자궁 내 배아 착상 환경에 대한 이해

송행석,Song, Haengseok 대한생식의학회 2006 Clinical and Experimental Reproductive Medicine Vol.33 No.3

Etv5, a transcription factor with versatile functions in male reproduction

Eo, Jinwon,Song, Haengseok,Lim, Hyunjung Jade The Korean Society for Reproductive Medicine 2012 Clinical and Experimental Reproductive Medicine Vol.39 No.2

Transcription factors govern diverse aspects of cell growth and differentiation as major switches of gene expression. Etv5, a member of the E26 transformation-specific family of transcription factors, has many stories to share when it comes to reproduction. Etv5 deficient mice show complex infertility phenotypes both in males and females. In males, the infertility phenotype exhibited by Etv5 deficiency is sexually dimorphic, and it involves both somatic cells and germ cells. In $Etv5^{-/-}$ female mice, the problem is more complicated by hormonal involvement. This review synthesizes old and new information on this versatile transcription factor-from the inadvertent discovery of its role in the testes to its newly discovered role in maintaining spermatogonial stem cells.

Regulation of Differentiation Potential of Human Mesenchymal Stem Cells by Intracytoplasmic Delivery of Coactivator-Associated Arginine Methyltransferase 1 Protein Using Cell-Penetrating Peptide

Jo, Junghyun,Song, Haengseok,Park, Sang Gyu,Lee, Soo-Hong,Ko, Jung-Jae,Park, Jong-Hyuk,Jeong, Jaemin,Cheon, Yong-Pil,Lee, Dong Ryul Wiley (John WileySons) 2012 Stem cells Vol.30 No.8

<P>Recent studies suggest that epigenetic modifications, such as DNA methylation and histone modification, can alter the differentiation potential of stem cells or progenitor cells. Specifically, coactivator-associated arginine methyltransferase 1 (CARM1) is known to act as a coactivator for various transcription factors and to regulate gene expression by chromatin remodeling through histone methylation. Here, for the first time, we have used direct protein delivery of CARM1 using cell-penetrating peptide (CPP) to regulate the differentiation potential of human mesenchymal stem cells (hMSCs). Immunofluorescence showed that the CPP-CARM1 protein is successfully delivered into the nuclei of hMSCs. Further experiments using immunofluorescence and Western blotting showed that the delivered CARM1 protein can effectively methylate the arginine 17 residue of histone H3 in both bone marrow (BM)- and adipose-derived (AD)-hMSCs, thus suggesting that the CARM1 protein delivered by the CPP system is biologically active in hMSCs. Chromatin immunoprecipitation (ChIP) assay and genome-wide gene expression profiling supported the result that delivered CARM1 protein can cause chromatin remodeling through histone methylation. Finally, the CPP-CARM1 protein efficiently elevated the differentiation efficiency of BM-hMSCs and AD-hMSCs into adipogenic, osteogenic, and myogenic cell lineages in vitro. Altered expression of critical genes after hMSC differentiation was reconfirmed by real-time reverse transcription polymerase chain reaction (qRT-PCR). Collectively, our results suggest that CPP-CARM1 can elevate the differentiation potential of hMSCs into various cell types, and that this system using CPP is a useful tool for exogenous protein delivery in clinical applications of cell-based therapy.</P>

Melatonin significantly down-regulates long non-coding RNA Cox2 in M1 macrophages stimulated with LPS

Dohyeong Kim,Haengseok Song 한국실험동물학회 2022 한국실험동물학회 학술발표대회 논문집 Vol.2022 No.7

Autophagic activation in vitrified–warmed mouse oocytes

Bang, Soyoung,Shin, Hyejin,Song, Haengseok,Suh, Chang Suk,Lim, Hyunjung Jade BioScientifica 2014 Reproduction Vol.148 No.1

<P>Vitrification involves the use of cryoprotectants (CPAs) and liquid nitrogen (LN<SUB>2</SUB>), which may cause osmotic damage and cryoinjury to oocytes. Autophagy is widely recognized as a survival or response mechanism elicited by various environmental and cellular stressors. However, the induction of autophagy in vitrified–warmed oocytes has not been examined. In this work, we investigated whether the vitrification–warming process induces autophagy in mouse oocytes. Metaphase II (MII) oocytes that were vitrified and stored in LN<SUB>2</SUB> for at least 2 weeks were used in the study. In RT-PCR analyses, we observed that several <I>Atg</I> genes such as <I>Atg5</I>, <I>Atg7</I>, <I>Atg12</I>, <I>LC3a</I> (<I>Map1lc3a</I>), <I>LC3b</I> (<I>Map1lc3b</I>), and <I>Beclin1</I> were expressed in MII mouse oocytes. Slight reduction in mRNA levels of <I>Atg7</I> and <I>Atg12</I> in vitrified–warmed oocytes was noted, and expression of these genes was not significantly influenced. Confocal live imaging analysis using oocytes from GFP-LC3 transgenic mice revealed that vitrified–warmed oocytes had a significantly higher number of GFP-LC3 puncta in comparison to fresh oocytes. The expression of BECLIN1 protein was also increased in vitrified–warmed oocytes. Treatment with 3-methyladenine, an inhibitor of autophagy, did not significantly affect the rates of oocyte survival, IVF, and embryonic development after warming and IVF. The results suggest that the observed autophagic activation in vitrified–warmed oocytes is a natural adaptive response to cold stress. Collectively, we show for the first time that vitrified–warmed mouse oocytes exhibit autophagic activation during warming and that this response is not induced by CPA-containing solutions. The induction of autophagy by cold temperature is first reported herein.</P>

Suppression of autophagic activation in the mouse uterus by estrogen and progesterone

Choi, Soyoung,Shin, Hyejin,Song, Haengseok,Lim, Hyunjung Jade Society for Endocrinology 2014 The Journal of endocrinology Vol.221 No.1

<P>Autophagy is a major cellular catabolic pathway tightly associated with cell survival. The involvement of autophagy in the prolonged survival of blastocysts in the uterus is well established, and it was assumed that ovarian steroid hormones – progesterone (P<SUB>4</SUB>) and estrogens – have important roles in the regulation of autophagy. However, information is scarce regarding whether these hormones regulate autophagy in certain hormone-responsive cellular systems. In this study, we investigated the effects of estrogen and P<SUB>4</SUB> on autophagic response in the uteri of pregnant mice and in ovariectomized (OVX) mice treated with hormones. During pregnancy, autophagic response is high on days 1 and 2 when the uterus shows an inflammatory response to mating, but it subsides around the time of implantation. Dexamethasone treatment to day 1 pregnant mice reduced autophagy in the uterus. In OVX mouse uteri, estrogen or P<SUB>4</SUB> reduces autophagic response within 6 h. Glycogen content in OVX uteri was increased by 3-methyladenine treatment, suggesting that autophagy is involved in glycogen breakdown in the hormone-deprived uterus. The classical nuclear receptor antagonists, ICI 182 780 or mifepristone, lead to the recovery of the autophagic response in OVX uteri. The suppression of autophagy by 17β-estradiol is inversely correlated with the accumulation of phospho-mouse target of rapamycin, and rapamycin treatment is moderately effective in the upregulation of autophagic response in OVX mouse uteri. Collectively, this study establishes that the uterine autophagy is induced in hormone-derived environment and is suppressed by hormone treatment. Uterine autophagy may have multiple functions as a responsive mechanism to acute inflammation and as an energy provider by breaking down glycogen under hormone deprivation.</P>

Gestational Diabetes Affects the Growth and Functions of Perivascular Stem Cells

An, Borim,Kim, Eunbi,Song, Haengseok,Ha, Kwon-Soo,Han, Eun-Taek,Park, Won Sun,Ahn, Tae Gyu,Yang, Se-Ran,Na, Sunghun,Hong, Seok-Ho Korean Society for Molecular and Cellular Biology 2017 Molecules and cells Vol.40 No.6

Gestational diabetes mellitus (GDM), one of the common metabolic disorders of pregnancy, leads to functional alterations in various cells including stem cells as well as some abnormalities in fetal development. Perivascular stem cells (PVCs) have gained more attention in recent years, for the treatment of various diseases. However, the effect of GDM on PVC function has not been investigated. In our study, we isolated PVCs from umbilical cord of normal pregnant women and GDM patients and compared their phenotypes and function. There is no significant difference in phenotypic expression, response to bFGF exposure and adipogenic differentiation capacity between normal (N)-PVCs and GDM-PVCs. However, when compared with N-PVCs, early passage GDM-PVCs displayed decreased initial rates of cell yield and proliferation as well as a reduced ability to promote wound closure. These results suggest that maternal metabolic dysregulation during gestation can alter the function of endogenous multipotent stem cells, which may impact their therapeutic effectiveness.

Transcriptional profiling with a pathway-oriented analysis identifies dysregulated molecular phenotypes in the endometrium of patients with polycystic ovary syndrome.

Kim, Jin Yeong,Song, Haengseok,Kim, Hyunjoo,Kang, Hee Jung,Jun, Jin Hyun,Hong, Sung Ran,Koong, Mi Kyoung,Kim, In Sun Issued for the Endocrine Society by the Williams W 2009 The Journal of clinical endocrinology & metabolism Vol.94 No.4

<P>CONTEXT: Polycystic ovary syndrome (PCOS) is a common endocrine disorder characterized by chronic oligo/anovulation, hyperandrogenemia, infertility, and metabolic alterations related to insulin resistance. These abnormalities in PCOS may have complex effects on pathophysiology of the endometrium, contributing to infertility and endometrial disorders. OBJECTIVE: The objective of this study was to examine dysregulated signaling pathways in the endometrium of patients with PCOS (PCOSE) by analyzing expression profiles with a pathway-oriented method. DESIGN: Microarrays, RT-PCR, laser capture microdissection, and immunohistochemistry were performed with endometrial tissues. SETTING: This study was performed at a university hospital laboratory. Patients: This study comprised 12 regularly cycling women and 12 PCOS patients. MAIN OUTCOME MEASURE: Dysregulated signaling pathways in PCOSE were identified as a gene set. RESULTS: Hierarchical clustering revealed distinct expression profiles for PCOSE and the endometrium of normal cycling women. Gene sets associated with androgen signaling were not enriched in PCOSE, although they affect ovarian physiology of PCOS patients. Several biological pathways including cell cycle, apoptosis, glycolysis, and integrin-Rho-cytoskeleton network were aberrantly down-regulated in PCOSE. Expression of genes constituting these gene sets enriched in normal cycling women was systemically down-regulated in PCOSE. Laser capture microdissection-coupled real-time RT-PCR and immunohistochemistry further demonstrated that cell proliferation in the stroma, but not the epithelium, is significantly reduced in PCOSE. CONCLUSIONS: Systemic down-regulation of various signaling pathways in PCOSE with extremely prolonged proliferative phase provides insight into the abnormal phenotypes that reflect pathophysiology of PCOS in the endometrium, possibly leading to increased risks of endometrial disorders.</P>

Egr3 exhibits a unique subcellular localization in mouse oocytes during meiotic maturation

Hyejin Shin,Hyunjung Lim,Haengseok Song 한국발생생물학회 2011 한국발생생물학회 학술발표대회 Vol.30 No.-

The Egr family of zinc finger transcription factors consisting of 4 members (Egr1 to -4) regulates critical genetic programs involved in cellular growth, differentiation, and function. Especially, the critical role for Egr1 in regulating luteinizing hormone responsiveness was demonstrated by using gene-targeted mouse models. Other members of Egr family were shown to be involved in other cellular and developmental processes. To understand if Egr3 is implicated in ovarian functions, we focused on identifying cell type-specific and subcellular localization of Egr3 in cycling mouse ovaries and oocytes. RT-PCR analyses show that Egr3 mRNA is expressed in the mouse ovary and oocytes. By immunofluorescence staining, we observed that Egr3 is weakly expressed in subsets of granulosa cells. Interestingly, Egr3 seems to be co-localized with meiotic spindle in some oocytes in the ovarian section. Therefore, we examined Egr3 localization in MI oocytes cultured in vitro. We confirmed co-localization of Egr3 and microtubule in the mouse oocyte during meiosis I. Egr3 localization is noted around condensing chromosomes during prometaphase I (PMI). At metaphase I (MI) and MII, Egr3 is localized on meiotic spindle and also around each cytosolic microtubule organizing centers (MTOCs) in a punctate pattern. To examine if microtubule is required for correct positioning of Egr3 on this structure, we observed the pattern of Egr3 in oocytes matured under taxol or nocodazole. In taxol-treated oocyte, Egr3 and gamma-tubulin complex are enlarged. In nocodazole-treated oocyte, Egr3 localization on spindle and MTOCs are abolished. Thus, Egr3 localization seems to require the presence of intact microtubule. Collectively, our result shows for the first time that Egr3, a transcription factor, is localized on meiotic spindle of maturing mouse oocytes. The work suggests a novel role for Egr3 as a factor involved in MTOC dynamics during meiosis.

Egr-1 is necessary for fibroblast growth factor-2-induced transcriptional activation of the glial cell line-derived neurotrophic factor in murine astrocytes.

Shin, Soon Young,Song, Haengseok,Kim, Chang Gun,Choi, Yang-Kyu,Lee, Kyoung Sun,Lee, Seung-Jae,Lee, He-Jin,Lim, Yoongho,Lee, Young Han American Society for Biochemistry and Molecular Bi 2009 The Journal of biological chemistry Vol.284 No.44

<P>Glial cell line-derived neurotrophic factor (Gdnf) promotes neurite outgrowth and survival of neuronal cells, but its transcriptional regulation is poorly understood. Here, we sought to investigate the mechanism underlying fibroblast growth factor-2 (FGF2) induction of Gdnf expression in astrocytes. We found that FGF2 stimulation of rat astrocytes induced expression of Egr-1 at a high level. Sequence analysis of the rat Gdnf gene identified three overlapping Egr-1-binding sites between positions -185 and -163 of the rat Gdnf promoter. Transfection studies using a series of deleted Gdnf promoters revealed that these Egr-1-binding sites are required for maximal activation of the Gdnf promoter by FGF2. Chromatin immunoprecipitation analysis indicated that Egr-1 binds to the Gdnf promoter. Furthermore, the induction of Gdnf expression by FGF2 is strongly attenuated both in C6 glioma cells stably expressing Egr-1-specific small interfering RNA and in primary cultured astrocytes from the Egr-1 knock-out mouse. Additionally, we found that stimulation of the ERK and JNK pathways by FGF2 is functionally linked to Gdnf expression through the induction of Egr-1. These data demonstrate that FGF2-induced Gdnf expression is mediated by the induction of Egr-1 through activation of the ERK and JNK/Elk-1 signaling pathways.</P>