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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

Decidua: a possible source of amniotic fluid prolactin.

The source of amniotic fluid prolactin was investigated with the use of amnion, chorion, placenta, and decidual tissue taken from term human pregnancy. Decidua alone of these tissues contained significant quantities of prolactin. The release of decidual prolactin was affected by the presence or absence of oxygen and protein, and the amount of prolactin released far exceeded the decrease in tissue content during incubation. It is concluded that: (1) decidua may be a major source of amniotic fluid prolactin, (2) synthesis of prolactin occurs during incubation of decidua, and (3) sufficient prolactin is present in the decidua to account for that found in amniotic fluid at term.

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

Inhibitory effect of decidua on fibrinolysis induced by urokinase and by the fibrinolytic activity of the rat ovum.

With the use of a technique, which permits the study of fibrinolytic enzymes and inhibitors in organ culture, human decidua was found to produce inhibitors capable of inhibiting plasminogen activation induced by urokinase as well as activators released from kidney explants. Also the medium in which rat and human decidua had been cultured was found to inhibit fibrinolytic activity of the rat ovum. The inhibitors emanating from the decidua are presumably of importance for facilatating the implantation of ova.

Animals

The differentiation of the decidua and the distribution of metrial gland cells in the pregnant mouse uterus.

A study was made with the light microscope of the cellular changes involved in the formation of the decidua in the pregnant mouse uterus up to day 11 of pregnancy. The differentiation sequence was similar to that found by previous workers but the morphology and development of the basal zone is described in more detail. In addition, the morphology of glycogen rich cells in an area termed the lateral decidual zone is described. The distribution of granulated metrial gland cells and their precursors is described. These cells are first found in the uterine stroma before the blastocyst has implanted. Later they occur in the decidua and in the circular smooth muscle zone beneath the mesometrial triangle prior to the formation of the metrial gland. Granulated metrial gland cells are also found in the maternal blood spaces of the implantation site.

Animals

Ultrastructure of the placental villi, chorion laeve, and decidua parietalis in normal and hypertensive pregnant women.

The fine structure of the placental villi, chorion laeve, and decidua parietalis from normal and hypertensive pregnant women is described. Placental villi from hypertensive women revealed an increase in syncytial degeneration and fibrinoid deposition. Also noted were greater numbers of cytotrophoblast cells. On the other hand, there were no morphologic differences detected in the cellular and noncellular elements of the chorion laeve and decidua parietalis between normal and hypertensive subjects. Secretory granules were observed in both the placental villi and the chorion laeve.

Chorion

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

Patterns of invasive growth in vitro. Human decidua graviditatis confronted with established human cell lines and primary human explants.

Human decidua graviditatis was used as a receptor tissue to demonstrate invasive properties of human cells in vitro. In the confrontation between decidua and established cell lines of tumors and of normal origin as well as primary explanted cells, we noted differences in the cells' interactions with decidual tissue. The two original tumor cell lines, HeLa (carcinoma) and AFi (sarcoma), and a spontaneously transformed lymphoblastoid cell line showed aggressiveness, i.e., invasion, rapid proliferation, injury, and destruction of the decidual tissue. Strain HeLa S3 and two established cell lines, FL (amnion) and Girardi Heart, which are regarded as transformed cells because of their increased mitotic rate and polyploidy, exhibited various degrees of aggressiveness but did not invade. Freshly explanted fetal lung and endometrium caused no injury to the receptor tissue and were included in the decidual tissue.

Amnion

Morphological changes in the placenta and decidua after induction of abortion by extra-amniotic prostaglandin.

Seventeen placentae from second trimester abortions induced by the injection of extra-amniotic prostaglandin have been examined. The extraplacental membranes, the chorionic plate of the placenta, the placental intervillous space and the placental villi were normal in all cases. In all but one specimen lesions were, however, noted in the decidua, these including focal decidual necrosis, intradecidual haemorrhage, thrombosis of maternal vessels and dilatation and congestion of the maternal vasculature. Similar decidual lesions were also seen in placentae from spontaneous and surgical second trimester abortions and it is concluded that prostaglandins do not produce any specific lesions in the placenta or decidua. It is suggested that the negative findings of this study indicate that prostaglandins do not induce abortion by either direct injury to the placenta or by provoking utero-placental ischaemia.

Abortion, Induced

Trace elements in the human endometrium and decidua. A multielement analysis.

By means of neutron activation analysis, 25 trace elements, which are usually present in biological material, were estimated in 31 specimens of human endometrial tissue obtained at various phases of the menstrual cycle and in 14 specimens of decidua from the 12th to 18th week of pregnancy. Among the 13 trace elements invariably found in all specimens, the levels of copper, potassium, rubidium, antimony and zinc were significantly higher and those of bromine, selenium and sodium significantly lower in the endometrium than in the decidua. No difference was found in the levels of gold, calcium, cobalt, cesium and iron. Among the 12 trace elements which were found only occasionally, chromium, mercury, silver, and cadmium were detected in approximately half and cerium and scandium in approximately one-fourth of the 45 samples studied. Arsenic, barium, lanthanum, molybdenum, samarium and strontium were detected only rarely. The cyclic variations in the endometrial levels of calcium, rubidium and copper were highly significant and those in the levels of gold, cesium, iron, potassium and zinc probably significant.

Adult

The disappearance of the decidua capsularis and Reichert's membrane in the mouse.

Starting on the eleventh day the decidua capsularis and trophoblastic giant cell layer disappear from the ends of the conceptus towards the middle. The disappearance is complete antimesometrially on the eighteenth day, but lateral remnants are still present on the nineteenth day. New uterine lumen never intervenes between undegenerate decidua capsularis and the uterine stroma. Reichert's membrane is intact on the sixteenth day, but has ruptured on the seventeenth. It is suggested that its apparently earlier disappearance in histological material is an artefact reflecting the weakening that leads to its rupture.

Animals

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