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Identification of Egr1 Direct Target Genes in the Uterus by In Silico Analyses with Expression Profiles from mRNA Microarray Data

  • Seo, Bong-Jong;Son, Ji Won;Kim, Hye-Ryun;Hong, Seok-Ho;Song, Haengseok
    • Development and Reproduction
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    • v.18 no.1
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    • pp.1-11
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    • 2014
  • Early growth response 1 (Egr1) is a zinc-finger transcription factor to direct second-wave gene expression leading to cell growth, differentiation and/or apoptosis. While it is well-known that Egr1 controls transcription of an array of targets in various cell types, downstream target gene(s) whose transcription is regulated by Egr1 in the uterus has not been identified yet. Thus, we have tried to identify a list of potential target genes of Egr1 in the uterus by performing multi-step in silico promoter analyses. Analyses of mRNA microarray data provided a cohort of genes (102 genes) which were differentially expressed (DEGs) in the uterus between Egr1(+/+) and Egr1(-/-) mice. In mice, the frequency of putative EGR1 binding sites (EBS) in the promoter of DEGs is significantly higher than that of randomly selected non-DEGs, although it is not correlated with expression levels of DEGs. Furthermore, EBS are considerably enriched within -500 bp of DEG's promoters. Comparative analyses for EBS of DEGs with the promoters of other species provided power to distinguish DEGs with higher probability as EGR1 direct target genes. Eleven EBS in the promoters of 9 genes among analyzed DEGs are conserved between various species including human. In conclusion, this study provides evidence that analyses of mRNA expression profiles followed by two-step in silico analyses could provide a list of putative Egr1 direct target genes in the uterus where any known direct target genes are yet reported for further functional studies.

Expression Pattern of Early Growth Response Gene 1 during Olive Flounder (Paralichthys olivaceus) Embryonic Development

  • Yang, Hyun;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Kim, Kyung-Kil
    • Development and Reproduction
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    • v.18 no.4
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    • pp.233-240
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    • 2014
  • The early growth response protein 1 (Egr-1) is a widely reported zinc finger protein and a well known transcription factor encoded by the Egr-1 gene, which plays key roles in many aspects of vertebrate embryogenesis and in adult vertebrates. The Egr-1 expression is important in the formation of the gill vascular system in flounders, which develops during the post-hatching phase and is essential for survival during the juvenile period. However, the complete details of Egr-1 expression during embryo development in olive flounder are not available. We assessed the expression patterns of Egr-1 during the early development of olive flounders by using reverse transcription polymerase chain reaction (RT-PCR) analysis. Microscopic observations showed that gill filament formation corresponded with the Egr-1 expression. Thus, we showed that Egr-1 plays a vital role in angiogenesis in the gill filaments during embryogenesis. Further, Egr-1 expression was found to be strong at 5 days after hatching (DAH), in the development of the gill vascular system, and this strong expression level was maintained throughout all the development stages. Our findings have important implications with respect to the biological role of Egr-1 and evolution of the first respiratory blood vessels in the gills of olive flounder. Further studies are required to elucidate the Egr-1-mediated stress response and to decipher the functional role of Egr-1 in developmental stages.

Ameliorative Effects of Soybean Leaf Extract on Dexamethasone-Induced Muscle Atrophy in C2C12 Myotubes and a C57BL/6 Mouse Model (콩잎 추출물의 근위축 개선 효과)

  • Hye Young Choi;Young-Sool Hah;Yeong Ho Ji;Jun Young Ha;Hwan Hee Bae;Dong Yeol Lee;Won Min Jeong;Dong Kyu Jeong;Jun-Il Yoo;Sang Gon Kim
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1036-1045
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    • 2023
  • Sarcopenia, a condition characterized by the insidious loss of skeletal muscle mass and strength, represents a significant and growing healthcare challenge, impacting the mobility and quality of life of aging populations worldwide. This study investigated the therapeutic potential of soybean leaf extract (SL) for dexamethasone (Dexa)-induced muscle atrophy in vitro and in an in vivo model. In vitro experiments showed that SL significantly alleviated Dexa-induced atrophy in C2C12 myotube cells, as evidenced by preserved myotube morphology, density, and size. Moreover, SL treatment significantly reduced the mRNA and protein levels of muscle RING-finger protein-1 (MuRF1) and muscle atrophy F-box (MAFbx), key factors regulating muscle atrophy. In a Dexa-induced atrophy mouse model, SL administration significantly inhibited Dexa-induced weight loss and muscle wasting, preserving the mass of the gastrocnemius and tibialis anterior muscles. Furthermore, mice treated with SL exhibited significant improvements in muscle function compared to their counterparts suffering from Dexa-induced muscle atrophy, as evidenced by a notable increase in grip strength and extended endurance on treadmill tests. Moreover, SL suppressed the expression of muscle atrophy-related proteins in skeletal muscle, highlighting its protective role against Dexa-induced muscle atrophy. These results suggest that SL has potential as a natural treatment for muscle-wasting conditions, such as sarcopenia.

Event Related Potential and Neuropsychological Tests (사건 관계 전위와 신경심리 검사)

  • Lee, Sung-Hoon;Koo, Ae-Sook;Woo, Jong-Ho;Kim, Byung-Gun
    • Sleep Medicine and Psychophysiology
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    • v.1 no.1
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    • pp.76-86
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    • 1994
  • Objectives: This research was performed in order to observe the neuropsychological implication and functional anatomical source of event related potentials(ERP) by studying of correlations between event related potentials and neuropsychological tests. Methods: The latency and voltage of P100 of visual evoked potential (VEP), and N120 and P300 of event related potentials were studied in 56 patients and their correlations with neuropsychological tests were computed. Results: The tests showing significant correlation with latency P100 were visual continous performance test(VCPT) and contingent continous performance test(CCPT) without any significant correlation with voltage of P100. In latency of N120 category test and verbal IQ of KWIS showed significant correlation, and in voltage of N120, finger tapping test, VCPT, CCPT and digit symbol test displayed significant correlations. The latency of P300 had significant correlation with trail making A test and Stroop test. In the voltage of P300 significant correlations were shown with trail making B test, digit symbol test and Wechsler memory scale, finger tapping test, stroop test, VCPT and CCPT. Conclusion : N120 may be considered to reflect the function of medial frontal lobe and P300 may be considered to be developed from the subcortical connection of medial temporal lobe, hippocampus, thalamus, basal ganglia and medial frontal lobe.

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Identification of Fruit-specific cDNAs in a Ripened Inodorus Melon Using Differential Screening and the Characterization of on Abscisic Acid Responsive Gene Homologue

  • Hong, Se-Ho;Kim, In-Jung;Chung, Won-Il
    • Journal of Plant Biotechnology
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    • v.4 no.1
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    • pp.7-15
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    • 2002
  • Eight cDNAs corresponding to fruit-specific genes were isolated from ripened melon through differential screening. Sequence comparison indicated that six of these cDNAs encoded proteins were previously characterized into aminocyclopropane-1-carboxylate (ACC) oxidase, abscisic acid, stress and ripening inducible (ASR) gene, RINC-H2 zinc finger protein, pyruvate decarboxylase, or polyubiquitin. RFS2 and RFS5 were the same clone encoding polyubiquitin. The other cDNAs showed no significant homology with known protein sequences. The ASR homologue (Asr1) gene was further characterized on the cDNA and genomic structure. The deduced amino acid sequence had similar characteristics to other plant ASR. The Asr1 genomic DNA consisted of 2 exons and 1 intron, which is similar to the structure of other plants ASR genes. The promoter region of the Asr1 gene contained several putative functional cis-elements such as an abscisic acid responsive element (ABRE), an ethylene responsive element (ERE), a C-box or DPBf-1 and 2, Myb binding sites, a low temperature responsive element (LTRE) and a metal responsive element (MRE). The findings imply that these elements may play important roles in the response to plant hormones and environmental stresses in the process of fruit development. The results of this study suggest that the expressions of fruit specific and ripening-related cDNAs are closely associated with the stress response.

Power-Dependent Characteristics of $n^+$-p and $p^+$-n GaAs Solar Cells

  • Kim, Seong-Jun;Kim, Yeong-Ho;No, Sam-Gyu;Kim, Jun-O;Lee, Sang-Jun;Kim, Jong-Su;Lee, Gyu-Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.08a
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    • pp.236-236
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    • 2010
  • 단일접합 $n^+-p/p^+$ (p-emitter) 및 $p^+-n/n^+$ (n-emitter) GaAs 태양전지 (Solar Cell)를 각각 제작하여, 그 소자특성을 비교 분석하였다. AM 1.5 (1 sun, $100\;mW/cm^2$) 표준광을 조사할 경우, p-emitter/n-emitter 소자의 개방회로전압 (Voc), 단락회로전류 (Jsc), 충전율 (FF), 효율 (Eff)은 각각 0.910/0.917 V, $15.9/16.1\;mA/cm^2$, 78.7/78.9, 11.4/12.1%로서, n-emitter 소자가 다소 크지만 거의 비슷한 값을 가지고 있었다. 태양전지의 집광 특성을 분석하기 위하여 조사광의 출력에 따른 태양전지의 소자 특성을 측정하였다. 조사광 강도가 높아짐에 따라 p-emitter 소자의 특성은 점진적으로 증가하는 반면, n-emitter는 1.3 sun에서 약 1.4 배의 최대 효율 (17%)을 나타내고 조사광이 더 증가함에 따라 급격히 감소하는 특성을 보여 주었다. (그림 참고) 본 연구에서 사용한 2종류 소자의 층구조는 서로 반대되는 대칭구조로서, 모두 가까이에 위치하고 있는 표면전극 (surface finger) 방향으로 소수전하 (minority carrier)가 이동하고 다수전하 (majority carrier)는 기판 (두께 $350\;{\mu}m$)을 통한 먼 거리의 후면전극 (back electrode)으로 표류 (drift)되도록 설계되어 있다. 이때, n-emitter에서는 이동도 (mobility)와 확산길이 (diffusion length)가 높은 전자가 후면전극으로 이동하기 때문에 적정밀도의 전자-정공 쌍 (EHP)이 여기될 경우에는 Jsc와 Eff가 극대화되지만, 조사광 강도 또는 EHP가 더 높아질 경우에는 직렬저항의 증가와 함께 전류-전압 (I-V)의 이상인자 (ideality factor)가 커짐으로서 FF와 효율이 급격히 감소한 결과로 분석된다. 현재 전산모사를 통한 자세한 분석을 진행하고 있으며, 본 결과는 효율 극대화를 위한 최적 층구조 및 도핑 밀도 설계에 활용할 수 있을 것으로 판단된다.

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Methylation Status of H19 Gene in Embryos Produced by Nuclear Transfer of Spermatogonial Stem Cells in Pig

  • Lee, Hyun-Seung;Lee, Sung-Ho;Gupta, Mukesh Kumar;Uhm, Sang-Jun;Lee, Hoon-Taek
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.67-75
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    • 2011
  • The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFR ${\alpha}$-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs ($11.2{\pm}0.8%$) and SSCs ($13.3{\pm}1.1%$). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.

Role of IL-15 in Sepsis-Induced Skeletal Muscle Atrophy and Proteolysis

  • Kim, Ho Cheol;Cho, Hee-Young;Hah, Young-Sool
    • Tuberculosis and Respiratory Diseases
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    • v.73 no.6
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    • pp.312-319
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    • 2012
  • Background: Muscle wasting in sepsis is associated with increased proteolysis. Interleukin-15 (IL-15) has been characterized as an anabolic factor for skeletal muscles. Our study aims to investigate the role of IL-15 in sepsis-induced muscle atrophy and proteolysis. Methods: Mice were rendered septic either by cecal ligation and puncture or by intraperitoneal injection of lipopolysaccharide (LPS, 10 mg/kg i.p.). Expression of IL-15 mRNA and protein was determined by reverse transcriptase polymerase chain reaction and Western blot analysis in the control and septic limb muscles. C2C12 skeletal muscle cells were stimulated in vitro with either LPS or dexamethasone in the presence and absence of IL-15 and sampled at different time intervals (24, 48, or 72 hours). IL-15 ($10{\mu}g/kg$) was intraperitoneally administered 6 hours before sepsis induction and limb muscles were sampled after 24 hours of sepsis. Cathepsin L activity was determined to measure muscle proteolysis. Atrogin-1 and muscle-specific ring finger protein 1 (MuRF1) expressions in limb muscle protein lysates was analyzed. Results: IL-15 mRNA expression was significantly lower in the limb muscles of septic mice compared to that of controls. Cathepsin L activity in C2C12 cells was significantly lower in presence of IL-15, when compared to that observed with individual treatments of LPS or dexamethasone or tumor necrosis factor ${\alpha}$. Further, the limb muscles of mice pre-treated with IL-15 prior to sepsis induction showed a lower expression of atrogin-1 and MuRF1 than those not pre-treated. Conclusion: IL-15 may play a role in protection against sepsis-induced muscle wasting; thereby, serving as a potential therapeutic target for sepsis-induced skeletal muscle wasting and proteolysis.

Studies on the Fabrication and Characteristics of PHEMT for mm-wave (mm-wave용 전력 PHEMT제작 및 특성 연구)

  • Lee, Seong-Dae;Chae, Yeon-Sik;Yun, Gwan-Gi;Lee, Eung-Ho;Lee, Jin-Gu
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.38 no.6
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    • pp.383-389
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    • 2001
  • We report on the design, fabrication, and characterization of 0.35${\mu}{\textrm}{m}$-gate AIGaAs/InGaAs PHEMTs for millimeter-wane applications. The epi-wafer structures were designed using ATLAS for optimum DC and AC characteristics, 0.351m-gate AIGaAs/rnGaAs PHEMTs having different gate widths and number of fingers were fabricated using electron beam lithography Dependence of RF characteristics of PHEMT on gate finger with and number of gate fingers have been investigated. PHEMT haying two 0.35$\times$60${\mu}{\textrm}{m}$$^2$ gate fingers showed the knee voltage, pinch-off voltage, drain saturation current density, and maximum transconductance of 1.2V, -1.5V, 275㎃/mm, and 260.17㎳/mm, respectively. The PHEMT showed fT(equation omitted)(current gain cut-off frequency) of 45㎓ and fmax(maximum oscillation frequency) of 100㎓. S$_{21}$ and MAG of the PHEMT were 3.6dB and 11.15dB, respectively, at 35㎓

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Aryl Sulfonamides Induce Degradation of Aryl Hydrocarbon Receptor Nuclear Translocator through CRL4DCAF15 E3 Ligase

  • Kim, Sung Ah;Jo, Seung-Hyun;Cho, Jin Hwa;Yu, Min Yeong;Shin, Ho-Chul;Kim, Jung-Ae;Park, Sung Goo;Park, Byoung Chul;Kim, Sunhong;Kim, Jeong-Hoon
    • Molecules and Cells
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    • v.43 no.11
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    • pp.935-944
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    • 2020
  • Aryl hydrocarbon receptor nuclear translocator (ARNT) plays an essential role in maintaining cellular homeostasis in response to environmental stress. Under conditions of hypoxia or xenobiotic exposure, ARNT regulates the subset of genes involved in adaptive responses, by forming heterodimers with hypoxia-inducible transcription factors (HIF1α and HIF2α) or aryl hydrocarbon receptor (AhR). Here, we have shown that ARNT interacts with DDB1 and CUL4-associated factor 15 (DCAF15), and the aryl sulfonamides, indisulam and E7820, induce its proteasomal degradation through Cullin-RING finger ligase 4 containing DCAF15 (CRL4DCAF15) E3 ligase. Moreover, the two known neo-substrates of aryl sulfonamide, RNA-binding motif protein 39 (RBM39) and RNA-binding motif protein 23 (RBM23), are not required for ARNT degradation. In line with this finding, aryl sulfonamides inhibited the transcriptional activities of HIFs and AhR associated with ARNT. Our results collectively support novel regulatory roles of aryl sulfonamides in both hypoxic and xenobiotic responses.