Th 17 Cells and Nesfatin-1 are associated with Spontaneous Abortion in the CBA/j×DBA/2 Mouse Model

Chung, Yiwa,Kim, Heejeong,Im, Eunji,Kim, Philjae,Yang, Hyunwon The Korean Society of Developmental Biology 2015 발생과 생식 Vol.19 No.4

The pregnancy and abortion process involves a complex mechanism with various immune cells present in the implantation sites and several hormones associated with pregnancy, such as leptin, ghrelin and nesfatin-1. However, the mechanism underlying spontaneous abortion by maternal T helper 17 (Th17) present in the implantation sites and nesfatin-1, which is of anorexigenic hormones, is not fully understood so far. Therefore, the purpose of this study was to examine the possible roles of Th17 cells present in the implantation sites and nesfatin-1 expressed in the uterus on spontaneous abortion using the $CBA/j{\times}DBA/2$ mouse model. Th17 transcription factor, ROR-${\gamma}t$ mRNA expression was significantly increased in the abortion sites compared with the implantation sites of abortion model mice on day 14.5 and 19.5 of pregnancy. In addition, the expression levels of IL-17A mRNA were significantly higher in abortion sites than in implantation sites on day 14.5 and 19.5. Moreover, the nesfatin-1/NUCB2 protein and mRNA levels were increased in abortion sites compared with levels in implantation sites of both normal pregnant and abortion model mice on day 14.5 of pregnancy. Interestingly, nesfatin-1/NUCB2 serum levels were not changed throughout the whole pregnancy in abortion model mice, but its serum level was dramatically increased on day 14.5, and then rapidly decreased on day 19.5 in normal pregnant mice. In this study, we showed for the first time the expression of nesfatin-1/NUCB2 mRNA and protein in implantation sites during pregnancy. The present results suggest that Th17 cells in the uterus may play an important role in the period of implantation and for maintenance of pregnancy. Furthermore, the present results suggest that Th17 cells in implantation sites may be a key regulator for maintenance of pregnancy and provides evidence that activation of these cells may be regulated by nesfatin-1/NUCB2. Further study is needed to elucidate the role of nesfatin-1 expressed in the uterus during pregnancy.

Expression of Nesfatin-1/NUCB2 in Fetal, Neonatal and Adult Mice

Yiwa Chung,Eunhye Jung,Heejung Kim,Jinhee Kim,양현원 한국발생생물학회 2013 발생과 생식 Vol.17 No.4

Nesfatin-1/NUCB2, which is associated with the control of appetite and energy metabolism, was reported for the first time to be expressed in the hypothalamus. However, recent studies have shown that nesfatin-1/NUCB2 was expressed not only in the hypothalamus, but also in various tissues including digestive and reproductive organs. We also demonstrated that nesfatin-1/NUCB2 was expressed in the reproductive organs, pituitary gland, heart, lung, and gastrointestinal tract of the adult mouse. However, little is known about nesfatin-1/NUCB2 expression in fetal and neonatal mice. Therefore, we examined here the distribution of nesfatin-1/NUCB2 in various organs of fetal and neonatal mice and compared them with the distribution in adult mice. As a result of immunohistochemical staining, nesfatin-1/NUCB2 protein was expressed relatively higher in the lung, kidney, heart, and liver compared to other organs in the fetus. Western blot results also showed that nesfatin-1/NUCB2 protein was detected in the lung, kidney, heart, and stomach. Next, we compared the expression levels of nesfatin-1/NUCB2 mRNA in the fetus and neonate with the expression levels in both male and female adult mice. The expression levels in heart, lung, stomach, and kidney were higher compared with other organs in fetal and neonatal mice and in both male and female adult mice. Interestingly, the expression of nesfatin-1/NUCB2 mRNA in the kidney was dramatically increased in male and female adult mice compared to fetal and neonatal mice. These results indicate that nesfatin-1/NUCB2 may regulate the development and physiological function of mouse organs. In the future, we need more study on the function of nesfatin-1/NUCB2, which is highly expressed in the heart, lung, and kidney during mouse development.

Expression of Nesfatin-1/NUCB2 in Fetal, Neonatal and Adult Mice

Chung, Yiwa,Jung, Eunhye,Kim, Heejung,Kim, Jinhee,Yang, Hyunwon The Korean Society of Developmental Biology 2013 발생과 생식 Vol.17 No.4

Nesfatin-1/NUCB2, which is associated with the control of appetite and energy metabolism, was reported for the first time to be expressed in the hypothalamus. However, recent studies have shown that nesfatin-1/NUCB2 was expressed not only in the hypothalamus, but also in various tissues including digestive and reproductive organs. We also demonstrated that nesfatin-1/NUCB2 was expressed in the reproductive organs, pituitary gland, heart, lung, and gastrointestinal tract of the adult mouse. However, little is known about nesfatin-1/NUCB2 expression in fetal and neonatal mice. Therefore, we examined here the distribution of nesfatin-1/NUCB2 in various organs of fetal and neonatal mice and compared them with the distribution in adult mice. As a result of immunohistochemical staining, nesfatin-1/NUCB2 protein was expressed relatively higher in the lung, kidney, heart, and liver compared to other organs in the fetus. Western blot results also showed that nesfatin-1/NUCB2 protein was detected in the lung, kidney, heart, and stomach. Next, we compared the expression levels of nesfatin-1/NUCB2 mRNA in the fetus and neonate with the expression levels in both male and female adult mice. The expression levels in heart, lung, stomach, and kidney were higher compared with other organs in fetal and neonatal mice and in both male and female adult mice. Interestingly, the expression of nesfatin-1/NUCB2 mRNA in the kidney was dramatically increased in male and female adult mice compared to fetal and neonatal mice. These results indicate that nesfatin-1/NUCB2 may regulate the development and physiological function of mouse organs. In the future, we need more study on the function of nesfatin-1/NUCB2, which is highly expressed in the heart, lung, and kidney during mouse development.

Th 17 Cells and Nesfatin-1 are associated with Spontaneous Abortion in the CBA/j × DBA/2 Mouse Model

Yiwa Chung,Heejeong Kim,Eunji Im,Philjae Kim,Hyunwon Yang 한국발생생물학회 2015 발생과 생식 Vol.19 No.4

The pregnancy and abortion process involves a complex mechanism with various immune cells present in the implantation sites and several hormones associated with pregnancy, such as leptin, ghrelin and nesfatin-1. However, the mechanism underlying spontaneous abortion by maternal T helper 17 (Th17) present in the implantation sites and nesfatin-1, which is of anorexigenic hormones, is not fully understood so far. Therefore, the purpose of this study was to examine the possible roles of Th17 cells present in the implantation sites and nesfatin-1 expressed in the uterus on spontaneous abortion using the CBA/j × DBA/2 mouse model. Th17 transcription factor, ROR-γt mRNA expression was significantly increased in the abortion sites compared with the implantation sites of abortion model mice on day 14.5 and 19.5 of pregnancy. In addition, the expression levels of IL-17A mRNA were significantly higher in abortion sites than in implantation sites on day 14.5 and 19.5. Moreover, the nesfatin-1/NUCB2 protein and mRNA levels were increased in abortion sites compared with levels in implantation sites of both normal pregnant and abortion model mice on day 14.5 of pregnancy. Interestingly, nesfatin- 1/NUCB2 serum levels were not changed throughout the whole pregnancy in abortion model mice, but its serum level was dramatically increased on day 14.5, and then rapidly decreased on day 19.5 in normal pregnant mice. In this study, we showed for the first time the expression of nesfatin-1/NUCB2 mRNA and protein in implantation sites during pregnancy. The present results suggest that Th17 cells in the uterus may play an important role in the period of implantation and for maintenance of pregnancy. Furthermore, the present results suggest that Th17 cells in implantation sites may be a key regulator for maintenance of pregnancy and provides evidence that activation of these cells may be regulated by nesfatin-1/NUCB2. Further study is needed to elucidate the role of nesfatin-1 expressed in the uterus during pregnancy.

Serum Cytokine Levels are related to Nesfatin-1/NUCB2 Expression in the Implantation Sites of Spontaneous Abortion Model of CBA/j × DBA/2 Mice

Yiwa Chung,김희정,선소정,양현원 한국발생생물학회 2017 발생과 생식 Vol.21 No.1

The process of spontaneous abortion involves a complex mechanism with various cytokines, growth factors, and hormones during the pregnancy. However, the mechanism underlying spontaneous abortion by pro- and antiinflammatory cytokines in the serum during the pregnancy is not fully understood. Therefore, the purpose of this study was to examine the relationship between the serum levels of pro- and anti-inflammatory cytokines and spontaneous abortion using the CBA/j × DBA/2 mouse model. Serum levels of pro-inflammatory cytokines, such as IFN-γ, IL-1α and TNF-α were not increased in abortion model mice, but anti-inflammatory cytokines, such as IL-4, IL-13 and IL-1ra were decreased compared to normal pregnant mice. In addition, serum levels of chemokine, such as SDF-1, G-CSF, M-CSF, IL-16, KC and MCP-1 were decreased in abortion model mice compared to normal pregnant mice. However, the expression levels of nesfatin-1/NUCB2 mRNA and protein in the uteri of implantation sites were significantly higher in abortion model mice than normal pregnant mice. These results suggest that uterine nesfatin-1/NUCB2 expression may be down-regulated by inflammatory cytokines and chemokines in the serum of pregnant mice. Moreover, this study suggests the possibility that nesfatin-1/NUCB2 expressed in the implantation sites may be associated with the maintenance of pregnancy.

Serum Cytokine Levels are related to Nesfatin-1/NUCB2 Expression in the Implantation Sites of Spontaneous Abortion Model of CBA/j×DBA/2 Mice

Chung, Yiwa,Kim, Heejeong,Seon, Sojeong,Yang, Hyunwon The Korean Society of Developmental Biology 2017 발생과 생식 Vol.21 No.1

The process of spontaneous abortion involves a complex mechanism with various cytokines, growth factors, and hormones during the pregnancy. However, the mechanism underlying spontaneous abortion by pro- and anti-inflammatory cytokines in the serum during the pregnancy is not fully understood. Therefore, the purpose of this study was to examine the relationship between the serum levels of pro- and anti-inflammatory cytokines and spontaneous abortion using the $CBA/j{\times}DBA/2$ mouse model. Serum levels of pro-inflammatory cytokines, such as $IFN-{\gamma}$, $IL-1{\alpha}$ and $TNF-{\alpha}$ were not increased in abortion model mice, but anti-inflammatory cytokines, such as IL-4, IL-13 and IL-1ra were decreased compared to normal pregnant mice. In addition, serum levels of chemokine, such as SDF-1, G-CSF, M-CSF, IL-16, KC and MCP-1 were decreased in abortion model mice compared to normal pregnant mice. However, the expression levels of nesfatin-1/NUCB2 mRNA and protein in the uteri of implantation sites were significantly higher in abortion model mice than normal pregnant mice. These results suggest that uterine nesfatin-1/NUCB2 expression may be down-regulated by inflammatory cytokines and chemokines in the serum of pregnant mice. Moreover, this study suggests the possibility that nesfatin-1/NUCB2 expressed in the implantation sites may be associated with the maintenance of pregnancy.

Th 17 Cells and Nesfatin-1 are associated with Spontaneous Abortion in the CBA/j × DBA/2 Mouse Model

Yiwa Chung,Heejeong Kim,Eunji Im,Philjae Kim,양현원 한국발생생물학회 2015 발생과 생식 Vol.19 No.4

The pregnancy and abortion process involves a complex mechanism with various immune cells present in the implantation sites and several hormones associated with pregnancy, such as leptin, ghrelin and nesfatin-1. However, the mechanism underlying spontaneous abortion by maternal T helper 17 (Th17) present in the implantation sites and nesfatin-1, which is of anorexigenic hormones, is not fully understood so far. Therefore, the purpose of this study was to examine the possible roles of Th17 cells present in the implantation sites and nesfatin-1 expressed in the uterus on spontaneous abortion using the CBA/j × DBA/2 mouse model. Th17 transcription factor, ROR-γt mRNA expression was significantly increased in the abortion sites compared with the implantation sites of abortion model mice on day 14.5 and 19.5 of pregnancy. In addition, the expression levels of IL-17A mRNA were significantly higher in abortion sites than in implantation sites on day 14.5 and 19.5. Moreover, the nesfatin-1/NUCB2 protein and mRNA levels were increased in abortion sites compared with levels in implantation sites of both normal pregnant and abortion model mice on day 14.5 of pregnancy. Interestingly, nesfatin- 1/NUCB2 serum levels were not changed throughout the whole pregnancy in abortion model mice, but its serum level was dramatically increased on day 14.5, and then rapidly decreased on day 19.5 in normal pregnant mice. In this study, we showed for the first time the expression of nesfatin-1/NUCB2 mRNA and protein in implantation sites during pregnancy. The present results suggest that Th17 cells in the uterus may play an important role in the period of implantation and for maintenance of pregnancy. Furthermore, the present results suggest that Th17 cells in implantation sites may be a key regulator for maintenance of pregnancy and provides evidence that activation of these cells may be regulated by nesfatin-1/NUCB2. Further study is needed to elucidate the role of nesfatin-1 expressed in the uterus during pregnancy.

The Tissue Distribution of Nesfatin-1/NUCB2 in Mouse

Jinhee Kim,Yiwa Chung,Heejeong Kim,Eunji Im,Hyojin Lee,Hyunwon Yang 한국발생생물학회 2014 발생과 생식 Vol.18 No.4

Nesfatin-1, an anorexic nucleobindin-2 (NUCB2)-derived hypothalamic peptide, controls appetite and energy metabolism. Recent studies show that nesfatin-1/NUCB2 is expressed not only in the brain but also in gastric and adipose tissues. Thus, we investigated the distributions of nesfatin-1/NUCB2 in various tissues of male and female mice by real-time PCR, western blotting, and immunohistochemical staining. Real-time PCR analyses showed that NUCB2 mRNA was predominantly expressed in the pituitary and at lower levels in the hypothalamus, spleen, thymus, heart, liver, and muscle of both male and female mice. Expression was much higher in reproductive organs, such as the testis, epididymis, ovary, and uterus, than in the hypothalamus. Western blot analysis of the nesfatin-1 protein level showed similar results to the real-time PCR analyses in both male and female mice. These results suggest that nesfatin-1/NUCB2 have widespread physiological effects in endocrine and non-endocrine organs. In addition, immunohistochemical staining revealed that nesfatin-1 was localized in interstitial cells, including Leydig cells and in the columnar epithelium of the epididymis. Nesfatin-1 was also expressed in theca cells and interstitial cells in the ovary and in epithelial cells of the endometrium and uterine glands in the uterus. These results suggest that nesfatin-1 is a novel potent regulator of steroidogenesis and gonadal function in male and female reproductive organs. Further studies are required to elucidate the functions of nesfatin-1 in various organs of male and female mice.

The Tissue Distribution of Nesfatin-1/NUCB2 in Mouse

Kim, Jinhee,Chung, Yiwa,Kim, Heejeong,Im, Eunji,Lee, Hyojin,Yang, Hyunwon The Korean Society of Developmental Biology 2014 발생과 생식 Vol.18 No.4

Nesfatin-1, an anorexic nucleobindin-2 (NUCB2)-derived hypothalamic peptide, controls appetite and energy metabolism. Recent studies show that nesfatin-1/NUCB2 is expressed not only in the brain but also in gastric and adipose tissues. Thus, we investigated the distributions of nesfatin-1/NUCB2 in various tissues of male and female mice by real-time PCR, western blotting, and immunohistochemical staining. Real-time PCR analyses showed that NUCB2 mRNA was predominantly expressed in the pituitary and at lower levels in the hypothalamus, spleen, thymus, heart, liver, and muscle of both male and female mice. Expression was much higher in reproductive organs, such as the testis, epididymis, ovary, and uterus, than in the hypothalamus. Western blot analysis of the nesfatin-1 protein level showed similar results to the real-time PCR analyses in both male and female mice. These results suggest that nesfatin-1/NUCB2 have widespread physiological effects in endocrine and non-endocrine organs. In addition, immunohistochemical staining revealed that nesfatin-1 was localized in interstitial cells, including Leydig cells and in the columnar epithelium of the epididymis. Nesfatin-1 was also expressed in theca cells and interstitial cells in the ovary and in epithelial cells of the endometrium and uterine glands in the uterus. These results suggest that nesfatin-1 is a novel potent regulator of steroidogenesis and gonadal function in male and female reproductive organs. Further studies are required to elucidate the functions of nesfatin-1 in various organs of male and female mice.

Thymocyte Differentiation is Regulated by a Change in Estradiol Levels during the Estrous Cycle in Mouse

Lee, Hyojin,Kim, Heejung,Chung, Yiwa,Kim, Jinhee,Yang, Hyunwon The Korean Society of Developmental Biology 2013 발생과 생식 Vol.17 No.4

Recent study showed that T cells in the immune organs and peripheral blood are influenced by estradiol, leading to a dysfunction of the immune system. However, little is known about the thymic-gonadal relationship during the estrous cycle in mouse. Therefore, the purpose of this study was to elucidate the mechanism by which a change in estradiol levels during the estrous cycle regulates the development of T cells in the mouse thymus. Six-week-old ICR mice were used and divided into four groups, including diestrous, proestrous, estrous, and metestrous. We first confirmed that ER-${\alpha}$ and - ${\beta}$ estrogen receptors were expressed in thymic epithelial cells, showing that their expression was not different during the estrous cycle. There was also no significant difference in thymic weight and total number of thymocytes during the estrous cycle. To determine the degree of thymocyte differentiation during the estrous cycle, we analyzed thymocytes by flow cytometry. As a result, the percentage of CD4+CD8+ double-positive (DP) T cells was significantly decreased in the proestrous phase compared to the diestrous phase. However, CD4+CD8- or CD4-CD8+ (SP) T cells were significantly increased in the proestrous phase compared to the diestrous phase. In addition, the percentage of CD44+CD25- (DN1) T cells was significantly decreased in the estrous phase compared to other phases, whereas the percentages of CD44+CD25+ (DN2), CD44-CD25+ (DN3), and CD44-CD25- (DN4) were not changed during the estrous cycle. These results indicate that the development of thymocytes may arrest in the DP to SP transition stage in the proestrous phase displaying the highest serum level of estradiol. This study suggests that a change in estradiol levels during the estrous cycle may be involved in the regulation of thymocyte differentiation in the mouse thymus.