• Title/Summary/Keyword: mRNA 발현

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생쥐 초기 배아에서 Aquaporin 8과 9의 발현에 관한 연구

  • 신현상;계명찬;강수만;이성은;이지원;강한승;김문규
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.68-68
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    • 2003
  • Aquaporin은 막관통 통로 단백질(transmembrane channel protein)로서, 삼투압의 농도구배에 따라 세포막을 가로질러 물분자를 이동시키는 기능을 하고 있다. 포유류 초기배아에서 포배강 형성은 영양외배엽세포에서 $Na^+ / K^+$ATPase에 의한 이온 농도 구배가 형성되면 auqaporin에 의해 물이 포배강으로 유입되면서 이루어진다. 본 연구에서는 생쥐 초기배아에서 반정량적인 역전사 중합효소 연쇄반응 방법(semi-quantitative RT-PCR)과 실시간 역전사 중합효소 연쇄반응 방법(real-time RT-PCR)을 통하여 AQP8과 9의 mRNA발현을 조사하고 다중 면역형광현미경 방법(confocal immunofluorescence microscopy)을 통해 단백질 발현양상을 분석하였다. AQP8 mRNA는 상실기까지 발현되지 않다가 포배기에 이르러 발현되었고 AQP9 mRNA는 수정란에서부터 발현되어 포배기에는 유의할 정도로 증가하였다. 따라서 AQP8 mRNA는 배아유전자가 활성화되어 나타나는 것이고 AQP9 mRNA는 모계유전자 기원임을 알 수 있었다. AQP8 단백질은 상실배 단계까지 발현되지 않다가 포배시기에 영양외배엽세포사이의 접합면에 발현되었고 AQP9 단백질은 상실배 시기에 할구 사이의 인접 부위에서 강하게 발현되었다가 포배시기에는 세포간의 접합면에 약하게 발현하는 경향을 나타내었다. 실시간 역전사 중합효소 연쇄반응 방법으로 조사한 결과 포배에서 물과 글리세롤을 통과시키는 AQP9는 mRNA의 발현양이 AQP8보다 약 4배 정도 많았다. 또한 포배기에 이르러서야 물만을 통과시키는 AQP8의 발현이 나타나는 것을 보아 포배강 형성시 외부에서 영양외배엽을 통해 포배강으로 유입되는 물의 이동(trans- trophectodermal water movements)에 AQP9보다 AQP8이 더 중요하게 관여할 것으로 사료된다., K, Pb, Cd, Cr, Co, Cu, Ni)을 측정하였다. 실험 조건1의 결과로서 각 국의 유아용 일회용 기저귀의 중금속 함량은 거의 유사한 경향을 나타내었으며 Cr, Zn, Pb, Ni, Mn, Mg, Li, K는 detection limit(2 ppm) 이하였고, Cd, Fe, Co, Cu, Ca, Al, Sr는 검출되었지만 기준치 이하였다. 실험 조건2의 결과로서 측정 항목(Cr, Sb, Cd, Pb, Ni, Co, Cu)중 Cr, Cd, Ni, Cu는 detection limit(0.1 ppm) 이하였고, Sb, Pb, Co는 검출되었지만 기준치 이하였다.았다. 4%의 경우에는 8$0^{\circ}C$이하로 온도를 낮추는 것이 좋은 상태를 나타내었다. 이와 같은 결과는 일반적으로 화학적 레팅을 4%, 7%에서한 선행결과와 상당히 다른 결과이다.염 농도가 증가할수록 감소 현상을 보였다.X>, 75BG30은 8.6$\mu\textrm{m}$, 75BG40은 7.02$\mu\textrm{m}$로 나타났다. 따라서 경화제 양에 관계없이 10$\mu\textrm{m}$ 이하로 나타나, 경화제 10$m\ell$만으로 미세한 크기를 얻을 수 있음을 알 수 있다. 젤리 강도 변화에 따른 차이는 300BF는 78.09$\mu\textrm{m}$ 300BG는 56.32$\mu\textrm{m}$로, 75BF나 75BG에 비하여 현저히 증가하여, 젤라틴의 젤리 강도는 캡슐 제조 조건의 주요한 변수임을 알 수 있다.추출물 투여시 혈당강하 및 혈중콜레스테롤 강하가 나타났으며, 상엽복합추출물 투여와 운동을 병행시 이러한 감소 효과가 더 뚜렷하게 나타났다.교육의 적임자로 보는 시각이 비교적 높았고 약 1/2정도는 영양교육에 참여하겠다는 의지를 가지고 있을 뿐만 아니라 실제로 영양지도를

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Role of Lysyl Oxidase Family during Odontoblastic Differentiation of Human Dental Pulp Cells Induced with Odontogenic Supplement (인간치수세포에서 상아모세포의 분화과정 동안 Lysyl Oxidase Family의 역할)

  • Lee, Hwa-Jeong;Han, Soo-Yeon
    • Journal of dental hygiene science
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    • v.13 no.3
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    • pp.296-303
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    • 2013
  • Lysyl oxidase (LOX), extracellular matrix enzyme, is catalyzing lysine-derived crosslinks in collagen and elastin. Recently, several LOX-like proteins (LOXL, LOXL2, LOXL3 and LOXL4) have been identified in human but their specific functions are still largely unknown. The purpose of this study was to evaluate the function of the LOX family genes during odontoblastic differentiation of human dental pulp (HDP) cells induced with odontogenic supplement (OS). The messenger RNA (mRNA) expression of LOX family genes and differentiation markers was assessed by reverse transcriptase polymerase chain reaction analysis (RT-PCR). The formation of mineralization nodules was evaluated by alrizarin red S staining. Amine oxidase activity of HDP cells was measured by peroxidase-coupled fluormetric assay. The expressions of differentiation markers, such as alkaline phosphatase (ALP), osteopontin (OPN), osteocalcin (OCN), dentin matrix protein1 (DMP1), dentin sialophosphoprotein (DSPP) in HDP cells were increased after treatment with OS media. The LOX and LOXL mRNA expression were gradually increased in OS media, whereas LOX enzyme activities were markedly detected on day 7. The mRNA expression and LOX enzyme activity of collagen type I was very similar to the pattern of LOX gene. In this study, the expression of LOX and its isoforms, and activity of LOX were highly regulated during odontoblastic differentiation. Thus, these results suggest that LOX plays a key role in odontoblastic differentiation of HDP cells.

Expression of Growth Differentiation Factor-9 in the Mouse Ovaries at Different Developmental Stages (생쥐 난소의 발생단계에 따른 Growth Differentiation Factor-9의 유전자 발현)

  • 윤세진;이경아;고정재;차광열
    • Development and Reproduction
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    • v.3 no.1
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    • pp.95-100
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    • 1999
  • Growth differentiation factor-9 (GDF-9) is a member of the transforming growth factor $\beta$ (TGF-$\beta$) superfamily. It has been known that GDF-9 is a growth factor having a crucial role in normal folliculogenesis and its expression is oocyte-specific. The present study was aimed to elucidate the expression of GDF-9 mRNA in the mouse primordial follicles as well as in the other developmental stages. The semiquantitative analysis of GDF-9 mRNA expression was conducted. Total RNA was extracted from the ICR mice ovaries at gestational day 19, postnatal day 1, day 10, day 21, and day 28, and RT-PCR was performed to measure GDF-9 and $\beta$-actin mRNA levels. Level of GDF-9 mRNA were normalized against the level of $\beta$-actin mRNA, and compared among different stages. GDF-9 mRNA was detected in all samples including the fetal ovaries that mainly consists of primordial follicles. The highest level of mRNA was observed in ovaries obtained at day 10 that mainly consists of growing follicles. The present result suggests that GDF-9 may play an important role in the early stage of folliculogenesis.

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Expression of mRNAs characteristic of cartilage and bone in the developing mandibular condyle of mice (발육중인 생쥐 하악 과두에서 연골 및 골의 특이 유전자 발현)

  • Ji, Kuk-Soep;Yoon, Young-Jooh;Park, Joo-Cheol;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.34 no.2 s.103
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    • pp.143-152
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    • 2004
  • It has not been elucidated whether the initiation of condylar development of the mandible is related with the periosteum of the mandible, or if it derives from a separate programmed blastema not related with the mandible. Also, although the mandibular condylar cartilage is known to promote growth, few studies have dealt with molecular-biologic mechanisms such as the expression of specific genes according to the differentiation of the mandibular condyle. To elucidate the unique cellular characteristics, development, and differentiation process of the mandibular condyle, an examination of expressions of genes characteristic of cartilage and bone were carried out using RT-PCR and mRNA in situ hybridization. 1. Type? collagen mRNA was detected with type II collagen mRNA in the differentiation and growth process of the cartilage of the mandibular condyle. TypeII collagen mRNA was demonstrated in the whole resting md upper part of the poliferative zone, whereas type II collagen mRNA was observed in the resting, proliferative and upper hypertrophic cartilage zone of the mandibular condyle. 2. The condylar cartilage rapidly increased in size due to the accumulation of hypertrophic chondrocytes as characterized by the expression of type II collagen mRNA during postnatal development. 3. BMP-4 mRNA was present in the anlage of the future condylar process and also in the ossifying mandibular body. 4. IHH mRNA was limited exclusively to the lower part of the proliferative zone and the upper part of the hypertrophic cartilage zone during condylar development. These findings were different from those in the growth-plate cartilage of the long bone, indicating a characteristic feature of the differentiation of the chondrocytes in the condylar cartilage present in prenatal and postnatal development. Furthermore, it was also suggested that chondroblasts of condylar cartilage rapidly differentiate into hypertrophic chondrocytes with increased functional Load force such as muscle activity and mastication.

Eotaxin mRNA Expression in Bronchial Mucosa of Patients with Asthma (천식 환자의 기관지 조직에서 Eotaxin mRNA 발현에 관한 연구)

  • In, Kwang-Ho;Cho, Jae-Yun;Kang, Sae-Yong;Lee, Sang-Youb;Shim, Jae-Jeong;Kang, Kyung-Ho;Yoo, Se-Hwa;Na, Young-Soon;Kim, Han-Gyum
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.697-704
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    • 1998
  • Background: Asthma is a chronic inflammatory disease of the airways characterized by a marked infiltration of eosinophils in the bronchial mucosa. Asthmatic bronchial mucosa produces many factors described as being chemotactic for inflammatory cells. IL-5, RANTES, and MCP-1 alpha are the chemotactic factors for eosinophils, but their roles are controversial. Recently eotaxin that is a potent eosinophil chemoattractant cytokine was detected in a guinea-pig model of allergic airway inflammation, and human eotaxin was cloned. Eotaxin is a specific chemoattractant for eosinophils, but its role in asthma is not confirmed. We examined the in vivo expression of eotaxin in bronchi of asthmatic patients. Methods : 11 asthmatics and 2 normal controls were enrolled. All subjects were underwent bronchoscopy with bronchial biopsies in 2nd or 3rd carina. RNA extraction from biopsy samples was done by acid-guanidium method. Semi-quantitaive RT-PCR was done for evaluation of eotaxin mRNA expression The extent of eosinophil infiltration was evaluated by counting the eosinophils in submucosa in HPF of microscope. Results : Eotaxin mRNA expressed in symptomatic, uncontrolled asthma. Steroid inhibited expression of eotaxin mRNA in asthma. Expression of eotaxin mRNA correlated with eosinophil infiltration in bronchial tissues. Conclusion: Expression of eotaxin mRNA increases in uncontrolled asthma and eotaxin is involved in the recruitment of eosinophils.

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Relationship between the mRNA Expression Levels of Sperm Phospholipids Hydroperoxide Glutathione Peroxidase and Heparin-Binding Protein, and in-Vivo Fertility in Boars (돼지 정자내 Phospholipids Hydroperoxide Glutathione Peroxidase와 Heparin-Binding Protein의 발현 수준과 번식 능력의 관계)

  • Oh, Shin-Ae;Shin, Cha-Gyun;You, Young-Ah;Han, Kyung-Soo;Pang, Myung-Geol
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.133-137
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    • 2007
  • The objective of the study was to determine the relationship between the mRNA expression levels of sperm phospholipids hydroperoxide glutathione peroxidase (PHGPx) and heparin-binding protein (HBP), and in-vivo fertility in boars. The farrowing rate was not correlated with litter size. Sperm PHGPx mRNA expression level of the larger litter size (over 10) group $(2,414.7{\pm}400.7)$ was high that of smaller litter size (below 8) group $(1,875.8{\pm}311.2)$. Sperm HBP mRNA expression level was also higher in the larger litter size $(2,255.9{\pm}360.8)$ group than the smaller litter size $(2,155.4{\pm}378.0)$. However, significant differences were not observed. Sperm PHGPx mRNA level was correlated positively with litter size (r=0.206). Because the expression levels of PHGPx and HBP are not strongly correlated with in-vivo fertility, PHGPx and HBP can not be considered a predictive measure for fertility in boars.

Expression of N-Methylpurine-DNA Glycosylase Gene during Fetal Development and Adult in Mice (생쥐 태아 및 성체 조직에서의 N-Methylpurine-DNA Glycosylase 유전자의 발현)

  • Sohn, Tae-Jong;Kim, Nam-Keun;Lee, Sook-Hwan;Han, Sei-Yul;Ko, Jung-Jae;Park, Chan;Lee, Woo-Sik;Lee, Chan;Lee, Yong-Hee;Cha, Kwang-Yul
    • Development and Reproduction
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    • v.3 no.1
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    • pp.101-105
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    • 1999
  • N-Methylpurine-DNA glycosylase (MPG) removes N-methylpurine and other damaged purines in DNA. RT-PCR analysis revealed MPG mRNA expression at various tissues of fetal development from day 8 to day 18 fetus and day 400 mature adult. The MPG transcripts were abundant during fetal development in mice. In placenta, the MPG mRNA was continuously decreased from day 8 post coitum (p.c) to day 18 p.c. fetus. The high level of mRNA in fetal brain and liver was drastically declined in day 400 mature adult. The expression of MPG, originally characterized by its highest level of expression in the epididymis of adult mouse, was detected with high level in several other reproductive organ, including the ovary, oviduct, testis, vas deference, uterus, and seminal vesicles. These results demonstrate developmental stage- and tissue-specific variation of MPG gene expression.

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Expression of Endometriosis Related Genes in the Shed Endometrial Tissues from Menstrual Blood (생리혈에 존재하는 자궁내막조직에서 자궁내막증 관련 유전자의 발현 양상)

  • Park, Chan-Woo;Jun, Jin-Hyun;Koong, Mi-Kyoung;Song, In-Ok
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.4
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    • pp.275-283
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    • 2007
  • Objective: This study was performed to investigate the expressions of endometriosis related genes in shed endometrial tissues from menstrual blood of patients with or without endometriosis. Methods: The shed endometrial tissues were collected on 2$^{nd}$ or 3$^{rd}$ day of menstrual cycle with Wallace catheter in patients with endometriosis (n=16) and without endometriosis (n=26). The mRNA expressions of twelve kinds of endometriosis related genes were compared between two groups using semi-quantitative RT-PCR. Results: The collected shed endometrium was confirmed by histological observation. Expressions of telomerase, c-kit and aromatase mRNA were not detected by RT-PCR in shed endometrial tissues. The mRNA expressions of apoptosis related genes (fas, fas ligand, bcl-2, bax), stem cell factor, estrogen receptor-$\alpha$/$\alpha$, endometriosis protein-I and secretory leukocyte protease inhibitor gene were similar between shed endometrial tissues with endometriosis and without endometriosis. Conclusion: We could not find the difference of mRNA expressions of tested endometriosis related genes between shed endometrial tissues with or without endometriosis by semi-quantitative RT-PCR analysis. It may be related to the dynamical changes of gene expressions in the endometrium with menstrual cycle.

Correlation Between the Expression of Amino Acid Transporter LAT1 mRNA and the Amount of L - Leucine Transport in Human Cancer Cell Lines (사람 암세포주들에서 아미노산 수송체 LAT1 mRNA 발현과 아미노산 L - Leucine 수송의 상관관계)

  • Kim, Do-Kyung;Song, Soo-Keun;Kim, In-Jin;Kook, Joong-Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1451-1456
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    • 2004
  • Amino acid transporters play an important role in supplying nutrients to normal and cancer cells for cell proliferation. System L is a major transport system responsible for the $Na^+$ -independent, large neutral amino acids including several essential amino acids. L-type amino acid transporter 1 (LAT1), an isoform of system L amino acid transporter, is highly expressed presumably to support their continuous growth and proliferation in malignant tumors. In the present study, we have examined the correlation between the expressions of amino acid transporter LAT1 mRNA and its subunit 4F2hc mRNA and the amount of L-leucine transport in various human cancer cell lines. Northern blot analysis have revealed that the 26 human cancer cell lines expressed LAT1 mRNA and 4F2hc mRNA. There were the differences for the levels of LAT1 and 4F2hc mRNA expressions in the 26 human cancer cell lines. The 26 human cancer cell lines transported the L-[$^{14}C$]leucine into the cells via amino acid transporter. In the 26 human cancer cell lines, a linear relationship was observed between the expression of amino acid transporter LAT1 mRNA and the amount of L-leucine transport. Little relationship was observed between the expression of 4F2hc mRNA and the amount of L-leucine transport, but the statistical significance of difference was not detected. These results indicate that the 26 human cancer cell lines express LAT1 mRNA and 4F2hc mRNA and there is the correlation between the expression of amino acid transporter LAT1 mRNA and the amount of L-leucine transport. In addition, specific inhibition of LAT1 in cancer cells will be a new rationale for anti-cancer therapy.

Expression of Nesfatin-1/NUCB2 and Its Binding Site in Mouse Ovary (생쥐 난소 내 Nesfatin-1/NUCB2 발현과 결합 부위 확인)

  • Kim, Jin-Hee;Youn, Mi-Ra;Bang, So-Young;Sim, Ji-Yeon;Kang, Hee-Rae;Yang, Hyun-Won
    • Development and Reproduction
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    • v.14 no.4
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    • pp.287-295
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    • 2010
  • It was recently reported that nesfatin-1/NUCB2, which is secreted from the brain, controls appetite and energy metabolism. The purpose of this research was to confirm whether or not the protein and its binding site should have been expressed in the mouse reproductive organs and to know the possible effects of nesfatin-1 on the reproductive function. Using the ICR female mouse ovary and uterus, the expression of NUCB2 mRNA was confirmed with the conventional PCR and the relative amount of NUCB2 mRNA in the tissues was analyzed with real-time PCR. Immunohistochemical staining was performed using the nesfatin-1 antibody to investigate the nesfatin-1 protein expression and the biotin conjugated nesfatin-1 to confirm the binding site for nesfatin-1 in the ovary. Furthermore, in order to examine if the expression of NUCB2 mRNA in the ovary and uterus is affected by gonadotropin, its mRNA expression was analyzed after PMSG administration into mice. As a result, the expression level of NUCB2 mRNA in the ovary and the uterus was as much as the expression level in hypothalamus. As a result of the immunohistochemical staining, nesfatin-1 proteins were localized at the theca cells, the interstitial cells, and some of the luteal cells. However, the granulosa cells in the follicles did not stain. Interestingly, the oocytes in the some follicles were stained with nesfatin-1. On the other hand, nesfatin-1 protein binding sites were displayed at the theca cells and the interstitial cells near the tunica albuginea. After PMSG administration the expression level of NUCB2 mRNA was increased in the ovary and the uterus. These results demonstrate that for the first time the nesfatin-1 and its binding site were expressed in the ovary and NUCB2 mRNA expression was controlled by gonadotropin, suggesting an important role in the reproductive organs as a local regulator. Therefore, further study is needed to elucidate the functions of nesfatin-1 on the reproductive organs.