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PubMed1993

An immunohistochemical study of leucocytes in human endometrium, first and third trimester basal decidua.

An immunohistochemical quantitative study of leucocyte subpopulations on fresh human endometrium and on biopsy specimens of first and third trimester basal decidua in normal (uncomplicated) pregnancies was performed. The most prominent population in endometrial and decidual stroma of basal decidua are macrophages. B cells as well as gamma/delta T cell receptor positive cells were found occasionally, scattered throughout the endometrial/decidual stroma. CD3+ cells were present in a relatively small number in the endometrium as well as in the first trimester basal decidua, but their number was elevated (doubled) in the third trimester of pregnancy. CD2+ cells showed a slight increase in first trimester basal decidua when compared with both endometrium and third trimester basal decidua. Cells with positive NKH-1 marker (CD56+) showed a significant increase in the first trimester, while in the third trimester their number diminished drastically. CD56:CD3 cell ratio increased to more than five times in first trimester basal decidua, while in the third trimester basal decidua decreased drastically. The mentioned increase of CD56+ cells in the first trimester and that of CD3+ cells at term suggests that these cells could have some specific function(s). However, it still has to be established whether the described quantitative changes of decidual leucocytes in basal decidua during pregnancy are of any importance for the mechanism(s) for the fetal allograft protection.

Antigens, CD↗
PubMed1993

Estrogen and progesterone receptors in human decidua after RU486 treatment.

OBJECTIVE: To examine RU486 action on decidua at the level of cellular estrogen receptor (ER) and P receptor (PR). DESIGN: Controlled basic study for contragestion mechanism of mifepristone. SETTING: Normal human volunteers in an academic research environment. PATIENTS: Sixty women with 6 to 7 weeks of gestation who voluntarily requested termination of pregnancy were recruited and randomly divided into three groups. INTERVENTION: A single dose of 200 mg RU486 was orally administered to the two treatment groups 12 and 24 hours, respectively, before surgical interruption of pregnancies. Placebo was used for control group. Decidual tissues were collected right after operation. MAIN OUTCOME MEASURE: Immunocytochemical reactions of PR and ER in decidua after RU486 treatment were compared with the control subjects. The differences of the reaction in decidual area with or without trophoblast invasion were noted. RESULTS: RU486 treatment increased PR and ER staining in vessel and stroma of decidua without trophoblast invasion (decidua parietalis) but not in decidua with trophoblast invasion (decidua capsularis or basalis). Chi-squared analysis indicated a significant increase in the number of ER-positive samples after RU486 treatment. CONCLUSION: The decidua parietalis was the primary target site of RU486. The lack of RU486 effect on decidua capsularis implied that trophoblast invasion prevented against antiprogestin impact.

Abortion, Induced↗
PubMed2002

Absence of HLA-G expression in macrophages of human decidua.

PROBLEM: Macrophages - together with natural killer (NK) cells - constitute the majority of bone marrow derived infiltrating cells in the decidua. As interferon-gamma (IFN-gamma), a cytokine produced by NK cells, has been reported to induce expression of human leukocyte antigen (HLA-G) in monocytic cells, suggesting expression of HLA-G on decidua macrophages potentially stimulated by IFN-gamma, the question arises whether decidua macrophages in normal pregnancy express HLA-G. METHOD OF STUDY: The study was based on immunohistochemistry and flow cytometry. In order to exclude that potentially elusive soluble HLA-G was not detected by immunohistochemistry, we performed in addition RT-PCR of flow-sorted decidua macrophages. RESULTS: Our findings indicate that HLA-G is not present on macrophages of first trimester or term decidua in either membrane-bound or soluble form. Transcripts for soluble HLA-G1 and -G2 were not detected. CONCLUSIONS: We exclude a role of HLA-G on the surface of decidua macrophages or of soluble HLA-G1 or -G2 as a secretory product of decidua macrophages with regard to interaction with HLA-G receptors present in or outside the decidua.

Cell Line↗
PubMed1999

Increased blood vessel density in decidua parietalis is associated with spontaneous human first trimester abortion.

Spontaneous pregnancy loss affects 15-18% of couples, and a number of potential causes are being discussed. The purpose of the present study was to assess if angiogenic disorders in the decidua of early human pregnancy could be related to spontaneous abortions. First trimester human decidua from elective terminations of normally progressing pregnancies and from missed abortions were investigated immunohistochemically. We quantified vessel density in decidua from normal pregnancies and from abortions by von Willebrand factor (vWF), platelet endothelial cell adhesion molecule (PECAM-1) and CD34 staining. Decidual blood vessel expression of alphavbeta3 integrin was also investigated. Significant increase (P < 0.02) in vessel density was observed in decidua parietalis of abortions, compared to decidua basalis. This increase was detected on slides stained for vWF and CD34, but not for PECAM-1. We observed a 15% increase analysing with vWF and a 77% increase with CD34 staining. alphavbeta3 integrin expression was not significantly different, neither in decidua parietalis from abortion, nor parietalis from normal pregnancies. Our data suggest that the increased vascularization in decidua parietalis from abortions could reflect complex disorders, such as specific cytokine expressions and hypoxia phenomena during the development of the decidua.

Abortion, Induced↗
PubMed1989

Characterization of an insulin-like growth factor binding protein, analogous to human pregnancy-associated secreted endometrial alpha 1-globulin, in decidua of the baboon (Papio anubis) placenta.

The major secreted protein of the human decidua (pregnancy-associated endometrial alpha 1-globulin [alpha 1-PEG]), is an insulin-like growth factor-binding protein (IGF-BP) that is immunologically and biochemically similar to placental protein 12 (PP12) extracted from term human placenta. Since previous studies have demonstrated that the baboon and human endometrium synthesize and release a number of biochemically and immunologically related polypeptides in culture, this study was undertaken to further characterize a related IGF-BP in baboon placental tissues. Decidua, chorio-amniotic membranes with adhering decidua (CAM-D), and placental villi were obtained from pregnant baboons between Days 134 and 160 of gestation by Cesarean sections. Portions of tissue were either cultured in the presence of 35S-methionine, fixed for immunocytochemistry, or frozen in liquid nitrogen for cytosol extraction. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of tissue culture media (TCM) revealed that the major secretory product of the decidua and CAM-D was an acidic polypeptide (Mr 33,000). Western blot analysis and immunoprecipitation of TCM with murine monoclonal antibody (B2H10) against human alpha 1-PEG demonstrated that this molecule, secreted by the baboon decidua and CAM-D, but not the placental villi, was immunologically identical to the human IGF-BP. Immunocytochemical localization of IGF-BP was intense in the cytoplasm of stromal cells in decidua and CAM-D and absent in the placenta. Gel filtration of TCM and cytosol followed by screening of eluates for 125I-IGF-I binding resolved two peaks (Mr greater than 100,000 and 35,000) of specific IGF-BP in decidua and CAM-D. The 35,000 peak had 100-200 times the binding capacity of the Mr greater than 100,000 peak and a Kd of 1.14-1.83 nM. The eluates contained in the Mr 35,000 peak were also immunoreactive to alpha 1-PEG, as accessed by a polyclonal radioimmunoassay. Affinity cross-linking with 125I-IGF-I followed by sodium dodecyl sulfate-PAGE revealed an immunoreactive complex of Mr 36,000, confirming that the baboon protein represents a high affinity IGF-BP. These studies indicate that the hypertrophied stromal cells of the baboon decidua and CAM-D synthesize and release an IGF-BP as their major secretory product, analogous to the situation in humans. The results of this study suggest that this protein may play a role in the regulation of IGF action during pregnancy.

Animals↗
PubMed1998

Maternal IL-11Ralpha function is required for normal decidua and fetoplacental development in mice.

In eutherian mammals, implantation and establishment of the chorioallantoic placenta are essential for embryo development and survival. As a maternal response to implantation, uterine stromal cells proliferate, differentiate, and generate the decidua, which encapsulates the conceptus and forms the maternal part of the placenta. Little is known about decidual functions and the molecular interactions that regulate its development and maintenance. Here we show that the receptor for the cytokine interleukin-11 (IL-11Ralpha) is required specifically for normal establishment of the decidua. Females homozygous for a hypomorphic IL-11Ralpha allele are fertile and their blastocysts implant and elicit the decidual response. Because of reduced cell proliferation, however, only small deciduae form. Mutant deciduae degenerate progressively, and consequently embryo-derived trophoblast cells generate a network of trophoblast giant cells but fail to form a chorioallantoic placenta, indicating that the decidua is essential for normal fetoplacentation. IL-11Ralpha is expressed in the decidua as well as in numerous other tissues and cell types, including the ovary and lymphocytes. The differentiation state and proliferative responses of B and T-lymphocytes in mutant females were normal, and wild-type females carrying IL-11Ralpha mutant ovaries had normal deciduae, suggesting that the decidualization defects do not arise secondarily as a consequence of perturbed IL-11Ralpha signaling defects in lymphoid organs or in the ovary. Therefore, IL-11Ralpha signaling at the implantation site appears to be required for decidua development.

Animals↗
PubMed1985

Selective localization of a bone marrow cell subpopulation at the implantation site in murine decidua.

Recent studies of the surface markers on cells in murine decidua have suggested that a proportion of the cells originate from the bone marrow. To examine directly the migration of bone marrow cells to decidua, we labeled whole bone marrow with TRITC and counted the number of cells appearing in decidua following adoptive transfer to virgin or allopregnant C3H mice. Labeled cells were readily identified in decidua by 3 hours after injection, and fewer cells were seen in the lining of the virgin uterus compared to allopregnant recipients. The presence of labeled cells could not be explained by blood contamination. The TRITC-labeled cells were found preferentially at the implantation sites, and while labeled bone marrow cells could be found in spleen as well as in decidua, adoptively transferred TRITC-labeled spleen cells localized preferentially in spleen and lymph nodes draining the uterus rather than in decidua. Velocity sedimentation analysis showed that the labeled bone marrow cells lodging in decidua were small lymphocytic cells (S = 2.5-4 mm/hr) and appeared to derive preferentially from small-sized bone marrow cells (S less than 4.3 mm/hr). These observations indicate that a subpopulation of bone marrow cells selectively homes to the implantation site in decidua during successful allopregnancy. The possible significance of this observation is discussed.

Animals↗
PubMed1992

Alpha 2-macroglobulin expression in the mesometrial decidua and its regulation by decidual luteotropin and prolactin.

During decidualization, cells of the endometrium grow and differentiate giving rise to two different decidual tissues located in either the antimesometrial or mesometrial site of the uterus in the rat. These tissues have different functions in pregnancy. The antimesometrial decidua is an endocrine gland that secretes hormones, whereas the mesometrial decidua appears to play an important role in limiting trophoblast invasion. Since the decidual tissue of the rat produces alpha 2-macroglobulin (alpha 2MG), we examined whether this potent protease inhibitor is specifically expressed by the mesometrial tissue, the site of trophoblast invasion, and whether the alph 2MG gene is regulated by decidual luteotropin (DLt), the PRL-like hormone secreted by the neighboring antimesometrial cells. To determine the secretory proteins of the rat decidua, antimesometrial and mesometrial tissues were dissected out from pseudopregnant rats and cultured with [35S]methionine. The major protein secreted by the antimesometrial cell was the 29-kilodalton decidual luteotropin, whereas a 180-kilodalton protein was predominantly secreted by the mesometrial tissue. Immunoprecipitation studies of 35S-radiolabeled proteins revealed that this high mol wt protein is alpha 2MG and that it is secreted exclusively by the cells forming the mesometrial tissue. To examine whether the alpha 2MG gene was also expressed specifically in the mesometrial decidua, total RNA was isolated from both mesometrial and antimesometrial tissues of day 9-12 pseudopregnant rats and hybridized with alpha 2MG cDNA. Northern blot analysis revealed a 5.4-kilobase message, which was abundantly expressed in the mesometrial decidua. Little, if any, alpha 2MG mRNA was detected in antimesometrial decidua. The ontogeny of the message in the decidua correlated well with the development of the mesometrial tissue. To examine whether alpha 2MG expression is regulated by DLt and/or PRL, a highly specific polyclonal antibody to DLt was generated and decidual tissues were cultured in the presence or absence of DLt antibodies with or without PRL. Neutralization of DLt caused a marked decrease in alpha 2MG mRNA levels. This down-regulation was totally reversed by the addition of PRL and was not affected by alpha 2MG antibodies. In summary, the results of this investigation revealed a compartmentalized gene expression, synthesis and secretion of alpha 2MG in the decidua. The secretion of this protease inhibitor, specifically by the mesometrial tissue which is the site of trophoblast invasion, may be the reason for the minimal amount of tissue damage that occurs during placentation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1984

Formation of unconjugated estrogens from estrone sulfate by dispersed cells from human fetal membranes and decidua.

Experiments were performed to determine the ability of isolated cells from human amnion, chorion, and decidua to hydrolyze estrone sulfate (E1S) to free estrone (E1) and estradiol (E2). Tissues were obtained from eight women after spontaneous onset of labor and vaginal delivery and from eight women undergoing elective cesarean section at term before the onset of labor. The tissues were obtained immediately after delivery, dispersed into isolated cell preparations using 0.05% collagenase, and incubated in Krebs-Ringer bicarbonate at 37 degrees C for 4 h. E1 and E1S were measured by specific RIAs. All three tissues from both patient groups produced E1 and E2 in a dose-dependent fashion from the E1S precursor. In both patient groups, chorion and decidua cells produced significantly more E1 and E2 than amnion cells. The chorion cells from the spontaneous labor group produced significantly more E1 than chorion cells from the cesarean section group. The chorion and decidua cells from the spontaneous labor group produced significantly more E2 than corresponding cells from the cesarean section group. The hydrolysis of E1S to E1 and E2 was significantly inhibited in a dose-dependent fashion by increasing concentrations of oxytocin and (Bu)2cAMP. Dispersed cells from amnion, chorion, and decidua also converted [3H]E1S to radiochemically pure [3H]E1. Chorion and decidua cells were significantly more active in this respect than amnion. The presence of excessive amounts of dehydroepiandrosterone sulfate (DHAS) did not significantly alter this reaction. All three tissues also hydrolyzed [3H]dehydroepiandrosterone sulfate, but in chorion and decidua, this activity was significantly less than the hydrolysis of [3H]E1S. We conclude that human amnion, chorion, and decidua possess the capability to hydrolyze E1S to free estrogen and that this activity in chorion and decidua is increased after spontaneous vaginal delivery. It is possible that this activity and its regulation are important factors in the local synthesis of free estrogen, which, in turn, may influence myometrial contractility.

Adult↗
PubMed1987

The inhibitory effect of cytosolic fraction of human decidua on prostaglandin synthesis.

To elucidate the mechanism of suppression of prostaglandin (PG) production in decidua in early pregnancy, the PG synthetase activity of decidua and mid-secretory endometrium was studied. The microsomal fractions and their supernatants were prepared from the tissue by ultracentrifugation at 105,000 g. The standard incubation mixture consisted of the microsomal fraction and 14C arachidonic acid with cofactors, with incubation being carried out for 10 minutes at 37 degrees C. After extraction, the radioactivity of PGE2 was measured and PG synthetase activity was assayed. The apparent Km value for PG synthetase in decidua was 4.6 +/- 0.14 x 10(-6) M (n = 4), whereas that in endometrium was 4.6 +/- 1.18 x 10(-6) M (n = 3). Subsequently, kinetic studies on PG synthetase inhibitor in decidua were carried out. When sheep seminal vesicle was used as an enzyme source, the decidual supernatant showed competitive inhibition. The inhibitory substance in decidua was inactivated after incubation for 15 minutes at 65 degrees C. It seems likely that the suppression of PG biosynthesis in human decidua in early pregnancy is not due to the difference in PG synthetase found in decidua and in endometrium, but due to the existence of PG synthetase inhibitor in decidua.

Cell Extracts↗
PubMed1989

Immunohistological localization of human pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG), a glycosylated beta-lactoglobulin homologue, in the decidua and placenta during pregnancy.

Monoclonal and polyclonal antibodies to pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG), a glycosylated human beta-lactoglobulin homologue, were used in an immunohistological technique to determine the cellular localization of this protein in the decidua and placental tissues during pregnancy. During the first trimester the protein was principally localized to the glandular epithelium of the decidua spongiosa region of the endometrium with only weak staining associated with glands of the decidualized decidua compacta region. No significant cellular staining was detected in the decidua capsularis. At term in the decidua of the amniochorion and the placental bed weak staining for alpha 2-PEG was only associated with the epithelium of attenuated glands. No significant staining was detected in the placenta during pregnancy. These results suggest that the epithelium of glands associated with non-decidualized stroma represents the primary source of alpha 2-PEG during the first trimester and that a function of the decidua spongiosa in early pregnancy may be related to production of alpha 2-PEG. The decline in production of alpha 2-PEG during pregnancy is suggested to result from involution of the decidua spongiosa and at term the attenuated glands of the decidua represents the source of alpha 2-PEG.

Decidua↗
PubMed1990

Prostaglandin-mediated inactivation of natural killer cells in the murine decidua.

Previous studies from this laboratory have demonstrated a large influx of null lymphocytes into the murine decidua during pregnancy. We had also shown that trophoblast cells of the murine placenta bear target structures recognized by NK cells. Since NK lineage cells belong to the null category of lymphocytes, we examined whether cells of this lineage appear in the murine decidua, and if so, whether their activity is locally regulated by NK suppressor cells. We further investigated the identity of the suppressor cells as well as their suppressor products. NK lineage cells, irrespective of their activation status, were identified morphologically in radioautographic preparations as the non-T, non-B (null) lymphocytes capable of binding YAC-1 lymphoma targets. NK activity of nucleated cells was measured with a 4-hr 51Cr-release assay against labeled YAC-1 targets. Studies with outbred CD1 mice, and to a smaller extent, inbred CBA mice revealed that the incidence of NK lineage cells remained fairly constant within the decidua throughout pregnancy, but their activity decreased steadily to negligible levels by Day 12-14 of gestation. This was found to result from an inactivation caused by NK-suppressor cells in the decidua. A mixing of Ficoll-Paque-separated nucleated cells of the decidua with normal splenic effector cells (at 1:1 ratio) led to a suppression of their NK activity tested immediately or after a 20-hr coculture. This suppression was MHC unrestricted. Suppressor cells were identified both in plastic nonadherent fraction highly enriched for typical decidual cells as well as in the plastic adherent fraction containing decidual cells and macrophages. Addition of indomethacin (10(-5) M), an inhibitor of prostaglandin synthesis, or anti PGE2 antibody, revived the NK activity in the mixed population, as well as in the decidua, suggesting a PGE2-mediated suppression. High levels of PGE2 were detectable in decidual cell supernatants with a sensitive radioimmunoassay. Addition of pure PGE2 (10(-7)-10(-6) M) but not PGF2 alpha (10(-6) M) during the NK assay or to the effector cells for a 20-hr period prior to the assay led to an inhibition of NK activity. These results reveal that NK cells appearing in the murine decidua are progressively inactivated by PGE2 produced by decidual cells and decidual macrophages.

Animals↗
PubMed2001

Leukocyte activation in the decidua of chromosomally normal and abnormal fetuses from women with recurrent abortion.

As part of our continuing programme to investigate immunological causes of unexplained recurrent pregnancy losses, we studied subpopulations of white blood cells and their activation status in decidua of women with a history of recurrent spontaneous abortion (RSA). We differentiated specifically between normal karyotyped male fetuses and abnormal karyotyped fetuses with trisomy 16 because trisomy 16 is not compatible with life and is thus a non-controversial cause of spontaneous miscarriage. Leukocytes were counted in paraffin-embedded decidua after immunohistological staining for CD45 (LCA), CD3, CD56, CD68, CD69 and CD25. Numbers of activated versus non-activated T lymphocytes, NK cells and macrophages were compared in decidua from women with: (i) unexplained RSA who had a normal male karyotype (n = 17) miscarriage; (ii) unexplained RSA who had a trisomy 16 (n = 21) miscarriage; and (iii) normal gestationally age-matched first trimester pregnancies following elective termination procedures (n = 20). Significantly more activated leukocytes were detected in the decidua of women with unexplained RSA who had a normal male karyotype compared to the other groups (P < 0.0001). In addition, numbers of cells comprising the major leukocyte subpopulation, CD56+ NK cells, appeared reduced in the decidua of women with unexplained RSA compared to decidua from women having elective terminations. Increased numbers of activated leukocytes in the decidua of women with a history of unexplained recurrent pregnancy loss who had a normal karyotyped pregnancy provide evidence that cellular immunity may be involved in unexplained recurrent pregnancy loss.

Abortion, Habitual↗
PubMed2001

Serglycin proteoglycan synthesis in the murine uterine decidua and early embryo.

This study has explored the localization and synthesis of the serglycin proteoglycan in the murine embryo and uterine decidua during midgestation. Embryos in deciduae were subjected to in situ hybridization with cRNA probes and to immunohistochemical detection with a specific antibody against murine serglycin. Adherent decidual cell cultures were prepared from freshly isolated deciduae. Proteoglycan biosynthesis was investigated by labeling intact deciduae and decidual cultures with (35)S-sulfate. Serglycin mRNA was detected by in situ hybridization throughout the mesometrial portion and at the periphery of the antimesometrial portion of the decidua at Embryonic Day (E) 8.5, and in the parietal endoderm surrounding the embryo. Serglycin mRNA was detected in fetal liver at E11.5-E14.5. Serglycin was detected by immunohistochemistry in decidua and parietal endoderm at E8.5 and in liver at E13.5. Most of the proteoglycans synthesized by cultured intact deciduae (78%) and adherent decidual cultures (91%) were secreted into the medium. Serglycin proteoglycan may play an important role in uterine decidual function during early postimplantation development.

Animals↗
PubMed1996

Comparison of immunosuppressive properties of hydatidiform mole decidua and trophoblast extracts.

PROBLEMS: The immunologic privilege afforded the fetus relies upon immunoregulation within the maternal-fetal interface. Trophoblast and decidua-derived immunoregulatory factors enforce this privilege by locally suppressing maternal responses to trophoblast antigens. The relative contribution of trophoblast or decidua immunosuppressive factors to pregnancy immunotolerance are not well characterized. The purpose of this study was to compare the suppressive effects of hydatidiform mole trophoblast and decidua extracts on interleukin-2-dependent proliferation. METHOD: Tissue extracts were prepared from hydatidiform mole trophoblast and decidua following uterine evacuation. Samples were submitted to interleukin-2-dependent and -independent cell proliferation assays. RESULTS: Hydatidiform mole trophoblast extract significantly (P < 0.05) suppressed interleukin-2-dependent proliferation but did not affect interleukin-2-dependent cell proliferation. In contrast, molar decidua extract suppressed both cell lines. CONCLUSIONS: Human hydatidiform mole trophoblast contains factor(s) that specifically abrogate interleukin-2-dependent clonal expansion of murine cytotoxic T-cells. In contrast, extracts of molar decidua suppressed both interleukin-2-dependent and -independent responses. This indicates that the trophoblast antagonizes critical interleukin-2-mediated immunologic responses, but that the decidua uses nonspecific antiproliferative mechanisms for immunoregulation.

Decidua↗
PubMed2000

Human first-trimester decidua vascular density: an immunohistochemical study using VE-cadherin and endoglin as endothelial cell markers.

PROBLEM: Angiogenesis and vasculogenesis appear to be of critical importance for the success of pregnancy. Recent data have emphasized that pregnancy complications, such as abortion or pre-eclampsia, are linked with vascular pathologies. The aim of this study was to quantify human first-trimester decidua microvascular density, using two novel, highly specific endothelial cell markers, VE-cadherin and endoglin. METHOD OF STUDY: We collected decidua from women undergoing termination of normal pregnancies. VE-cadherin and endoglin were localized by immunohistochemistry. The blood vessel densities detected by VE-cadherin or endoglin-stainings were microscopically quantified per mm2. RESULTS: Endothelial cells in first-trimester human decidua both express VE-cadherin and endoglin. The microvascular density detected by VE-cadherin-staining varied from 32.2 +/- 1.7 in decidua basalis, to 30 +/- 0.6 in decidua parietalis. For the endoglin-staining, the values varied from 37.5 +/- 3 in decidua basalis, and 26.7 +/- 1.2 in decidua parietalis. CONCLUSIONS: Our data shows that both VE-cadherin and endoglin are good candidates to highlight the decidual endothelial cells, and to quantify the blood vessels density of endometrium.

Abortion, Induced↗
PubMed1996

The decidual prolactin receptor and its regulation by decidua-derived factors.

Decidualization of the endometrial stroma in the rat gives rise to two different cell populations, located either mesometrially or antimesometrially in the uterus. We have previously shown that the rat decidua is the site of production and action of a PRL-like hormone. In this investigation we examined, using reverse transcription-PCR, whether and which type of PRL receptor (PRL-R) messenger RNA (mRNA) is expressed in the decidua, whether the receptor is confined only to one cell population, and whether the PRL-R expression is regulated by decidua-derived factors. The results indicate that the uterus of pseudopregnant rats does not express the PRL-R and that decidualization does not trigger a rapid appearance of PRL-R mRNA. It is only 3 days after the induction of decidualization that the long form of the PRL-R was first expressed. Thereafter, mRNAs for both the short (PRL-RS) and the long (PRL-RL) form became detectable in both antimesometrial and mesometrial decidua, although PRL-RL mRNA was much more abundant than PRL-RS. As development proceeded, PRL-R mRNA decreased and disappeared specifically from the antimesometrial decidua, whereas the mesometrial decidua continued to express this receptor mRNA. Concomitant with down-regulation of the PRL-R in the antimesometrial tissue was a rather abrupt expression of activin A. In contrast, the mesometrial tissue that maintained high levels of PRL-R mRNA expressed little activin A, but produced an activin-binding protein, alpha(2)-macroglobulin (alpha(2)MG). To determine whether activin A and alpha(2)MG regulate PRL-R expression, antimesometrial and mesometrial cells were separated by elutriation and maintained in culture in the presence or absence of activin A, alpha(2)MG, or follistatin. Just after cell separation, both cell populations expressed PRL-R, but not activin A. Within 6 h, activin A mRNA and protein became highly expressed in the mesometrial cells, whereas PRL-RL mRNA became undetectable. In contrast, activin A mRNA was at very low levels in the antimesometrial cells, and no activin A protein could be detected in the medium for at least 12 h. In these cells PRL-RL mRNA remained elevated. Addition of activin A to antimesometrial cells caused a marked down-regulation of PRL-RL mRNA expression, whereas addition of alpha(2)MG and follistatin to mesometrial cells prevented the disappearance of PRL-R. In summary, the results of this investigation 1) indicate that decidualization of the endometrial stroma induces the appearance of both forms of the PRL-R mRNAs; 2) show differential expression of the PRL-R mRNA in the two-cell population forming the decidua; 3) establish that this differential expression is due to two key decidual molecules, activin A and alpha(2) macroglobulin; and 4) demonstrate that activin A can cause the decidual cells to lose the PRL-R and that the disappearance of the decidual PRL-R can be prevented by addition to the culture of two activin binding proteins, follistatin and alpha(2) MG.

Activins↗
PubMedDate not supplied

Progesterone receptor localization and isoforms in myometrium, decidua, and fetal membranes from rhesus macaques: evidence for functional progesterone withdrawal at parturition.

OBJECTIVE: It is not known whether withdrawal of progesterone (P) action is a prerequisite for parturition in women or in nonhuman primates because concentrations of circulating progesterone or progesterone receptors (PR) in myometrium and decidua do not decrease before delivery. To examine this potentially important regulatory mechanism, we determined PR isoforms, PR localization, and mRNA in myometrium, decidua, and fetal membranes from rhesus monkeys during pregnancy and in spontaneous labor at term. METHODS: Gestational tissues were obtained midpregnancy (day 80-100), late pregnancy (day 130-145), and during spontaneous labor at term (day 161-167). Samples of rhesus monkey myometrium, decidua, chorion-decidua, and amnion were collected and analyzed for total nuclear and cytosolic PR by competitive binding assay. Progesterone receptor isoforms were identified and quantified by Western blot analysis, and PR mRNA was determined by a specific ribonuclease protection assay. Nuclear PR was localized by immunohistochemistry with monoclonal anti-PR (JZB39) after microwave stabilization. RESULTS: Myometrium and decidua showed no change in total PR during pregnancy and labor. Nuclear PR was not detected in fetal membranes by binding assay but was localized in amnion epithelial and mesenchymal cells and in chorion laeve cytotrophoblasts by immunohistochemistry. Staining for PR was substantially less by serial antibody dilution in fetal membranes than in decidua. Message for PR was confirmed in all tissues analyzed. A significant (P <.05) shift in the ratio of PR isoforms (from PR-B dominance at midpregnancy to PR-A dominance in labor) was observed in myometrium but not in decidua. Both PR-A and PR-B isoforms and PR nuclear staining were nearly undetectable in amnion obtained during labor. CONCLUSION: A shift to PR-A dominance in myometrium at term together with a loss of PR in fetal membranes provides evidence for a functional progesterone withdrawal mechanism, which may facilitate the initiation of parturition in primates.

Animals↗