• Journal of Animal Science and Technology
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• v.52 no.3
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• pp.175-186
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• 2010

The aim of this study was to analyze the proteome expressions of proliferating stromal vascular cells from Hanwoo omental, subcutaneous and intramuscular depots subjected to hormone deprivation and IGF-1, Estradiol-$17{\beta}$ addition. For hormone deprivation or addition studies, the cells were either grown in 10% charcoal-dextran stripped fetal bovine serum (CD-FBS) or in 10% FBS supplemented medium. Further, to analyze the effect of insulin like growth factor (IGF-1) and $17\beta$-Estradiol (E2), cells were grown in 10% CD-FBS containing IGF-1 (10 ng/ml) or E2 (10 nM). The results showed that hormone deprivation had a negative impact on proliferation among the cells from all depots without any growth difference. On comparison of proliferation levels, higher levels were observed in cells that were grown in 10% FBS than in 10% CD-FBS alone or with IGF-1/E2. Proteome expression from preadipocytes grown in hormone deprivation conditions were compared by 2D-DIGE and MALDIToF/ToF. A total of twelve different proteins were found to be differentially expressed under hormone deprivation conditions. Further, our proteomic analysis with DIGE under IGF-1 and E2 addition revealed four proteins with differential expression levels. Moreover, the results highlighted in this study offer a role for each differentially expressed protein with respect to their effect in positive or negative regulation on proliferation.

• Journal of Life Science
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• v.24 no.2
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• pp.173-180
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• 2014

Estrogen can promote or inhibit cellular proliferation depending on tissue cell types and physiological condition and acts through the signal transduction pathways mediated primarily by estrogen receptors. This study examined the effects of fulvestrant (Ful), a well-known antagonist for the estrogen receptor, on the action of $17{\beta}$-estradiol (E2) with respect to the proliferation and apoptosis of Chinese hamster ovarian (CHO) cells. We used different concentrations of E2, Ful, and E2 plus Ful during different treatment durations. Treatment with 15-40 ${\mu}M$ E2 significantly inhibited proliferation in a time-dependent manner, although it had no influence in concentrations up to 1 ${\mu}M$. Interestingly, Ful at 10-40 ${\mu}M$ also inhibited cellular proliferation in both a concentration- and time-dependent manner. In addition, Ful enhanced rather than decreased the inhibitory effect on cellular proliferation by E2 in combined treatment for 10 days. Thus, Ful does not appear to have an antagonistic effect on estrogen's anti-proliferative action in CHO cells. In TUNEL assays to confirm DNA fragmentation by E2 and/or Ful, CHO cells treated with 20 ${\mu}M$ E2 showed a TUNEL-positive reaction in most DAPI-stained nuclei, and cells treated with either 40 ${\mu}M$ Ful or 40 ${\mu}M$ Ful plus 20 ${\mu}M$ E2 also exhibited a TUNEL-positive reaction but at a lower rate compared to the E2-treated cells. These results indicate that Ful does not have an antagonistic effect on estrogen's anti-proliferative action in CHO cells, suggesting that the anti-proliferative and apoptosis-related mechanism(s) through DNA fragmentation by E2 and Ful may be mediated by different signal transduction pathways.

• Journal of Life Science
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• v.23 no.12
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• pp.1454-1459
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• 2013

This study investigated the effects that water temperature and the administration of estradiol-17${\beta}$ (E2) had on the sex ratio and growth of the Japanese eel, Anguilla japonica. Glass eels (total length${\fallingdotseq}$6.5 cm) were differentiated into an E2 group and an E2-free group and then they were reared for about four months at three water temperature levels of $20^{\circ}C$, $24^{\circ}C$, and $28^{\circ}C$. The results showed that the young eels survived normally at the rearing water temperature of ${\geq}24^{\circ}C$, and grew to a mean size of 20 cm (total length). In the E2-free group, temperature was not found to increase the sex ratio (feminizing rates); however, the sex ratio of the E2-administrated group was found to be a little higher at a high temperature ($28^{\circ}C$). The growth of the E2 group was lower than the growth of the E2-free group at $24^{\circ}C$ and the E2 concentration levels in the plasma at $24^{\circ}C$ were found to be significant after the end of the E2 administration period (178 days). Therefore, we thought that long-term administration of E2 must be considered to be the reason for growth decline in spite of the prominent sex ratio effect. Our results indicate that temperature was not related to an increase in the feminizing rate (sex ratio) in the Japanese eel, Anguilla japonica, and other environmental factors (rearing density, salinity, etc.) that have the possibility of inducing ovarian differentiation must be investigated.

• Development and Reproduction
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• v.23 no.4
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• pp.333-344
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• 2019

In vertebrate reproductive system, estrogen receptor (ER) plays a pivotal role in mediation of estrogenic signaling pathways. In the present study, we report the cDNA cloning, expression analysis, and transcriptional activity of ERβ1 subtype from medaka Oryzias dancena. The deduced O. dancena ERβ1 (odERβ1; 519 amino acids) contained six characteristic A/B to E/F domains with very short activation function 2 region (called AF2). A phylogenetic analysis indicated that odERβ1 was highly conserved among teleost ERβ1 subgroup. A conventional RT-PCR revealed that the odERβ1 transcripts were widely distributed in the multiple tissues, the ovary, brain, gill, intestine, kidney, and muscle. Further, the relatively higher odERβ1 expressions in the ovary and brain were clearly reproduced in RT-qPCR assay. When HA-fused odERβ1 expression vector was transfected into HEK293 cells, an immunoreactivity for odERβ1 was mainly detected in the nucleus part. Finally, an estrogen responsive element driven luciferase reporter assays demonstrated that the transcriptional activity of odERβ1 significantly increased by estradiol-17β (E2) in a dose dependent manner (p<0.05). However, fold-activation of odERβ1 in the presence of E2 was markedly weak, when it compared with those of O. latipes ERβ1. Taken together, these data suggest that odERβ1 represents a functional variant of teleost ERβ subtype and provides a basic tool allowing future studies examining the function of F domain of ERβ1 subtype and expanding our knowledge of ERβ evolution.

• Korean Journal of Veterinary Research
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• v.36 no.1
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• pp.93-99
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• 1996

This study was designed to investigate the effect of estrogen(Est) on the progestcrone(Prog) target cells by autoradiography. The spayed 16 mice(ICR, approximately 18~25g) were randomly alloted into 3 groups. $^3H$-Prog-treated group were injected with $40{\mu}Ci$ of $^3H$-Prog/mouse/day for 1 day, Est + $^3H$-Prog-treated group with $20{\mu}Ci$ of $17{\beta}$-Est/mouse/day for 3 days and then with $40{\mu}Ci$ of $^3H$-Prog/mouse at 4th day, and Est+$^3H$-thymidine(TdR)-treated group with $20{\mu}g$ of $17{\beta}$-Est/mouse/day for 3 days and then $80{\mu}Ci$ of $^3H$-TdR/mouse at 4th days. 1. Mice uteri of both Est+$^3H$-Prog-treated group and Est+$^3H$-TdR-treated group were hypemophied in gross finding and the endometrium and myometrium were thickened in microscopic findings. These findings were confirmed that Est enlarged the uteri of mice. 2. Cryo-preparations of mice organs were processed for autoradiography using Kodak NTB-2 emulsion following Kodak D-19 developer and hematoxylin counterstain. In each group, the number values of silver grain distribution appeared to be higher in the $^3H$-Prog-treated group than in the Est+$^3H$-Prog-treated group. It was considered that Est and Prog inhibit each other in action. 3. In both $^3H$-Prog-treated group and Est+$^3H$-Prog-treated group, the uteri have highest distribution rates of silver grains than in other organs, and the cerebral neurons, hepatocytes, bronchiolar epithelial cells and splenic reticular cells also contained some silver grains. 4. The orders of the cell types with more number of silver grains in the uteri were stromal cells, glandular epithelial cells, luminal surface cells and muscular cells and also were as above orders in distribution of proliferating cell type by $^3H$-TdR.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.2
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• pp.113-123
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• 2022

Diarylpropionitrile (DPN), a selective agonist for estrogen receptor β (ERβ), has been reported to regulate various hormonal responses through activation of ERβ in tissues including the mammary gland and brain. However, the effect of DPN on melanogenesis independent of ERβ has not been studied. The aim of this study is to examine the possibility of anti-melanogenic effect of DPN and its underlying mechanism. Melanin contents and cellular tyrosinase activity assay indicated that DPN inhibited melanin biosynthesis in alpha-melanocyte stimulating hormone-stimulated B16F10 melanoma cell line. However, DPN had no direct influence on in vitro tyrosinase catalytic activity. On the other hand, 17β-estradiol had no effect on inhibition of melanogenesis, suggesting that the DPN-mediated suppression of melanin production was not related with estrogen signaling pathway. Immunoblotting analysis showed that DPN down-regulated the expression of microphthalmia-associated transcription factor (MITF), a central transcription factor of melanogenesis and its down-stream genes including tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2. Also, DPN attenuated the phosphorylation of protein kinase A (PKA) and cAMP-response element-binding protein (CREB). Additionally, DPN suppressed the melanin synthesis in UVB-irradiated HaCaT conditioned media culture system suggesting that DPN has potential as an anti-melanogenic activity in physiological conditions. Collectively, our data show that DPN inhibits melanogenesis via downregulation of PKA/CREB/MITF signaling pathway.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.4-4
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• 2023

다국적 제약회사에서 생산되는 제약화합물은 인간의 질병치료 뿐만 아니라 가축의 생산성을 향상시키는 물질으로도 활발하게 생산되는데 육우의 성장을 촉진하는 기능성 화합물들도 많이 있다. 북미에서 생산되는 소고기의 약 80%가 1번 이상의 성장촉진기술들이 활용되고 있다고 보고된다. 고기소로 생산되는 비육우에 적용되는 방법은 크게 피하 이식에 의해 혈류를 타고 성장을 촉진하는 17β-Estradiol과 합성 남성호르몬제인 Trenbolone acetate가 주로 활용되고, 비육후기 사료에 섞어서 급여하는 사료첨가제 형태인 β2-adrenergic agonist 같은 형태로 적용하게 된다. 근육을 성장하는 기술의 작용기전은 많은 선행연구에 의해 밝혀져 있는 반면 천연 알칼로이드 성분들의 기전은 밝혀진 것이 많지 않다. 한방제재들에서 많이 발견되는 알칼로이드 성분들은 생리활성 기능들을 가진 것으로 알려져 있지만, 생산, 수거, 가공, 추출 등의 공정에서 많은 비용이 발생하므로 비육우의 사료화 가능성은 아주 희박하다. 따라서 비용을 최소화 할 수 있는 후보재료를 검색중에 감자부산물을 확보하였고 기능성 물질의 추출과 사료첨가제 화 하여 비육우에 급여시험을 실시하였다. α-solanine과 α-chaconine은 감자의 잎, 과일 및 괴경에서 발견되는 글리코알칼로이드 화합물로, 쥐, 토끼, 닭과 같은 다양한 동물 모델에서 중독성을 가진 물질로 보고 되고 있다. 최근 연구에서는 비육우의 성능을 유도하는 데 사용되는 것으로 나타났다. 한우 송아지 3마리의 사태(Semimembranosus)와 등심(Longissimus Dorci)근육에서 추출된 근육위성 세포(BSC)에 다양한 수준의 α-solanine(control, 0.001, 0.01, 0.1, 1, 10μM)으로 처리해본 결과 근육관련 지표인 MHC2X과 β2-AR의 발현이 높게 나타난 것을 확인했다. 사료급여실험에서는 대조군에 비해 급여군의 등심단면적과 도체중이 향상되는 것을 확인하였다. 결과적으로 감자유래 농산부산물은 한우 비육우의 근육의 성장을 증가시키고, 그 작용기전은 β2-수용체에 작용하여 단백질 합성을 촉진시켜 근육을 축적시키는 것으로 확인하였다. 농산부산물을 이용한 기능성 사료개발은 최근 이슈가 되고 있는 축산분야 탄소저감을 개선할 수 있는 기술로 축산의 업사이클링 기술로 활용 가능하다.

• Korean Journal of Agricultural Science
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• v.35 no.2
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• pp.167-176
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• 2008

Supplementation of serum and estrogen in in vitro maturation(IVM) medium was shown to improve embryo development and quality in several species. This study investigates the effect of ovarian estrus stage on canine oocyte quality and supplementation of medium with canine serum or estrogen on IVM of canine oocytes. As results, in experimental 1, IVM oocytes collected from follicular stage ovaries to MII stages($10.2{\pm}1.5%$) was higher (p<0.05) with 10% canine estrus stage serum than control($1.3{\pm}1.6%$), anoestrus stage serum($4.0{\pm}1.6%$), luteal stage serum($2.7{\pm}1.7%$) and 10% FBS($1.3{\pm}1.6$). In experimental 2, 10% canine estrus stage serum supplementation has highest maturation rate to MII stages($10.0{\pm}1.8%$) and there were significant differences(P<0.05) with another treatment in follicular stages group. In order to investigate the synergic effect of estrous serum and estrogen supplementation, different estrous stage groups of oocytes were cultured with 2 ug/ml estrogen plus various concentrations of different reproductive stage serum and FBS(experimental 3). As results, the rate of maturation to metaphase II(MII) stage was significantly higher(p<0.05) in oocytes from the follicular stage supplemented with estrogen and 10% canine estrus stage serum(11.5%) compared to the other groups(6.0 - 8.8%). The present study was demonstrated that canine serum and the estrus cycle of the bitch affect the meiotic competence of oocytes. Hormonal influences within the follicle may be one of the factors responsible for the greater proportion of maturation of oocyte to MII from bitches at the follicular phase.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.209-220
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• 2017

The estrogen-mediated effect of mesenchymal stem cells (MSCs) is a highly critical factor for the clinical application of MSCs. However, the present study is conducted on MSCs derived from adult donors, which have different physiological status with steroid hormonal changes. Therefore, we explores the important role of $17{\beta}$-estradiol (E2) in MSCs derived from female and male newborn piglets (NF- and NM-pBMSCs), which are non-sexually matured donors with steroid hormones. The results revealed that in vitro treatment of MSCs with E2 improved cell proliferation, but the rates varied according to the gender of the newborn donors. Following in vitro treatment of newborn MSCs with E2, mRNA levels of Oct3/4 and Sox2 increased in both genders of MSCs and they may be correlated with both estrogen receptor ${\alpha}$ ($ER{\alpha}$) and $ER{\beta}$ in NF-pBMSCs, but NM-pBMSCs were only correlated with $ER{\alpha}$. Moreover, E2-treated NF-pBMSCs decreased in ${\beta}$-galactosidase activity but no influence on NM-pBMSCs. In E2-mediated differentiation capacity, E2 induced an increase in the osteogenic and chondrogenic abilities of both pBMSCs, but adipogenic ability may increased only in NF-pBMSCs. These results demonstrate that E2 could affect both genders of newborn donor-derived MSCs, but the regulatory role of E2 varies depending on gender-dependent characteristics even though the original newborn donors had not been affected by functional steroid hormones.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.3
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• pp.194-206
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• 2020

Objective: The aim of this study was to investigate microRNAs (miRNAs) related to follicle-stimulating hormone (FSH) responsiveness using miRNA microarrays and to identify their target genes to determine the molecular regulatory pathways involved in FSH signaling in KGN cells. Methods: To change the cellular responsiveness to FSH, KGN cells were treated with FSH receptor (FSHR)-specific small interfering RNA (siRNA) followed by FSH. miRNA expression profiles were determined through miRNA microarray analysis. Potential target genes of selected miRNAs were predicted using bioinformatics tools, and their regulatory function was confirmed in KGN cells. Results: We found that six miRNAs (miR-1261, miR-130a-3p, miR-329-3p, miR-185-5p, miR-144-5p and miR-4463) were differentially expressed after FSHR siRNA treatment in KGN cells. Through a bioinformatics analysis, we showed that these miRNAs were predicted to regulate a large number of genes, which we narrowed down to cytochrome P450 family 19 subfamily A member 1 (CYP19A1) and estrogen receptor alpha (ESR1) as the main targets for miR-4463. Functional analysis revealed that miR-4463 is a regulatory factor for aromatase expression and function in KGN cells. Conclusion: In this study, we identified differentially expressed miRNAs related to FSH responsiveness. In particular, upregulation of miR-4463 expression by FSHR deficiency in human granulosa cells impaired 17β-estradiol synthesis by targeting CYP19A1 and ESR1. Therefore, our data might provide novel candidates for molecular biomarkers for use in research into poor responders.