• Asian-Australasian Journal of Animal Sciences
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• 제20권2호
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• pp.184-193
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• 2007

In mammalian ovary, steroidogenic acute regulatory (StAR) protein mediates the true rate-limiting step of transport of cholesterol from outer to inner mitochondrial membrane. Appropriate expression of StAR gene represents an indispensable component of steroidogenesis and its regulation has been found to be species specific. However, limited information is available regarding StAR gene expression during estrous cycle in buffalo ovary. In the present study, expression, localization and hormonal regulation of StAR mRNA were analyzed by semi-quantitative RT-PCR in buffalo ovary and partial cDNA was cloned. Total RNA was isolated from whole follicles of different sizes, granulosa cells from different size follicles and postovulatory structures like corpus luteum and Corpus albicans. Semi-quantitative RT-PCR analyses showed StAR mRNA expression in the postovulatory structure, corpus luteum. No StAR mRNA was detected in total RNA isolated from whole follicles of different size including the preovulatory follicle (>9 mm in diameter). However, granulosa cells isolated from preovulatory follicles showed the moderate expression of StAR mRNA. To assess the hormonal regulation of StAR mRNA, primary culture of buffalo granulosa cells were treated with FSH (100 ng/ml) alone or along with IGF-I (100 ng/ml) for 12 to 18 h. The abundance of StAR mRNA increased in cells treated with FSH alone or FSH with IGF-I. However, effect of FSH with IGF-I on mRNA expression was found highly significant (p<0.01). In conclusion, differential expression of StAR messages was observed during estrous cycle in buffalo ovary. Also, there was a synergistic action of IGF-I on FSH stimulation of StAR gene.

• The Korean Journal of Physiology and Pharmacology
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• 제12권3호
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• pp.117-123
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• 2008

Although growth associated protein-43 (GAP-43) is known to playa significant role in the regulation of axonal growth and the formation of new neuronal connections in the hippocampus, there is only a few studies on the effects of acute stress on GAP-43 mRNA expression in the hippocampus. Moreover, the effects of repeated citalopram treatment on chronic mild stress (CMS)-induced changes in GAP-43 mRNA expression in the hippocampus have not been explored before. To explore this question, male rats were exposed to acute immobilization stress or CMS. Also, citalopram was given prior to stress everyday during CMS procedures. Acute immobilization stress significantly increased GAP-43 mRNA expression in all subfields of the hippocampus, while CMS significantly decreased GAP-43 mRNA expression in the dentate granule cell layer (GCL). Repeated citalopram treatment decreased GAP-43 mRNA expression in the GCL compared with unstressed controls, but this decrease was not further potentiated by CMS exposure. Similar decreases in GAP-43 mRNA expression were observed in CA1, CA3 and CA4 areas of the hippocampus only after repeated citalopram treatment in CMS-exposed rats. This result indicates that GAP-43 mRNA expression in the hippocampus may differently respond to acute and chronic stress, and that repeated citalopram treatment does not change CMS-induced decreases in GAP-43 mRNA expression in the GCL.

• BMB Reports
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• 제47권11호
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• pp.619-624
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• 2014

Antisense RNA is a type of noncoding RNA (ncRNA) that binds to complementary mRNA sequences and induces gene repression by inhibiting translation or degrading mRNA. Recently, several small ncRNAs (sRNAs) have been identified in Escherichia coli that act as antisense RNA mainly via base pairing with mRNA. The base pairing predominantly leads to gene repression, and in some cases, gene activation. In the current study, we examined how the location of target sites affects sRNA-mediated gene regulation. An efficient antisense RNA expression system was developed, and the effects of antisense RNAs on various target sites in a model mRNA were examined. The target sites of antisense RNAs suppressing gene expression were identified, not only in the translation initiation region (TIR) of mRNA, but also at the junction between the coding region and 3' untranslated region. Surprisingly, an antisense RNA recognizing the upstream region of TIR enhanced gene expression through increasing mRNA stability.

• 운동영양학회지
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• 제17권2호
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• pp.49-55
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• 2013

Our purpose was to investigate the effect of a 5-week high-fat diet on the body weight, blood components [triglycerides (TGs), free fatty acids (FFAs), and glucose], and lipoprotein lipase (LPL) mRNA expression in the skeletal muscles of rats. Body weight increased overtime in experimental and control groups without significant differences. In terms of the blood components, the density of TG was significantly lower in the high-fat diet group compared to the control, whereas FFA and glucose levels were similar in the two groups. Although the levels of glyceraldehyde 3-phosphate dehydrogenase(GAPDH) mRNA expression were similar between the groups, LPL mRNA expression was significantly higher in the high-fat diet group than that in the control group. The results of this study suggest that a high-fat diet enhances LPL mRNA expression, as well as possibly increases fat metabolism. For a better understanding of the relationship between diet patterns and fat metabolism, further analysis of genes related to the fat metabolism is warranted.

• Imaging Science in Dentistry
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• 제33권1호
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• pp.51-57
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• 2003

Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, particularly an the expression of type I collagen and alkaline phosphatase mRNA. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1, 2, 4, and 8 Gy at a dose rate of 5.38 Gy/min using a cesium 137 irradiator. The specimens were then harvested and RNA extraction was carried out at 1 and 3 days after irradiation. The extracted RNA strands were reverse-transcribed and the resulting cDNA fragments were amplified by PCR. Results: The irradiated cells demonstrated a dose-dependent increase in type I collagen mRNA expression relative to the control group, with a maximum level of type I collagen mRNA expression occurring at 8 Gy. The degree of type I collagen mRNA expression increased significantly at 1 day after irradiation, but little differences were found between the control group and at the 3rd day. The amount of alkaline phosphatase mRNA expression increased significantly at land 3 days after irradiation in the 1 Gy exposed group compared with the control group. Conclusion: The amount of type I collagen and alkaline phosphatase mRNA expression increased significantly 1 day after irradiation when compared with the control group.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.8-15
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• 2000

Despite extensive investigation, the mechanisms of immune responses to Candida albicans infection remain poorly understood. Using RT-PCR and Northern blot analysis, this study demonstrates the pattern of IL-6 mRNA expression in thioglycollate-elicited mouse peritoneal macrophages and NIH3T3 fibroblasts (NIH3T3) in response to C. albicans. The expression of IL-6 mRNA was detectable in both cell types. However, IL-10 mRNA was only expressed in the macrophages, and IL-4 mRNA was not expressed in neither of the two cell types. Although the phagocytic function of the macrophages was inhibited by Cytochalasin D, these macrophages could still induce the expression of IL-6mRNA. These findings indicated that the phagocytosis of C. albicans is not pivotal in the induction of IL-6 mRNA expression. A Northern blot analysis was used to investigate the dose effects of C. albicans and time-course kinetics of IL-6 mRNA expression at various time points. IL-6 mRNA was expressed in a dose-independent manner, and was detectable as early as 30min after C. albicans stimulation. It was evenly sustained up to 4h. These results can contribute to understanding the mechanism of IL-6 mRNA expression in macrophages and NIH3T3 cells in response to C, albicans.

• Journal of Korean Dental Science
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• 제6권1호
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• pp.22-26
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• 2013

Purpose: Transglutaminase 2 (TGase 2) is expressed by tumor necrosis factor-${\alpha}$ in various carcinoma. The role of TGase 2 expression in salivary gland tumors is not clear yet. Established slaivary gland tumor (SGT)cell line has been used to study the pathogenesis of salivary gland adenocarcinoma on a cellular level in vitro. The pupose of this study were to examine mRNA expression of TGase 2 in SGT cell line compared to other tumor cell lines, and to apply these results to the pathogenesis of salivary gland tumor. Materials and Methods: After SGT, SCC-15, HN 4, and HeLa tumor cell lines were cultured under preconfl uency, and 3 days after postconfl uency, the cells were harvested for total RNA extraction and cDNA preparation. Result: Reverse transcription polymerase chain reaction for semiquantitative mRNA analysis was done. TGase 2 mRNA expression was not induced by confl uency in all the cell lines. TGase 2 mRNA expression was variable but markedly enhanced in SGT cell line. Conclusion: mRNA expression of TGase 2 should play an important role in the pathogenesis of SGT cell line originated from ductal cell.

• Tuberculosis and Respiratory Diseases
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• 제57권1호
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• pp.25-31
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• 2004

연구배경 : IFN-${\gamma}$는 결핵균에 대한 숙주의 면역학적 방어기전에서 핵심적인 역할을 한다. 그러므로 결핵균 항원들이 IFN-${\gamma}$ 유전자 발현에 미치는 영향을 알아보는 것은 결핵균에 대한 숙주의 방어기전을 밝히고 이를 이용한 백신의 개발에 이용될 수 있을 것이다. 방 법 : 결핵성 흉막염 환자의 흉수에서 얻은 림프구 배양액에 결핵균(H37Rv), PPD, Ag85B, man-LAM, ara-LAM을 첨가하여 자극한 후 림프구의 IFN-${\gamma}$ mRNA의 발현 정도를 역전사 중합효소연쇄반응을 이용하여 비교하였다. 결 과 : 1) 결핵균(H37Rv)이 결핵성 흉수 림프구의 IFN-${\gamma}$ mRNA의 발현을 증가시켰다. 2) 결핵균 항원 중 PPD와 Ag85B는 결핵성 흉수 림프구의 IFN-${\gamma}$ mRNA의 발현을 증가시켰지만 man-LAM은 결핵성 흉수 림프구의 IFN-${\gamma}$ mRNA의 발현을 억제시켰다. 3) LAM 중에서 man-LAM은 용량이 증가함에 따라 결핵성 흉수 림프구의 IFN-${\gamma}$ mRNA의 발현의 억제 정도가 증가하였지만 ara-LAM의 경우 이와 같은 현상이 관찰되지 않았다. 결 론 : 결핵성 흉수 림프구의IFN-${\gamma}$ mRNA의 발현은 PPD와 Ag85B의 자극에 의해 항진되지만 man-LAM의 자극에 의해서는 억제되었다.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.813-818
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• 2002

Expression of monokine induced by IFN-$\gamma$(Mig) mRNA is well-known to strictly depend on Interferon-$\gamma$(IFN-$\gamma$). Lipopolysaccharide (LPS) alone Is weakly effective on Mig mRNA expression in mouse Peritoneal macrophages. This study was undertaken to investigate the synergistic effect of LPS and IFN-$\beta$ on chemokine Mig gene expression in mouse peritoneal macrophages. Although IFN-$\beta$ alone was minimally effective, LPS plus IFN-$\beta$ synergized to produce a high level of Mig mRNh. The synergistic effect of LPS and IFN-$\beta$ (LPS/IFN-$\beta$) on Mig mRNA expression was strain-specific. The most effective synergistic effect of LPS/IFN-$\beta$ on the mRNh expression was found in simultaneous stimulation of LPS/IFN-$\beta$. This synergy was modulated at the level of the gene transcription and was not dependent on a new protein synthesis. Synergistic effect of LPS/IFN-$\beta$ also required the activation of $NF-_KB$. Accordingly, these data suggest that LPS/IFN-$\beta$ synergizes the expression of Mig mRNA through a process that depends on a pretranscriptional level and/or coincident Mig mRNA transcription.

• 한국발생생물학회지:발생과생식
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• 제14권3호
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• pp.207-214
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• 2010

The tremendous changes of uterine endometrium are observed during early pregnancy and protease and their inhibitors are involved in regulation of cell proliferation and remodeling of the tissues through remodeling the extracellular matrix (ECM). Some of the proteases and protease inhibitors have been suspected to a factor in endometrial changes but many parts of their expression profiles and the physiological roles are not uncovered. To evaluate the functional roles of them, in this study the expression profiles of proteases and protease inhibitors were analyzed using real-time quantitative PCR analysis. Mmp9 (matrix metalloproteinase 9) mRNA levels peaked on day 4 at the time of implantation. On the other hand, Ela2 (neutrophil elastase, NE) mRNA levels were peaked on day 2 of pregnancy. Its expression were decreased until day 4 of pregnancy but increased rapidly until day 7 of pregnancy and decreased again. NE inhibitor Slpi (secretory leukocyte protease inhibitor, SLPI) mRNA levels were related with the implantation stage and with the levels of Ela2. At the time of implantation the expression levels of Slpi mRNA were about 5 times higher than the Ela2 mRNA in the uterus. In the implantation stage embryos, Mmp9 specific mRNA was only detected at the blastocyst. On the other hand, the expression level of SLPI was higher than that of the Ela2 mRNA at blastocyst and 4.5 day p.c. embryos. Based on these results it is suggested that MMP9, SLPI, and NE have important physiological role in embryo implantation both in uterus and embryos.