Placental Trophoblast Giant Cells Research Articles

Adhesion G protein-coupled receptor Gpr126/Adgrg6 is essential for placental development.

Mutations in the G protein–coupled receptor GPR126/ADGRG6 cause human diseases, including defective peripheral nervous system (PNS) myelination. To study GPR126 function, we generated new genetic mice and zebrafish models. Murine Gpr126 is expressed in developing heart endocardium, and global Gpr126 inactivation is embryonically lethal, with mutants having thin-walled ventricles but unaffected heart patterning or maturation. Endocardial-specific Gpr126 deletion does not affect heart development or function, and transgenic endocardial GPR126 expression fails to rescue lethality in Gpr126-null mice. Zebrafish gpr126 mutants display unaffected heart development. Gpr126 is also expressed in placental trophoblast giant cells. Gpr126-null mice with a heterozygous placenta survive but exhibit GPR126-defective PNS phenotype. In contrast, Gpr126-null embryos with homozygous mutant placenta die but are rescued by placental GPR126 expression. Gpr126-deficient placentas display down-regulation of preeclampsia markers Mmp9, Cts7, and Cts8. We propose that the placenta-heart axis accounts for heart abnormalities secondary to placental defects in Gpr126 mutants.

More of a good thing or less of a bad thing: gene copy number variation in polyploid cells of the placenta.

More of a good thing or less of a bad thing: gene copy number variation in polyploid cells of the placenta.

A pregnant mouse model for the vertical transmission of Brucella melitensis

Abortion is the major clinical sign of brucellosis in animals but little is known about the underlying mechanisms. This study was designed to evaluate a pregnant mouse model for the vertical transmission of Brucella melitensis using four infectious doses: 103 colony-forming units (CFU), 104 CFU, 105 CFU, and 106 CFU. During the experimental period, no instances of abortion were recorded, but stillbirths were observed in the groups infected with doses of 104 CFU and higher.Regardless of whether the fetuses were stillborn or alive, transmission of bacteria to the fetus and bacterial replication in the cytoplasm of placental trophoblast giant cells were detected. A higher degree of bacterial colonization was found in the placenta than in the spleen or fetus. Doses of 105 CFU of B. melitensis or higher produced a severe, necrotizing placentitis similar to the pathological damage observed in ruminants. The data suggest that experimental murine brucellosis resembles ruminant brucellosis and represents a potential model for studying the pathogenic mechanisms of B. melitensis.

Mannose receptor, C type 1 contributes to bacterial uptake by placental trophoblast giant cells

During pregnancy, maternal immune function is strictly controlled and immune tolerance is induced. Trophoblast giant (TG) cells exhibit phagocytic activity and show macrophage-like activity against microorganisms in the placenta. However, details of molecular receptors and mechanisms for uptake by TG cells have not been clarified. In this study, we investigated the involvement of the mannose receptor, C type 1 (MRC1), in the uptake of the abortion-inducible bacterium Listeria monocytogenes and abortion-uninducible bacteria Bacillus subtilis and Escherichia coli by TG cells differentiated from a mouse trophoblast stem cell line in vitro. Knockdown of MRC1 inhibited the uptake of all of these bacteria, as did the blocking of MRC1 by MRC1 ligands. The uptake of bacteria by MRC1 delayed the maturation of phagolysosomes. These findings suggest that MRC1 plays an important role in the uptake of various bacteria by TG cells and may provide an opportunity for those bacteria to escape from phagosomes.

Transcription of Steroidogenic Acute Regulatory Protein in the Rodent Ovary and Placenta: Alternative Modes of Cyclic Adenosine 3′, 5′-Monophosphate Dependent and Independent Regulation

Steroid hormone synthesis is a vital function of the adrenal cortex, serves a critical role in gonadal function, and maintains pregnancy if normally executed in the placenta. The substrate for the synthesis of all steroid hormones is cholesterol, and its conversion to the first steroid, pregnenolone, by the cholesterol side-chain cleavage cytochrome P450 (CYP11A1) enzyme complex takes place in the inner mitochondrial membranes. Steroidogenic acute regulatory protein (STAR) facilitates the rate-limiting transfer of cholesterol from the outer mitochondrial membrane to CYP11A1 located in the inner organelle membranes. The current study explored the mechanisms controlling transcription of the Star gene in primary cell cultures of mouse placental trophoblast giant cells and rat ovarian granulosa cells examined throughout the course of their functional differentiation. Our findings show that the cis-elements required for Star transcription in the rodent placenta and the ovary are centered in a relatively small proximal region of the promoter. In placental trophoblast giant cells, cAMP is required for activation of the Star promoter, and the cis-elements mediating a maximal response were defined as cAMP response element 2 and GATA. EMSA studies show that placental cAMP-responsive element binding protein (CREB)-1 and activating transcription factor-2 (ATF2) bind to a -81/-78 sequence, whereas GATA-2 binds to a -66/-61 sequence. In comparison, patterns of Star regulation in the ovary suggested tissue-specific and developmental controlled modes of Star transcription. During the follicular phase, FSH/cAMP induced CREB-1 dependent activity, whereas upon luteinization STAR expression becomes cAMP and CREB independent, a functional shift conferred by FOS-related antigen-2 displacement of CREB-1 binding, and the appearance of a new requirement for CCAAT enhancer-binding protein beta and steroidogenic factor 1 that bind to upstream elements (-117/-95). These findings suggest that during evolution, the promoters of the Star gene acquired nonconsensus sequence elements enabling expression of a single gene in different organs, or allowing dynamic temporal changes corresponding to progressing phases of differentiation in a given cell type.

Differentiation of Placental Trophoblast Giant Cells Requires Downregulation of p53 and Rb

Trophoblast giant cells in the rodent placenta form the outermost layer of the extraembryonic compartment, establish direct contact with maternal cells, and produce a number of pregnancy-specific cytokine hormones. Giant cells differentiate from proliferative trophoblasts as they exit the cell cycle and enter a genome-amplifying endocycle, a process we show involves decreased expression of the G1 checkpoint proteins p53 and Rb. Although p53 mRNA levels are unchanged in proliferative compared to differentiated trophoblasts, p53 protein levels are markedly reduced in giant cells. Forced expression of wild type p53 in trophoblasts inhibits differentiation, and expression of a dominant negative p53 peptide stimulates differentiation. Consistent with the loss of p53 protein, differentiated trophoblasts become resistant to apoptosis-inducing agents. Decreased expression of Rb is also detected during differentiation, and overexpression of Rb in trophoblasts inhibits giant cell differentiation. Although an increase in E2F activity would be expected with the loss of Rb, what is observed is an overall decrease in E2F DNA-binding complexes, a shift to new complexes, and a decrease in E2F-dependent gene expression in differentiating trophoblasts. Overall, these results indicate that the combination of a decrease in p53 and Rb represents a functionally important part of the transition of trophoblasts from a proliferative cell cycle to an endocycle in the giant cell differentiation programme.

Cyclins E1 and E2 are required for endoreplication in placental trophoblast giant cells.

In mammalian cells, cyclin E-CDK2 complexes are activated in the late G1 phase of the cell cycle and are believed to have an essential role in promoting S-phase entry. We have targeted the murine genes CCNE1 and CCNE2, encoding cyclins E1 and E2. Whereas single knockout mice were viable, double knockout embryos died around midgestation. Strikingly, however, these embryos showed no overt defects in cell proliferation. Instead, we observed developmental phenotypes consistent with placental dysfunction. Mutant placentas had an overall normal structure, but the nuclei of trophoblast giant cells, which normally undergo endoreplication and reach elevated ploidies, showed a marked reduction in DNA content. We derived trophoblast stem cells from double knockout E3.5 blastocysts. These cells retained the ability to differentiate into giant cells in vitro, but were unable to undergo multiple rounds of DNA synthesis, demonstrating that the lack of endoreplication was a cell-autonomous defect. Thus, during embryonic development, the needs for E-type cyclins can be overcome in mitotic cycles but not in endoreplicating cells.

The effects of morphine on cell proliferation.

There is increasing evidence that endogenous opioid peptides ("enkephalins") and other neurotransmitters have widespread, receptor-mediated roles as growth regulators in non-neuronal cells and tissues. For example, it is now believed that enkephalins produced in placental trophoblast giant cells have multiple roles in supporting embryo growth, and in maternal adaptation to pregnancy. Since plant and synthetic narcotics (e.g., morphine) bind to the same receptors, the questions immediately arise: Do narcotics also have actions as growth regulators? If so, do these actions have physiological significance in addicts? Recent work on the first of these questions is covered in this review. While the greatest volume of research has been focused on the proliferative effects of narcotics for cells of the immune system, the roles of opioid peptides and narcotics on the growth of a variety of other cells has come under study recently.

Identification of trophoblast-specific regulatory elements in the mouse placental lactogen II gene.

Placental lactogen II, the major ligand for the PRL receptor during the second half of gestation in rodents, is synthesized specifically by placental trophoblast giant cells. A transient transgenic analysis has been used to localize the giant cell-specific regulatory region within the mouse placental lactogen II gene to sequences between -1340 and -2019 upstream of the transcriptional start site. More precise mapping of the regulatory elements has been accomplished by transfection of promoter constructs into Rcho-1 trophoblast cells, resulting in the characterization of two positive regulatory elements in the -1471 to -1340 region; two other regulatory elements have been implicated but not further characterized, a negative regulatory element between -2019 and -1778 and another positive element within the region from -1340 to -569. Both of the characterized positive regulatory elements are recognized by factors that are enriched in differentiated giant cells compared with proliferative trophoblasts, and these factors are either absent or at low levels in fibroblasts. The complexes that form on the two elements are distinct and neither element competes with the other for factor binding, thus implicating at least two different regulatory elements in late-gestational trophoblast giant cell-specific gene expression.

Placental-specific expression from the mouse placental lactogen II gene promoter.

The gene for mouse placental lactogen II (mPL-II) has been isolated and characterized. This gene contains five exons, with a transcription start site 59 nucleotides upstream of the translation initiation ATG. Introduction of a DNA construct containing 2.7 kilobases of sequence upstream of the mPL-II transcription initiation site directed the synthesis of a linked coding region for the simian virus 40 large and small tumor antigens in placental trophoblast giant cells of transgenic mice. The pattern of simian virus 40 transgene expression in the placenta was indistinguishable from that of the endogenous mPL-II gene. In contrast, the first 569 base pairs upstream of the transcription start site proved insufficient to direct placental expression. Thus, one or more elements required for placental trophoblast giant cell expression have been localized to a region between -2700 and -569 of the mPL-II gene.