase chain reaction. Thus, both membrane and soluble forms of Tnfrsf9 are expressed in specific cell types of the uterus and conceptus during the progression of implantation in mice and possibly have an important function in this process.
Topics: Animals; Decidua; Embryo Implantation; Female; Gene Expression; Male; Mice; Mice, Knockout; Pregnancy; RNA, Messenger; Tumor Necrosis Factor Receptor Superfamily, Member 9; Uterus
PubMed: 21560035
DOI: 10.1007/s00441-011-1171-0 -
Reproduction (Cambridge, England) Apr 2011During pregnancy in several species including humans and rodents, the endometrium undergoes decidualization. This process of differentiation from endometrial to decidual...
During pregnancy in several species including humans and rodents, the endometrium undergoes decidualization. This process of differentiation from endometrial to decidual tissue occurs only after the onset of implantation in mice. It can also be artificially induced causing the formation of deciduomal tissue. The purpose of this study was to compare the gene expression profile of the developing decidua in pregnant mice with the deciduoma formed after artificial induction in an effort to identify conceptus-influenced changes in uterine gene expression during decidualization. We induced decidualization artificially by transferring blastocyst-sized ConA-coated agarose beads into the uterus on day 2.5 of pseudopregnancy. Recently published work has found this model to be more 'physiological' than other methods. Total RNA was isolated from blastocyst and bead-induced 'implantation' sites of the uteri of day 7.5 pregnant (decidua) and pseudopregnant (deciduoma) mice respectively. This RNA was then used for microarray analysis using Mouse Illumina BeadArray chips. This analysis revealed potential differential mRNA levels of only 45 genes between the decidua and bead-induced deciduoma tissues. We confirmed the differential mRNA levels of 31 of these genes using quantitative RT-PCR. Finally, the level and localization of some of the mRNAs for select genes (Aldh3a1, Bcmo1, Guca2b, and Inhbb) identified by our microarray analysis were examined in more detail. This study provides the identity of a small set of genes whose expression in the uterus during decidualization may be influenced by molecular signals from the conceptus.
Topics: Animals; Blastocyst; Cluster Analysis; Decidua; Female; Gene Expression; Gene Expression Profiling; Male; Mice; Microarray Analysis; Placentation; Pregnancy; Uterus
PubMed: 21300692
DOI: 10.1530/REP-10-0358 -
Pharmacognosy Research May 2010Women experience menopause differently across the world, in terms of their symptomology. Many experience symptoms of menopause like hot flashes, joint pain and loss of...
Women experience menopause differently across the world, in terms of their symptomology. Many experience symptoms of menopause like hot flashes, joint pain and loss of libido. Estrogen replacement is the prescribed therapy for most of the sexual dysfunction observed in menopausal women. Many women are reluctant to use exogenous hormone therapy for treatment of menopausal symptoms and are turning to botanical and dietary supplements for relief. In the present study IND-HE (friedelin rich fraction) was studied for estrogenic activity as well as its effect on sexual behavior in overiectomized female Wistar rats.The rats were divided into 4 groups of six rats each. The Group 1 received distilled water, Group II - IND-HE (75 mg/kg p. o.), Group III - IND-HE (100 mg/kg p. o.) and Group IV received estrogen (estradiol) (1 mg/kg in olive oil suspension, s.c. bi-weekly). The treatment period was 8 weeks. On 1 day, one month and two month of treatment the sexual behavior was studied. At the end of the treatment the blood was withdrawn from retro-orbital plexus. The animals were sacrificed and uterus was removed, weighed and histology was studied. In different group of rats estrous cycle was studied which indicate estrogenic activity and for progestogenic activity of deciduoma formation was studied.The result indicated that IND-HE (75 and 100 mg/kg p.o.) improved sexual behavior parameters. IND-HE (75 and 100) significantly (P< 0.01) decreased darting and hopping latency. The darting frequency and hopping frequency was significantly (P< 0.01) improved in IND-HE (75 and100 mg/kg p.o.) as well as estrogen group. Lordosis interval (LI) was increased significantly in estrogen group after 1(st) month (P< 0.05), and after 2(nd) month (P< 0.01). IND-HE (100) treatment showed increase in LI after 1(st) month (P< 0.05) remained during 2(nd) month (P< 0.01). While IND-HE (75) treatment increased LI only after 2(nd) month (P< 0.05).IND-HE (75 and 100 mg/kg p.o.) showed estrogenic activity as indicated by vaginal cornification, increase in uterine weight and rise in serum estrogen.
PubMed: 21808556
DOI: 10.4103/0974-8490.65507