• Journal of Embryo Transfer
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• v.26 no.1
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• pp.79-84
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• 2011

This study was performed to identify the differentially methylated region (DMR) and to examine the mRNA expression of the imprinted H19 gene in day 35 of SCNT pig fetuses. The fetus and placenta at day 35 of gestation fetuses after natural mating (Control) or of cloned pig by somatic cell nuclear transfer (SCNT) were isolated from a uterus. To investigate the mRNA expression and methylation patterns of H19 gene, tissues from fetal liver and placenta including endometrial and extraembryonic tissues were collected. The mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. Bisulfite analyses demonstrated that the differentially methylated region (DMR) was located between -1694 bp to -1338 bp upstream from translation start site of the H19 gene. H19 DMR (-1694 bp to -1338 bp) exhibits a normal mono allelic methylation pattern, and heavily methylated in sperm, but not in oocyte. In contrast to these finding, the analysis of the endometrium and/or extraembryonic tissues from SCNT embryos revealed a complex methylation pattern. The DNA methylation status of DMR Region In porcine H19 gene upstream was hypo methylated in SCNT tissues but hypermethylated in control tissues. Furthermore, the mRNA expression of H19 gene in liver, endometrium, and extraembryonic tissues was significantly higher in SCNT than those of control (p<0.05). These results suggest that the aberrant mRNA expression and the abnormal methylation pattern of imprinted H19 gene might be closely related to the inadequate fetal development of a cloned fetus, contributing to the low efficiency of genomic reprogramming.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.233-239
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• 2006

This study was carried out to investigate the effect of intra-uterine infusion of lipopolysaccharides (LPS) on uterine involution in dairy cows with retained placenta. The LPS isolated from Bacteroides helcogenes and Fusobaoterium varium was infused at the rate of 100 ug (n=11) and 200 ug (n=11) with 30 ml of phosphate buffer saline in each cow at 20 days post-partum. The cows in conrol group (n=11) with retained placenta were infused with 30 ml phosphate buffer saline in each cow at 20 days postpartum. Cross-sectional ultrasonography was performed at days 40 postpartum to examine uterine involution and exudate in the all dairy cows in study. The cows in control group have shown 45.5, 27.3 and 27.3% gravid horn with less than 30 mm (normal), $31{\sim}50mm$ (medium) and more than 51 mm (large) in diameter. Respective average values of gravid horn diameter in cows treated with LPS 200 ug were noticed 72.7, 18.2 and 9.1%. However, the gravid horn diameter in all the cows treated with 100 ug of LPS was less than 30 mm. The 18.2, 63.6 and 54.6% cows in the control, LPS 100 ug and LPS 200 ug, respectively, have shown no exudate in uterine cavity. However, the respective rates of small amount of exudate retention in uterine cavity were 45.5, 0.0, and 9.1%. Approximately 36.4% cows in all 3 treated groups have shown minute quantity of exudate. In conclusion, the LPS intra-uterine infusion promoted postpartum uterine involution in dairy cows with retained placenta.

• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.195-202
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• 2009

Abnormal development and fetal loss during the post-implantation period are key concerns in the production of cloned animals by somatic cell nuclear transfer (SCNT). We hypothesized that the problems in cloned porcine offspring derived from SCNT are related to interactions between the conceptus and the endometrial environment. In the present study, we investigated expression patterns in the formation of placenta-related genes (Cdx2 and GATA6) in whole in vivo normal porcine embryos (from single cell to blastocyst) and each tissue of a normal fetus at Days 25, 35 and 55 by quantitative mRNA expression analysis using real-time PCR. The expression of Cdx2 and GATA6 mRNA increased to around the blastocyst stage. These genes were gradually decreased from the peri-implantation to post-implantation stage. Moreover, we examined the expression patterns of Cdx2 and GATA6 in Day 35 normal and SCNT cloned fetuses by the same methods. And, the level of Cdx2 and GATA6 gene expression in the extraembryonic tissue of SCNT was significantly higher than that of control tissues. From the present results, it can be postulated that the aberrant expression of Cdx2 and GATA6 genes in the endometrial and extraembryonic tissues at pre- and peri-implantation stages may be closely related to the lower efficiency of animal cloning.

• Journal of Nutrition and Health
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• v.17 no.1
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• pp.50-59
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• 1984

This study was performed to investigate the placental transfer and the human fetal utilization of amino acids at term of pregnancy. The plasma levels of 23 free amino acids in both the umbilical circulation ( umbilical vein and artery ) and the uterine circulation ( uterine vein and iliac artery ) of 34 pregnant women were measured at delivery by the cesarean section. In the umbilical circulation, 9 amino acids ( alanine, lysine, valine, leucine, arginine, isoleucine, ornithine, cystine, ${\alpha}-aminobutyrate$ ) were significantly higher and 2 amino acids ( glutamate, aspartate ) were significantly lower in the umbilical vein than in the umbilical artery. In the uterine circulation, alanine, tyrosine and methionine were significantly lower in the uterine vein than in the iliac artery. Glutamate was significantly lower in the uterine vein than in the iliac artery. According to these results, the origin of fetal plasma amino acids was discussed in terms of the metabolic conversions which would occur in the Placenta and the fetal utilization of amino acids was estimated.

• Clinical and Experimental Pediatrics
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• v.50 no.3
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• pp.223-229
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• 2007

The fetus is completely dependent on mother for glucose and other nutrient transfer across the placenta. At birth, when the maternal supply is discontinued, the neonate must adjust to an independent existence. The changes in the neonate's glucose homeostasis during this transition to the extrauterine environment are influenced by the mother's metabolism and intrinsic fetal and placental problems. Maturation of carbohydrate homeostasis results from a balance between substrate availability and coordination of developing hormonal, enzymatic, and neural systems. These mechanisms may not be fully developed in neonates, so the neonate is vulnerable to carbohydrate disequilibrium resulting in damage to the central nervous system. Hypoglycemia is a relatively common metabolic problem seen during newborn care. However its definition, management and long term sequalae remain controversial. Hyporglycemia occurs frequently as a transient disorder with excellent prognosis. It also may persist and recur and cause permanent neurological complications. Although the key to effective treatment of hypoglycemia is diagnostic specific, the maintenance of euglycemia is critical to the preservation of central nervous system function. This article discusses physiology of perinatal glucose homeostasis, focusing on evaluation and treatment of hypoglycemia.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.111-117
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• 2014

This study was conducted to optimize the efficiency of cloning and to produce cloned mice. The majority of cloned mammals derived by nuclear transfer (NT) die during gestation and have enlarged and dysfunctional placentas. In this study, the optimized conditions were established to produce clone mice. The parthenogenetic oocytes were activated after 6 h regardless of cytochalasin B (CB) concentration. CB treatment ($2{\mu}g/ml$) was found second polar body. Lower concentration of CB was decreased the activation rate, but the second polar body was the best highly increased during 6 h incubation. The small fragments were exhibited in the $5{\mu}g/ml$ treatment of CB, but it was not found in lower concentration groups (> $2.5{\mu}g/ml$). To examine effects of $SrCl_2$ on the adult cumulus cells, somatic cell NT oocytes were exposed during 0.5, 1 and 6 hrs. The second polar body was significantly greater in 0.5 h exposure group (6.6%) than 1, 6 hrs. Developmental rate from 2-cell to 4-cell was the lowest in 7.5 mM Strontium chloride ($SrCl_2$) groups (84.1% and 64.3%) than 5, 10 m $MSrCl_2$. The implantation rate was not significantly difference among 5, 7.5 and 10 m $MSrCl_2$ group. Three live fetuses were produced by SCNT. SCNT placentas were remarkably heavier than IVF group (8 fetuses) (0.34, 0.34, 0.33 vs 0.14 g) compared with the placenta weight of IVF and SCNT clones.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.34-36
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• 2002

Epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) have been shown to stimulate proliferation and differentiation of various somatic cells, including placental trophoblasts and also to enhance fetal growth and development when maternally administered. Since an increase of the expression of placental EGF and IGF-I receptors in rat, mouse, and human with the gestation advanced, both EGF and IGF-I were considered to play pivotal roles on fetal growth by regulating some function of placental cells. Amino acids are crucial importance for both maternal and fetal requirements of energy source and essential constituent of fetal mass during pregnancy. Impaired fetal and placental uptake of amino acids has been observed in several models of growth retardation in the rat. Amino acid is concentrated in the fetal side through active transport by amino acid transporters and is one of the important metabolic fuels for the fatal growth. Therefore, at first plasma amino acid concentrations in mothers and fetuses were measured as an index of uphill transport across the placenta associated with EGF and IGF-1. The EGF administration at the concentration of 0, 0.1, or 0.2 $\mu\textrm{g}$/g to pregnant rats from day 18 to 21 of gestation apparently increased fetal/maternal ratio of serum proline concentration and also fatal growth in EGF dose-dependent manner. When IGF-I in doses of 0, 1, 2, and 4 $\mu\textrm{g}$/g were administrated, the ratio of leucine, isoleucine, tryptophan, phenylalanine, tyrosine and also fetal growth significantly increased with a dose-dependent manner. These results suggested that EGF and IGF-I enhanced fatal growth by, as one of its possible mechanisms, promoting placental activity to transfer some amino acid supplies from the mother to the fetus in late pregnancy.

• Journal of Photoscience
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• v.9 no.2
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• pp.472-474
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• 2002

Till date the phenomenon of maternal transfer of photic information was reported to regulate the fetal/neonatal growth, however its influence on neonatal immune system is still an enigma. In the present study, we observed an increase in maternal plasma melatonin level under short day length (SOL) condition with a consequent decrease in TLC and LC in their respective neonates. However, a significant decrease in maternal plasma melatonin level was noted under constant darkness (DD) with an increase in TLC and LC of their neonates. The blastogenic response (BGR) to Con A of splenocytes exhibited a significant increase in neonates of SDL females and a significant decrease in the neonates of DD females. Hence, it appears that the increase in maternal plasma melatonin under SOL condition transmitted information to decrease the immune status. Continuous exposure of females to darkness (DD) negatively regulated the maternal pineal gland activity thereby decreasing their plasma melatonin level. This information was transmitted for elevation of immune status in neonates, so that they exhibit better growth and sexual maturation. Therefore, we may suggest that the maternal photic information transmitted either prenatally through placenta or postnatally via the milk regulate the hormonal profile of Melatonin to regulate the immune status of neonates in order to influence their growth and sexual maturation.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.7-8
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• 2003

Since it was first reported in 1997, somatic cell cloning has been demonstrated in several other mammalian species. On the mouse, it can be cloned from embryonic stem (ES) cells, fetus-derived cells, and adult-derived cells, both male and female. While cloning efficiencies range from 0 to 20%, rates of just 1-2% are typical (i.e. one or two live offspring per one hundred initial embryos). Recently, abnormalities in mice cloned from somatic cells have been reported, such as abnormal gene expression in embryo (Boiani et al., 2001, Bortvin et al., 2003), abnormal placenta (Wakayama and Yanagimachi 1999), obesity (Tamashiro et ai, 2000, 2002) or early death (Ogonuki et al., 2002). Such abnormalities notwithstanding, success in generating cloned offspring has opened new avenues of investigation and provides a valuable tool that basic research scientists have employed to study complex processes such as genomic reprogramming, imprinting and embryonic development. On the other hand, mouse ES cell lines can also be generated from adult somatic cells via nuclear transfer. These 'ntES cells' are capable of differentiation into an extensive variety of cell types in vitro, as well assperm and oocytes in vivo. Interestingly, the establish rate of ntES cell line from cloned blastocyst is much higher than the success rate of cloned mouse. It is also possible to make cloned mice from ntES cell nuclei as donor, but this serial nuclear transfer method could not improved the cloning efficiency. Might be ntES cell has both character between ES cell and somatic cell. A number of potential agricultural and clinical applications are also are being explored, including the reproductive cloning of farm animals and therapeutic cloning for human cell, tissue, and organ replacement. This talk seeks to describe both the relationship between nucleus donor cell type and cloning success rate, and methods for establishing ntES cell lines. (중략)

• Journal of Nutrition and Health
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• v.18 no.3
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• pp.209-216
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• 1985

The plasma levels of 23 free amino acids in both the umbilical circulation (umbilical vein and artery) and the maternal circulation (antecubital vein, uterine vein and iliac artery ) of 34 pregnant women were measured at delivery by the cesarean section. Each amino acid with the exception of glutamate was found to be in higher concentration in the fetal blood and the cord plasma characterized by hyperaminoacidemia with a marked increase of the lysine and ornithine levels. The linear relationships found between the amino acids concentrations of iliac artery and those of umbilical vein suggest three transport groups across the placental membrane One group consists of neutral amino acids whose slopes are equal to one and the other two groups are characterized by their slopes higher and lower than unity respectivly. This division into three groups is tentatively explained by the result of a dynamic equilibrium between active transport towards the fetus and diffusion back towards the maternal circulation.