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Investigation of calcium transport and regulation system in human placenta
Abstracts / Placenta 36 (2015) A1eA14
the mouse placenta. However, there is little information available on the expression pattern of this ncRNA in placental development. We examined the expression level of H19 ncRNA in the mouse placenta by real-time quantitative reverse transcription PCR (real-time PCR) and in situ hybridization. Methods: We extracted total RNA from B6D2F1 mouse placentas at five different developmental stages (i.e., E7.5, E10.5, E13.5, E16.5, and E18.5) and adult mouse organs (i.e., heart, lung, liver, kidney, intestine, spleen, uterus, and ovary). The expression level of H19 ncRNA was investigated by real-time PCR. Moreover, in situ hybridization was performed to identify the cell types expressing H19 ncRNA in the placenta using digoxigeninlabelled RNA probes. Results: Real-time PCR revealed that H19 ncRNA was highly expressed in the placenta compared to other organs. Although H19 was detected at E7.5, it was significantly upregulated at the later stages of placental development. In situ hybridization showed that H19 was present in glycogen cells in the decidua basalis, trophoblast giant cells in the junctional zone, and mononuclear trophoblasts in the labyrinth zone of the mouse placenta. Conclusion: H19 ncRNA was markedly expressed in the late stages of mouse placental development. Our findings may provide insights into the role of H19 ncRNA in the mouse placenta.
JPA2015-24. MOUSE PLACENTAL DISTRIBUTION OF ENZYMES AND TRANSPORTER MODULATING PGE2 LEVEL Mai Inagaki 1, Tomohiro Nishimura 1, Takeo Nakanishi 2, Shin-ichi Masanori Tachikawa 4, Ikumi Tamai 2, Ken-ichi Akanuma 3, Hosoya 3, Masatoshi Tomi 1, Emi Nakashima 1. 1 Faculty of Pharmacy, Keio University, Japan; 2 Faculty of Pharmaceutical Sciences, Kanazawa University, Japan; 3 Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Japan; 4 Graduate School of Pharmaceutical Sciences, Tohoku University, Japan Objectives: Placenta-derived prostaglandin (PG) E2 plays important roles in patent ductus arteriosus in the fetus, trophoblast invasion, and induction of labor. It is thought that PGE2 level in the placenta is strictly regulated by enzymes and transporters such as cyclooxygenase (COX) and microsomal PGE synthase-1 (mPGES-1), 15-hydroxyPG dehydrogenase (15-PGDH), and PG transporter (PGT/SLCO2A1). Information about localizations of these proteins leads to a further understanding of the mechanism for regulating PGE2 signaling in the placenta. The purpose of this study is to clarify the localizations of proteins regulating PGE2 distribution in the mouse placenta. Methods: The placenta was dissected into three areas: the decidua, the junctional zone, and the labyrinth zone. Western blot analysis was performed to measure the expression levels of COX isoforms and mPGES-1 from mid-gestation to term. Immunofluorescence analysis was performed to demonstrate localizations of COX isoforms, mPGES-1, 15-PGDH, and PGT. PGE2 concentration was determined by EIA assay. Results: The expression levels of COX-1 and mPGES-1 were gradually increased toward term in the labyrinth zone. The signal for mPGES-1 in the labyrinth zone was detected in the fetal facing syncytiotrophoblasts, and was overlapped with the signal for COX-1. On the other hand, the signal for COX-2 was not overlapped with that for mPGES-1. PGT and 15-PGDH were co-localized in spongiotrophoblasts in the junctional zone. Of three placental areas, PGE2 concentration was the lowest in the junctional zone and was the highest in the labyrinth zone. Conclusion: These data suggest that the fetal-facing syncytiotrophoblasts release PGE2 into the fetal circulation via COX-1 and mPGES-1-mediated PGE2 production, and that spongiotrophoblasts regulate PGE2 level in the placenta via PGT-mediated PGE2 uptake and 15-PGDH-mediated oxidation of PGE2.
JPA2015-26. INVESTIGATION OF CALCIUM TRANSPORT AND REGULATION SYSTEM IN HUMAN PLACENTA Toshihide Sakuragi, Yoko Aoyama, Yukiyo Aiko, Chiharu Tomonaga, Satoshi Aramaki, Hirohide Inagaki, Kimi Nakashima, Toru
A7
Hachisuga, Eiji Shibata. Department of Obstetrics and Gynecology, University of Occupational and Environmental Health, Japan The TRPV (Transient Receptor Potential Vanilloid) 5 and 6 are members of a highly conserved family of Calcium channels sensitive to intracellular Ca2+. TRPV 5 and 6 are both highly selective for Ca2+ as well as certain metal ions including Cadmium and Zinc. TRPV 5 and 6 are expressed in the placenta, and are thought to be the primary Ca2+ channels of the syncytiotroplast, and responsible for active Ca2+ transport across the placenta in cooperation with Calbindin and PMCA ATPase. Ca2+ transport is critical for proper fetal development, particularly bone calcification. Recent reports describe reduced activity and expression of these Ca2+ channels in the trophoblast of Preeclampsia-associated placenta. However, only TRPV6 has been shown to be expressed in the syncytiotrophoblast. Using immunohistochemistry of placenta and kidney, we found that TRPV 5 protein expression is primarily expressed in the cytotrophoblasts of the microvillous. TRPV5 staining in the syncytiotrophblast was very limited, primarily restricted to the nuclear syncytium. Therefore, the localization of TRPV5 with the placental microvillous is quite different from TRPV 6. Calbindin was expressed in the cytoplasm of syncytiotrophblast. PMCA ATPase was localized in the basal membrane of syncytiotrophblast. Those expression patterns were similar to those of distal convoluted renal tubule. In addition, our preliminary studies found no significant differences in TRPV 5 and 6 expression levels in the placenta of Preeclampsia or IUGRassociated pregnancies compared to controls. Our data suggests that TRPV5 and TRPV6 have distinct roles in the placenta like a kidney.
JPA2015-27. A CASE OF A PREGNANT WOMAN WITH A MASSIVE SUBCHORIONIC HEMATOMA AND FETAL GROWTH RESTRICTION DETECTED IN THE SECOND TRIMESTER Takanori Yokoyama, Norifumi Tanaka, Hiroshi Miyoshi, Yoshiki Kudo. Department of Obstetrics and Gynecology, Graduate School of Biomedical Sciences, Hiroshima University, Japan Massive subchorionic hematoma is associated with poor pregnancy outcome such as fetal growth restriction (FGR), non reassuring fetal status (NRFS), intrauterine fetal death (IUFD), and premature labor. So it is very important to manage carefully and to determine the timing of the termination. We describe a woman with a diagnosis of massive subchorionic hematoma and FGR. She was a 31-year-old nulliparous woman. Because of irregular genital bleeding, she had been hospitalization in her previous hospital for a week at 26-27 weeks of gestation. An abnormal ultrasonographic finding like a subchorionic hematoma was detected during the hospitalization. When she was referred to our hospital at 27 weeks of gestation, we detected placentomegaly and FGR, and make her admit to our hospital. Then subsequent ultrasonographic examinations revealed the placentomegaly as a massive subchorionic hematoma beneath the attachment site of umbilical cord. At 28 weeks of gestation, because we found CTG-sign sporadic late decelerations, cesarean section was performed. The infant weighed 974 g. The apgar score was 9 at 1 min and 10 at 5 min. Fresh infarction and fibrin deposition were found under chorionic plate histopathologically.
JPA2015-28. A CASE OF PLACENTAL ENLARGEMENT IN THE SECOND TRIMESTER Megumi Ueda, Chizuko Yaguchi, Naomi Huruta, Kazuhiro Sugihara, Hiroaki Itoh, Naohiro Kanayama. Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Japan We reported a case of a stillborn in 24 gestational weeks with abnormal placental enlargement confirmed by ultrasonography. 33-year-old primiparous woman, who was impregnated through in vitro fertilization, was referred to our hospital at 11 weeks for abnormal vaginal bleeding and a wide range of subchorionic hematoma. After natural improvement of subchorionic hematoma at 19 weeks, she was
the mouse placenta. However, there is little information available on the expression pattern of this ncRNA in placental development. We examined the expression level of H19 ncRNA in the mouse placenta by real-time quantitative reverse transcription PCR (real-time PCR) and in situ hybridization. Methods: We extracted total RNA from B6D2F1 mouse placentas at five different developmental stages (i.e., E7.5, E10.5, E13.5, E16.5, and E18.5) and adult mouse organs (i.e., heart, lung, liver, kidney, intestine, spleen, uterus, and ovary). The expression level of H19 ncRNA was investigated by real-time PCR. Moreover, in situ hybridization was performed to identify the cell types expressing H19 ncRNA in the placenta using digoxigeninlabelled RNA probes. Results: Real-time PCR revealed that H19 ncRNA was highly expressed in the placenta compared to other organs. Although H19 was detected at E7.5, it was significantly upregulated at the later stages of placental development. In situ hybridization showed that H19 was present in glycogen cells in the decidua basalis, trophoblast giant cells in the junctional zone, and mononuclear trophoblasts in the labyrinth zone of the mouse placenta. Conclusion: H19 ncRNA was markedly expressed in the late stages of mouse placental development. Our findings may provide insights into the role of H19 ncRNA in the mouse placenta.
JPA2015-24. MOUSE PLACENTAL DISTRIBUTION OF ENZYMES AND TRANSPORTER MODULATING PGE2 LEVEL Mai Inagaki 1, Tomohiro Nishimura 1, Takeo Nakanishi 2, Shin-ichi Masanori Tachikawa 4, Ikumi Tamai 2, Ken-ichi Akanuma 3, Hosoya 3, Masatoshi Tomi 1, Emi Nakashima 1. 1 Faculty of Pharmacy, Keio University, Japan; 2 Faculty of Pharmaceutical Sciences, Kanazawa University, Japan; 3 Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Japan; 4 Graduate School of Pharmaceutical Sciences, Tohoku University, Japan Objectives: Placenta-derived prostaglandin (PG) E2 plays important roles in patent ductus arteriosus in the fetus, trophoblast invasion, and induction of labor. It is thought that PGE2 level in the placenta is strictly regulated by enzymes and transporters such as cyclooxygenase (COX) and microsomal PGE synthase-1 (mPGES-1), 15-hydroxyPG dehydrogenase (15-PGDH), and PG transporter (PGT/SLCO2A1). Information about localizations of these proteins leads to a further understanding of the mechanism for regulating PGE2 signaling in the placenta. The purpose of this study is to clarify the localizations of proteins regulating PGE2 distribution in the mouse placenta. Methods: The placenta was dissected into three areas: the decidua, the junctional zone, and the labyrinth zone. Western blot analysis was performed to measure the expression levels of COX isoforms and mPGES-1 from mid-gestation to term. Immunofluorescence analysis was performed to demonstrate localizations of COX isoforms, mPGES-1, 15-PGDH, and PGT. PGE2 concentration was determined by EIA assay. Results: The expression levels of COX-1 and mPGES-1 were gradually increased toward term in the labyrinth zone. The signal for mPGES-1 in the labyrinth zone was detected in the fetal facing syncytiotrophoblasts, and was overlapped with the signal for COX-1. On the other hand, the signal for COX-2 was not overlapped with that for mPGES-1. PGT and 15-PGDH were co-localized in spongiotrophoblasts in the junctional zone. Of three placental areas, PGE2 concentration was the lowest in the junctional zone and was the highest in the labyrinth zone. Conclusion: These data suggest that the fetal-facing syncytiotrophoblasts release PGE2 into the fetal circulation via COX-1 and mPGES-1-mediated PGE2 production, and that spongiotrophoblasts regulate PGE2 level in the placenta via PGT-mediated PGE2 uptake and 15-PGDH-mediated oxidation of PGE2.
JPA2015-26. INVESTIGATION OF CALCIUM TRANSPORT AND REGULATION SYSTEM IN HUMAN PLACENTA Toshihide Sakuragi, Yoko Aoyama, Yukiyo Aiko, Chiharu Tomonaga, Satoshi Aramaki, Hirohide Inagaki, Kimi Nakashima, Toru
A7
Hachisuga, Eiji Shibata. Department of Obstetrics and Gynecology, University of Occupational and Environmental Health, Japan The TRPV (Transient Receptor Potential Vanilloid) 5 and 6 are members of a highly conserved family of Calcium channels sensitive to intracellular Ca2+. TRPV 5 and 6 are both highly selective for Ca2+ as well as certain metal ions including Cadmium and Zinc. TRPV 5 and 6 are expressed in the placenta, and are thought to be the primary Ca2+ channels of the syncytiotroplast, and responsible for active Ca2+ transport across the placenta in cooperation with Calbindin and PMCA ATPase. Ca2+ transport is critical for proper fetal development, particularly bone calcification. Recent reports describe reduced activity and expression of these Ca2+ channels in the trophoblast of Preeclampsia-associated placenta. However, only TRPV6 has been shown to be expressed in the syncytiotrophoblast. Using immunohistochemistry of placenta and kidney, we found that TRPV 5 protein expression is primarily expressed in the cytotrophoblasts of the microvillous. TRPV5 staining in the syncytiotrophblast was very limited, primarily restricted to the nuclear syncytium. Therefore, the localization of TRPV5 with the placental microvillous is quite different from TRPV 6. Calbindin was expressed in the cytoplasm of syncytiotrophblast. PMCA ATPase was localized in the basal membrane of syncytiotrophblast. Those expression patterns were similar to those of distal convoluted renal tubule. In addition, our preliminary studies found no significant differences in TRPV 5 and 6 expression levels in the placenta of Preeclampsia or IUGRassociated pregnancies compared to controls. Our data suggests that TRPV5 and TRPV6 have distinct roles in the placenta like a kidney.
JPA2015-27. A CASE OF A PREGNANT WOMAN WITH A MASSIVE SUBCHORIONIC HEMATOMA AND FETAL GROWTH RESTRICTION DETECTED IN THE SECOND TRIMESTER Takanori Yokoyama, Norifumi Tanaka, Hiroshi Miyoshi, Yoshiki Kudo. Department of Obstetrics and Gynecology, Graduate School of Biomedical Sciences, Hiroshima University, Japan Massive subchorionic hematoma is associated with poor pregnancy outcome such as fetal growth restriction (FGR), non reassuring fetal status (NRFS), intrauterine fetal death (IUFD), and premature labor. So it is very important to manage carefully and to determine the timing of the termination. We describe a woman with a diagnosis of massive subchorionic hematoma and FGR. She was a 31-year-old nulliparous woman. Because of irregular genital bleeding, she had been hospitalization in her previous hospital for a week at 26-27 weeks of gestation. An abnormal ultrasonographic finding like a subchorionic hematoma was detected during the hospitalization. When she was referred to our hospital at 27 weeks of gestation, we detected placentomegaly and FGR, and make her admit to our hospital. Then subsequent ultrasonographic examinations revealed the placentomegaly as a massive subchorionic hematoma beneath the attachment site of umbilical cord. At 28 weeks of gestation, because we found CTG-sign sporadic late decelerations, cesarean section was performed. The infant weighed 974 g. The apgar score was 9 at 1 min and 10 at 5 min. Fresh infarction and fibrin deposition were found under chorionic plate histopathologically.
JPA2015-28. A CASE OF PLACENTAL ENLARGEMENT IN THE SECOND TRIMESTER Megumi Ueda, Chizuko Yaguchi, Naomi Huruta, Kazuhiro Sugihara, Hiroaki Itoh, Naohiro Kanayama. Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Japan We reported a case of a stillborn in 24 gestational weeks with abnormal placental enlargement confirmed by ultrasonography. 33-year-old primiparous woman, who was impregnated through in vitro fertilization, was referred to our hospital at 11 weeks for abnormal vaginal bleeding and a wide range of subchorionic hematoma. After natural improvement of subchorionic hematoma at 19 weeks, she was