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Indirect immunofluorescent localization of prolactin to the cytoplasm of decidua and trophoblast cells in human placental membranes at term.

An indirect immunofluorescent technique was used to determine the localization of cytoplasmic human PRL (hPRL) in fresh and incubated human placental membranes at term. In both fresh and 8-h incubated samples of amnion, amniochorion decidua, or chorion decidua obtained from three placentas, we found specific reproducible localization of hPRL to the cytoplasm of decidua and trophoblast cells. The decidua cells appeared to be the most intensely fluorescent. No specific hPRL immunofluorescence was noted in the amniotic epithelium of fresh or incubated samples of amnion and amniochorion decidua. These data suggest that the trophoblast decidua cell layer is the site of PRL localization and possibly synthesis in placental membranes at term and may be the origin of amniotic fluid PRL in humans.

Cytoplasm

Is human decidua a specialized endocrine organ?

The production of protein hormones by human placenta, amnion, chorion, and decidua capsularis was studied in in vitro experiments to establish whether the decidua could be considered a specialized structure for the release of PRL. The tissues were incubated in the incubation medium, we found a highly significant rise of PRL during culture of the decidua, while no increase was noticed during culture of the placenta or amnion. Conversely, chorionic somatomammotropin and CG increased greatly and quickly during culture of placenta but not during culture of other tissues. No significant change was found in GH. The total PRL released into the medium from decidua was 3 times higher than the initial PRL content of this sittue; addition of puromycin to the incubation medium reduced both the tissue content and the release of PRL to almost 50% of the control values. This result raises the possibility of a specific endocrine activity of decidua capsularis. The PRL-secreint cells of the decidua probably do not possess dopamine receptors, since bromocriptine, when added to the medium, did not influence the release of PRL, confirming our previous in vivo observations.

Amnion

Synthesis of prolactin by human decidua in vitro.

To determine whether human decidua and/or chorion synthesizes and secretes prolactin, explants of decidua obtained at Caesarian section and explants of chorion from the membranes separating dizygotic twins were cultured for periods of up to 6 days. The decidual explants released 366 +/- 37 ng prolactin/100 mg tissue (mean +/- S.D.) during each day in culture and incorporated 3H-labelled amino acids into immunoprecipitable prolactin. In the radioimmunoassay for prolactin, serial dilutions of incubation medium displaced 125I-labelled prolactin parallel to the displacement by pituitary prolactin and the prolactin in the medium eluted from Sephadex G-150 in a position indentical to that of pituitary prolactin. Chorionic explants released prolactin into the incubation medium during day 1 of culture only and did not incorporate 3H-labelled amino acids into prolactin. These results demonstrate that prolactin is synthesized by the decidua and not by the chorion and suggest that the decidua is the source of prolactin in amniotic fluid.

Amino Acids

Adenylate cyclase from human decidua parietalis.

Adenylate cyclase (EC 4.6.1.1) activity was demonstrated in membrane fractions of human decidua parietalis. Adenylate cyclase activity was associated with membrane fractions of decidua but not chorion or amnion. Prostaglandins PGE1 and PGE2 but neither PGD2 nor PGF2 alpha stimulated this activity at gestational ages 18 to 40 weeks. Fluoride and guanosine triphosphate stimulated decidual adenylate cyclase; however, epinephrine-stimulated adenylate cyclase activity was demonstrable only in the presence of guanosine triphosphate. The possible roles of adenylate cyclase activity in the decidua are discussed.

Adenylyl Cyclases

[Demonstration of immunoreactive GIF-like substance in villi and decidua by radioimmunoassay and immunofluorescence (author's transl)].

Since Arimura et al (1975) reported the radioimmunoassay for somatostatin (GIF), the concentration of GIF in various organ and brain regions were determined by radio-immunoassay. Dubois et al (1975) reported that immunohistochemically somatostatin was located in the discrete cells of the pancreas as well as the hypothalamus, and from this result, they presented the concept of local hormone instead of systemic hormone which was up to that time accepted in endocrinology. In this study, we developed the high specific anti-GIF serum using rabbits, and with the micro immunodiffusion method, we demonstrated that the precipitin band formed a circular fusion between the GIF and anti-GIF serum. This pattern of reaction was also seen in decidual immunodiffusion. In addition, we developed the radioimmunoassay for GIF using this anti-serum and measured immunoreactive GIF-like substances in villi and decidua of early pregnancy. The concentration of GIF-like substances with 2 N acetic acid extracted of villi and decidua were 0 to 30 pg/0.1 g dry weight. At the same time, we demonstrated the presence of GIF-like substance-containing cells in the villi and decidua by indirect immunofluorescent method. The intensity of immunofluorescence was in cytotrophoblast rather than syncytiotrophoblast, and decidual stromal cell also reacted to the immunofluorescence.

Chorionic Villi

Demonstration of immunoreactive somatostatin-like substance in villi and decidua in early pregnancy.

With the use of the radioimmunoassay for growth hormone--releasing inhibiting factor (GIF), it was found that measurable amounts of GIF-like substance existed in the chorionic villi and decidua of pregnant women. The indirect immunofluorescent method revealed that the higher intensity of GIF-like immunofluorescence was presented in cytotrophoblasts rather than in syncytiotrophoblasts of the villi and in stromal cells of the decidua.

Antigens

Human decidua and uterine contractility.

Physiological saline solution injected into the uterus by the extraamniotic route was found to damage the decidua and rupture its lysosomes. The mechanism by which this procedure provokes abortion may be through the release of lysosomal phospholipase A2 (EC 3.1.1.4), an enzyme thought to regulate the formation of polyunsaturated fatty acids utilized for prostaglandin synthesis. A similar sequence of events may be involved in the initiation of spontaneous labour, since decidual cells obtained at elective Caesarean section at term exhibited degenerative changes and signs of release of a lysosomal marker enzyme. The cause of this release is unknown.

Abortifacient Agents, Nonsteroidal

Work in progress. Occurence of phospholipase A1 and A2 in human decidua.

Phospholipase A2, an enzyme which may regulate the formation of polyunsaturated fatty acids utilized for prostaglandin synthesis, was found to have significant higher activity in decidual than in myometrial tissue. The major part of phospholipase A2 in the decidua had an acid pH optimum, which indicates that most of the enzyme is stored in the lysosomes of this tissue. These findings, together with previous observations, lend further support to the view that lysosomal phospholipase A2 released within decidual cells might be a trigger of abortion and parturition.

Buffers

Ultrastructural relationships between decidua, trophoblast and lymphocytes at the beginning of human pregnancy.

Between the 20th and 40th day of pregnancy, human decidual cells in the endometrial connective tissue come into direct contact with the trophoblastic cells eroding the stromal tissue. Fibrillar deposits of maternal origin were randomly distributed between the maternal and fetal sites and did not form a continuous barrier. Structural features were prominent in the maternal and embryonic cells. Large decidual cells were intermingled with trophoblastic cells, and there was little intercellular material. Small developing decidual cells arose around the maternal precapillary arterioles and capillaries and were closely accompanied by lymphocytes. Fully developed decidual cells exhibited a fine structure characteristic of a state of active secretion, and trophoblastic cells gave structural indications of producing protein. The aggregates of decidual and trophoblastic cells in the human endometrium during early pregnancy ('deciduotrophoblastic complex') may function as a local humoral regulator releasing chemical factors and partaking in the establishment of immunological privilege at implantation.

Animals

Intra-uterine tissues from late-pregnant rhesus monkeys (Macaca mulatta) produce 6-oxo-prostaglandin F1 alpha in vitro.

The rates of production of 6-oxo-prostaglandin F1 alpha (6-oxo-PGF1 alpha) in vitro by intra-uterine tissues taken from late-pregnant monkeys at Caesarean section have been determined. For tissues obtained between days 140 and 149 of pregnancy (late pregnancy) the general quantitative order of rates of production (per unit weight) was decidua basalis greater than placenta greater than decidua parietalis greater than amnion greater than chorion = myometrium. When tissues were taken between days 160 and 168 of pregnancy (near term) this order was placenta greater than decidua parietalis = amnion greater than myometrium = decidua basalis greater than chorion. There was a significant reduction near term in the rate of production of 6-oxo-PGF1 alpha by decidua basalis; all other tissues exhibited similar rates of production at the two gestational periods investigated.

Amnion

Specific change in the direction of prostaglandin synthesis by intra-uterine tissues of the rhesus monkey (Macaca mulatta) during late pregnancy.

The rates of production of prostaglandin E (PGE), prostaglandin F (PGF) and 13, 14-dihydro-15-oxo-prostaglandin F (PGFM) by intra-uterine tissues from pregnant monkeys in vitro have been determined using a method of tissue superfusion. The amnion, chorion, placenta, decidua basalis, decidua parietalis and myometrium were obtained at Caesarean section during late pregnancy. Production of PGE by all tissues was significantly lower at term than during late pregnancy, whereas production of PGF by the amnion, chorion, decidua parietalis and myometrium was significantly greater. All tissues produced significantly more PGE than PGF and also, excepting the decidua basalis and decidua parietalis, more PGFM than PGF. Close to parturition the amnion was quantitatively (per unit weight) the major source of prostaglandins. It is suggested that a specific change in the direction of prostaglandin synthesis by intra-uterine tissues occurs near parturition in the rhesus monkey.

Animals

[Studies on the activities of glycogen synthetase and glycogen phosphorylase in the human endometrium (author's transl)].

Glycogen content, glycogen synthetase and glycogen phosphorylase were studied in the endometrial tissues of 28 women with normal menstrual cycles and in the decidual tissues of 24 women with normal early pregnancies. The endometrial glycogen synthetase enzyme increased gradually from the proliferative phase to the secretory phase and reached maximal activity during the sixteenth to twenty-third days of the cycle, a time coincident with maximal glycogen content, while the glycogen phosphorylase reached its maximal activity on and after the twenty-fourth day of the cycle. In addition, glycogen phosphorylase activity in the decidual tissue during the early period of gestation (4--10 weeks) was lower than that in the endometrial tissue during the late secretory phase of the cycle, and in particular, the active form (-AMP) was significantly (p less than 0.005) low, with the result that the glycogen content in the decidua was significantly (p less than 0.05) higher than that in the endometrium. On the other hand, in 20 normal pregnant women during the 6--10th weeks of gestation, the glycogen contents in the decidua and in the placental villi were 599 +/- 44 mg/100 g wet weight (mean +/- standard error of the mean) and 731 +/- 55 mg/100 g, respectively, but the difference between them was statistically not significant. The levels of glycogen synthetase and glycogen phosphorylase enzyme in the decidua were significantly (p less than 0.005) higher than those in the placental villi.

Chorionic Villi

Therapeutic effect and anti-inflammatory mechanism of modified Shoutai pills against lipopolysaccharide-induced miscarriage in mice.

OBJECTIVE: To elucidate the anti-inflammatory mechanisms of modified Shoutai pills (, MSTP) in miscarriages, we performed transcriptome sequencing on the decidua and placental tissues of pregnancy mice. METHODS: The therapeutic effects and anti-inflammatory mechanisms of MSTP were studied in mice with lipopolysaccharide (LPS)-induced miscarriage. First, the effects of MSTP on pregnancy outcomes and the maternal-fetal interface, in LPS-induced miscarriage mice were examined. RNA sequencing was used to further investigate gene expression changes in LPS-induced miscarriage mice and to assess the effects of MSTP intervention. Finally, the expression levels of inflammation-related genes in the decidua and placental tissues were determined using quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: A high dose of MSTP significantly decreased the resorption rate (P < 0.05) and reduced apoptosis of the decidua and placental tissues in mice. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed that inflammatory and immune-related signals were enriched. qRT-PCR results confirmed that in decidual and placental tissues, MSTP reduced the gene expression levels of toll-like receptor 4 (TLR4), nuclear factor kappa-B (NF-&#x3ba;B), c-Jun N-terminal kinase 1, p38, and tumor necrosis factor-&#x3b1;. CONCLUSIONS: In this study, we demonstrated that MSTP effectively prevented embryo loss with an anti-inflammatory mechanism through downregulation of the TLR4-NF-&#x3ba;B/ MAPK signaling pathway, in LPS-induced miscarriage mice model. To our knowledge, this is the first study to reveal the therapeutic mechanism of MSTP in LPS-induced miscarriage in mice.

Animals

[Thymidylate synthetase in the antimesometrial and mesometrial portions of decidual tissue under normal conditions and following application of the antifolate preparation chloridine].

Specific activity of thymidylate synthetase (TMS) under normal conditions and after the action of pyrimethamine was investigated in antimesometrial (A) and mesometrial (M) parts of rat decidua. On the 9th-11th days of pregnancy specific activity of TMS in the A part proved to be higher than in the M part of decidua, and the former one reached its maximum by the 10th day. Specific activity of TMS in the M part decreased gradually from the 9th to 11th days of gestation. Pyrimethamine, applied on the 9th day of pregnancy caused a wave-like increase in the specific activity of TMS in the A part; this elevation was not so pronounced in the M part; A study of the physico-chemical properties of TMS from the decidua and the embryos of rats showed that the enzyme had a molecular weight of 58000, an optimal pH of 6.9, and could be quickly inactivated by heating.

Animals

Smooth muscle within ovarian decidual nodules: a link to leiomyomatosis peritonealis disseminata?

Evidence is presented for the coexistence of smooth muscle and decidual cells in nodules on and within the ovarian tunica albuginea at term. Routine histologic techniques and electron microscopy have been employed in characterizing the morphology of the nodules. Recent literature concerning the frequency of ovarian decidualization during pregnancy is discussed with respect to the possible relationship of such decidualization to the histogenesis of leiomyomatosis peritonealis disseminata (LPD). The hypothesis that LPD may represent "disseminated fibrosing decidua" is discussed in light of finding collagen fibrils, secretory decidual cells, and smooth muscle cells in these nodules. It is concluded that the present case does not represent "fibrosing decidua." The authors agree with others who have proposed that the smooth muscle in ovarian decidua and LPD result from proliferation of stem cells which may reside in the subperitoneal stroma in association with ectopic endometrial stroma and which may respond to the hormones of pregnancy.

Decidua

Cathepsins B1 from human fetal membranes.

Cathepsins B1 (EC 3.4.22.1) were isolated from fetal membranes of human placenta, i.e. amnion and chorion-decidua. Purification of the enzymes was achieved by the freezing-thawing technique, ammonium sulphate fractionation and Sephadex gel filtration. Cathepsis B1 separated either from amnion or from chorion-decidua exhibited optimum activity at pH 6.2, and an optimum temperature between 42-45 degrees C. They were inhibited by heavy metals, and compounds which react with the thiol groups. Isoelectric focusing demonstrated three isoenzymes of cathepsin B1 originating from chorion-decidua, while only one band was found for the enzyme from amnion.

Amnion