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      • Identification of differentially expressed genes in the uterine endometrium on day 12 of the estrous cycle and pregnancy in pigs

        Ka, Hakhyun,Seo, Heewon,Kim, Mingoo,Choi, Yohan,Lee, Chang-Kyu Wiley Subscription Services, Inc., A Wiley Company 2009 Molecular reproduction and development Vol.76 No.1

        <P>Maternal recognition of pregnancy in pigs occurs approximately on Day (D) 12 of pregnancy and is critical for embryo implantation. The presence of the conceptus in the uterine lumen during this period changes uterine endometrial function to prepare for attachment of the conceptus to the endometrial epithelial cells and maintain luteal function in the ovary. Although much is known about endometrial gene expression, the genes expressed in the uterine endometria and the cellular and molecular mechanisms of those gene products during the period of implantation and maternal recognition of pregnancy in pigs are still not completely defined. To better understand the interactions between the maternal uterus and conceptus during the implantation process, we searched genes differentially expressed in the endometria on D12 of pregnancy compared to those on D12 of the estrous cycle. A new reverse transcription-polymerase chain reaction (RT-PCR)-based method that involves annealing control primers (ACPs) was employed. Using 120 ACPs, we sequenced 12 differentially expressed genes (DEGs) and identified those genes using the Basic Local Alignment Search Tool (BLAST). Northern blot hybridization analysis confirmed the differential expression of those DEGs in the uterine endometrium. In situ hybridization analysis determined the cell-type specific expression of the DEGs in the uterine endometrium. Further analysis of the DEGs found in this study will provide insights into the cellular and molecular basis of maternal and fetal interactions during the period of maternal recognition of pregnancy in the pig. Mol. Reprod. Dev. 76: 75–84, 2009. © 2008 Wiley-Liss, Inc.</P>

      • Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand Activates Pro-Survival Signaling Pathways, Nuclear Factor-${\kappa}B$ and Extracellular Signal-Regulated Kinase 1/2 in Trophoblast Cell Line, JEG-3

        Ka Hakhyun The Korean Society of Animal Reproduction 2005 Reproductive & developmental biology Vol.29 No.2

        Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) is a well-known inducer of apoptotic cell death in many tumor cells. 1RAIL is expressed in human placenta, and cytotrophoblast cells express 1RAIL receptors. However, the role of TRAIL in human placentas and cytotrophoblast cells is not. well understood. In this study a trophoblast cell line, JEG-3, was used as a model system to examine the effect of TRAIL. on key intracellular signaling pathways involved in the control of trophoblastic cell apoptosis and survival JEG-3 cells expressed receptors for 1RAIL, death receptor (DR) 4, DR5, decoy receptor (OcR) 1 and DeR2. Recombinant human TRAIL (rhTRAIL) did not have a cytotoxic effect determined by MIT assay and did not induce apoptotic cell death determined by poly-(ADP-ribose) polymerase cleavage assay. rhTRAIL induced a rapid and transient nuclear translocation of nuclear $factor-{\kappa}B(NF-{\kappa}B)$ determined by immunoblotting using nuclear protein extracts. rhTRAIL rapidly activated extracellular signal-regulated protein kinase (ERK) 1/2 as determined by immnoblotting for phospho-ERK1/2. However, c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38MAPK) and Akt (protein kinase B) were not activated by rhTRAIL. The ability of 1RAIL to induce $NF-{\kappa}B$ and ERK1/2 suggests that interaction between TRAIL and its receptors may play an important role in trophoblast cell function during pregnancy.

      • KCI등재후보

        Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand Activates Pro-Survival Signaling Pathways, Nuclear Factor-B and Extracellular Signal-Regulated Kinase 1/2 in Trophoblast Cell Line, JEG-3

        Hakhyun Ka 사단법인 한국동물생명공학회 2005 Reproductive & developmental biology Vol.29 No.2

        Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) is a well-known inducer of apoptotic cell death in many tumor cells. TRAIL is expressed in human placenta, and cytotrophoblast cells express TRAIL receptors. However, the role of TRAIL in human placentas and cytotrophoblast cells is not well understood. In this study a trophoblast cell line, JEG-3, was used as a model system to examine the effect of TRAIL on key intracellular signaling pathways involved in the control of trophoblastic cell apoptosis and survival. JEG-3 cells expressed receptors for TRAIL, death receptor (DR) 4, DR5, decoy receptor (DcR) 1 and DcR2. Recombinant human TRAIL (rhTRAIL) did not have a cytotoxic effect determined by MTT assay and did not induce apoptotic cell death determined by poly-(ADP-ribose) polymerase cleavage assay. rhTRAIL induced a rapid and transient nuclear translocation of nuclear factor-κB (NF-κB) determined by immunoblotting using nuclear protein extracts. rhTRAIL rapidly activated extracellular signal-regulated protein kinase (ERK) 1/2 as determined by immnoblotting for phospho-ERK1/2. However, c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38MAPK) and Akt (protein kinase B) were not activated by rhTRAIL. The ability of TRAIL to induce NF-κB and ERK1/2 suggests that interaction between TRAIL and its receptors may play an important role in trophoblast cell function during pregnancy.

      • KCI등재후보

        Expression of progranulin in Early and Late Gestation Human Placentas

        Hakhyun Ka 한국동물생명공학회(구 한국동물번식학회) 2006 Reproductive & developmental biology Vol.30 No.2

        Development of placenta is a complex process that is critical for the pregnancy and controlled by many factors including cytokines, hormones, growth factors and apoptotic molecules. Recently, it has been shown that progranulin (PGRN) functions in growth of embryo and trophectoderm as well as cell migration. To initiate understanding the role of PGRN in human placental development, we investigated the expression of PGRN mRNA and protein in early and late gestation human placentas, term cytotrophoblast cells and two choriocarcinoma cell lines, JEG-3 and Jar. Reverse transcriptase polymerase chain reaction identified mRNAs derived from the PGRN gene in all samples. Immunoblot analysis showed that PGRN proteins are present in early and late gestation human placentas with decreasing levels over gestation and that PGRN proteins are present in normal and transformed trophoblast cells. Immunohistochemical analysis using paraformaldehyde-fixed tissue sections taken from early and late stages of pregnancy showed that PGRN proteins are present in cytotrophoblast cells, syncytiotrophoblast and extravillous cytotrophoblast cells and that expression pattern of PGRN differed according to the stage of cell differentiation. The results of this study are consistent with the hypothesis that PGRN proteins have critical roles in placental development and suggest that PGRN may function in trophoblast cell growth and differentiation.

      • Interactions between the Maternal Uterus and the Implanting Conceptus for the Establishment of Pregnancy in Pigs

        Hakhyun Ka 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

        The successful establishment and maintenance of pregnancy is achieved by well-coordinated interactions between the maternal uterus and the implanting conceptus. In pigs, the conceptus undergoes dramatic morphological and functional changes, and secretes various biological products such as estrogens and cytokines, interleukin-1beta (IL1B), interferon-gamma (IFNG), and IFN-delta (IFND) during the implantation period. The uterine endometrium in response to the conceptus-derived molecules and ovarian progesterone becomes receptive to the conceptus by changing cell adhesion molecule expression, epithelial cell depolarization and secretory activity. Conceptus-derived estrogen acts as the maternal pregnancy recognition signal which changes the direction of endometrial prostaglandin (PG) F2 secretion from the uterine vasculature into the uterine lumen. Estrogen also induces the expression of a variety of endometrial genes, including AKR1B1, FGF7, LPAR3, and SPP1. The function of cytokines, IL1B, IFNG, and IFND, in the endometrium is not fully understood, but some recent work shows that IL1B is involved in the synthesis and transport of endometrial PGs by regulating endometrial expression of PG-synthetic enzymes, PTGS1, PTGS2, and AKR1B1, and PG transporters, ABCC4 and SLCO2A1. Estrogen and IL1B also stimulate endometrial expression of IFN signaling molecules, suggesting that estrogen and IL1B act cooperatively on priming the endometrial function of conceptus IFNG and IFND. In turn, IFNG derived from the elongating conceptuses, induces many endometrial genes, including CXCL9, CXCL10, CXCL12, and SLA-DQ. The role of IFND at the maternal-conceptus interface is not well understood yet. Further analysis of the molecules derived from the endometrium and conceptus will provide insights into the cellular and molecular basis of maternal-conceptus interactions for the establishment of successful pregnancy in pigs.

      • Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand Activates Pro-Survival Signaling Pathways, Nuclear Factor-kB and Extracellular Signal-Regulated Kinase 1/2 in Trophoblast Cell Line, JEG-3

        Ka Hakhyun 한국동물생명공학회(구 한국동물번식학회) 2005 Reproductive & Developmental Biology(Supplement) Vol.29 No.2s

        Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) is a well-known inducer of apoptotic cell death in many tumor cells. 1RAIL is expressed in human placenta, and cytotrophoblast cells express 1RAIL receptors. However, the role of TRAIL in human placentas and cytotrophoblast cells is not. well understood. In this study a trophoblast cell line, JEG-3, was used as a model system to examine the effect of TRAIL. on key intracellular signaling pathways involved in the control of trophoblastic cell apoptosis and survival JEG-3 cells expressed receptors for 1RAIL, death receptor (DR) 4, DR5, decoy receptor (OcR) 1 and DeR2. Recombinant human TRAIL (rhTRAIL) did not have a cytotoxic effect determined by MIT assay and did not induce apoptotic cell death determined by poly-(ADP-ribose) polymerase cleavage assay. rhTRAIL induced a rapid and transient nuclear translocation of nuclear factor-kB(NF-kB) determined by immunoblotting using nuclear protein extracts. rhTRAIL rapidly activated extracellular signal-regulated protein kinase (ERK) 1/2 as determined by immnoblotting for phospho-ERK1/2. However, c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38MAPK) and Akt (protein kinase B) were not activated by rhTRAIL. The ability of 1RAIL to induce NF-kB and ERK1/2 suggests that interaction between TRAIL and its receptors may play an important role in trophoblast cell function during pregnancy.

      • Endometrial response to conceptus-derived estrogen and interleukin-1β at the time of implantation in pigs

        Ka, Hakhyun,Seo, Heewon,Choi, Yohan,Yoo, Inkyu,Han, Jisoo BioMed Central 2018 Journal of animal science and biotechnology Vol.9 No.-

        <P>The establishment of pregnancy is a complex process that requires a well-coordinated interaction between the implanting conceptus and the maternal uterus. In pigs, the conceptus undergoes dramatic morphological and functional changes at the time of implantation and introduces various factors, including estrogens and cytokines, interleukin-1β2 (IL1B2), interferon-γ (IFNG), and IFN-δ (IFND), into the uterine lumen. In response to ovarian steroid hormones and conceptus-derived factors, the uterine endometrium becomes receptive to the implanting conceptus by changing its expression of cell adhesion molecules, secretory activity, and immune response. Conceptus-derived estrogens act as a signal for maternal recognition of pregnancy by changing the direction of prostaglandin (PG) F<SUB>2α</SUB> from the uterine vasculature to the uterine lumen. Estrogens also induce the expression of many endometrial genes, including genes related to growth factors, the synthesis and transport of PGs, and immunity. IL1B2, a pro-inflammatory cytokine, is produced by the elongating conceptus. The direct effect of IL1B2 on endometrial function is not fully understood. IL1B activates the expression of endometrial genes, including the genes involved in IL1B signaling and PG synthesis and transport. In addition, estrogen or IL1B stimulates endometrial expression of IFN signaling molecules, suggesting that estrogen and IL1B act cooperatively in priming the endometrial function of conceptus-produced IFNG and IFND that, in turn, modulate endometrial immune response during early pregnancy. This review addresses information about maternal-conceptus interactions with respect to endometrial gene expression in response to conceptus-derived factors, focusing on the roles of estrogen and IL1B during early pregnancy in pigs.</P>

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