• 한국발생생물학회지:발생과생식
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• 제13권3호
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• pp.199-205
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• 2009

HSP70 has widely been induced in in vivo hyperthermia conditions in various organisms to study gene regulation and recently neuroprotectve roles of the induced gene expression under varying conditions. We investigated different responses among various tissues in zebrafish under heat shock to evaluate whether spatial and temporal expression pattern of zebrafish (z) hsp70 in transcriptional and translational level under heat shock stress in different brain regions. Heat shock groups were given for 1 h at $37^{\circ}C$ after recovery by transferring the treated animals back to $28^{\circ}C$ for 1, 2 and 24 h for recovery, respectively. Control (CTRL) group was kept at $28^{\circ}C$. At the end of treatments, five animals were collected and used for isolation of total RNAs and peptides from the corresponding tissues. Expression of zhsp70 mRNA showed different patterns in recovery periods in the tissues including the brain, eye, intestines, muscles, heart and testis by RT-PCR. Unlike the RT-PCR analysis, Northern blot analysis demonstrated nearly 30-fold increase in zhsp70 at 1 h heat shock, suggesting that RT-PCR may not be appropriate in unmasking regulation of the time-dependent zhsp70 expression. In the experiment involving different brain regions, the cerebellum showed gradual activation at 1 h to R1h and decreases in R2h and R24h, while the medulla oblongata and optic tectum showed gradual increase at R1h and decrease at R24h, indicating that different brain tissues respond specifically to heat shock in inducing zhsp70 and recovering from the heat shock status. Western blot analysis also demonstrated that the intracellular levels of zHSP70 in three different brain regions including the cerebellum, medulla oblongata and optic tectum are differently induced and recovered to normal state. These results clearly demonstrate that different regions of the body and the brain tissues are responding differently to heat shock in the aspects of its level of expression and speed of recovery.

• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2002년도 제1차워크샵
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• pp.139-152
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• 2002

Gene-shaving(Hastie et al, 2000) is a very useful method to identify a meaningful group of genes when the variation of expression is large. By shaving off the low-correlated genes with the leading principal component, the primary genes with the coherent expression pattern can be identified. Gene-shaving method works well If expression levels are varied enough, but it may not catch the meaningful cluster in low expression level or different expression time even with coherent patterns. The sliding window gene-shaving method which is to apply gene-shaving in each sliding window after hierarchical clustering is to compensate losing a meaningful set of genes whose variation is not large but distinct. The performance to identify expression patterns is compared for the simulated profile data by the different variance and expression level.

• Asian-Australasian Journal of Animal Sciences
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• 제32권1호
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• pp.23-30
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• 2019

Objective: Recent studies have demonstrated that lin-28 homolog B (LIN28B)/miRNA let-7 (let-7) plays a role in the regulation of pubertal onset in mammals. However, the role of LIN28B/let-7 in the onset of ovine puberty remains unknown. We cloned the Duolang sheep Lin28B cDNA sequence, detected the expression change of LIN28B, let-7a and let-7g in hypothalamus, pituitary and ovary tissues at three different pubertal stages. Methods: The reverse transcriptase polymerase chain reaction (RT-PCR) was used to clone the cDNA sequence of LIN28B gene from Duolang sheep and the bioinformatics methods were applied to analyze the amino acid sequence of LIN28B protein. The mRNA expression levels of the LIN28B gene at different pubertal stages were examined by real time RT-PCR. Results: LIN28B cDNA of Duolang sheep was cloned, and two transcripts were obtained. The amino acid sequence of transcript 1 shares 99.60%, 98.78%, and 94.80% identity with those of goat, wild yak and pig, respectively. Strong LIN28B mRNA expression was detected in the hypothalamus, pituitary, ovary, oviduct and uterus, while moderate expression was found in the liver, kidney, spleen and heart, weak expression was observed in the heart. No expression was found in the lungs. Quantitative real-time PCR (QPCR) and western-blot analysis revealed that the LIN28B was highly expressed in the hypothalamus and ovary at prepuberty stages, and this expression significantly decreased from the prepuberty to puberty stages (p<0.05). Markedly increased levels of mRNA expression were detected in the pituitary from prepuberty to puberty (p<0.05) and then significantly decreased from puberty to post-puberty (p<0.05). The expression levels of let-7a and let-7g showed no significant changes among different pubertal stages (p>0.05). Conclusion: These results provided a foundation for determining the functions of LIN28B/let-7 and their role in the onset of sheep puberty.

• Journal of Korean Dental Science
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• 제11권2호
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• pp.62-70
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• 2018

Purpose: Bone marrow has long been a source of primary cells. This study was performed to evaluate the effects of age and sex on the cellular viability and expression of stem cell markers of mRNA and on the protein expression of bone marrow stem cells (BMSCs) derived from healthy donors. Materials and Methods: Stem cells were isolated from human bone marrow and plated in culture plates. The shape of the BMSCs was observed under inverted microscope. Quantitative cellular viability was evaluated using a Cell-Counting Kit-8 assay. The expression of stem cell surface markers was tested and a series of quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot was performed to evaluate the expression in each group. Result: The shapes of the cells at 20s, 30s, and 50s were similar to each other. No significant changes in cellular viability were noted among different age groups or sex groups. The BMSCs expressed CD44, CD73, and CD90 surface markers but did not express CD14 and CD34. There were no noticeable differences in CD surface markers among the different age groups. The expressions of CD surface markers were similar between men and women. No significant differences in the secretion of vascular endothelial growth factors (VEGFs) were noted at Day 3 between different age groups. qRT-PCR regarding the expression showed differences between the age groups. However, Western blot analysis showed a decrease in expression but did not reach statistical significance (P>0.05). Conclusion: This study clearly showed no significant differences in shape, cell viability, expression of stem cell surface markers, or secretion of human VEGF among different age groups. However, western blot analysis showed a tendency of age-related decrease which did not reach statistical significance. Collectively, autologous or allogeneic BMSCs should be meticulously applied to obtain optimal results regarding age and sex.

• Entomological Research
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• 제48권5호
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• pp.390-399
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• 2018

In order to precisely assess gene expression levels, the suitable internal reference genes must be served to quantify real-time reverse transcription polymerase chain reaction (RT-qPCR) data. For armyworm, Mythimna separata, which reference genes are suitable for assessing the level of transcriptional expression of target genes have yet to be explored. In this study, eight common reference genes, including ${\beta}$-actin (${\beta}$-ACT), 18 s ribosomal (18S), 28S ribosomal (28S), glyceraldehyde-3-phosphate (GAPDH), elongation fator-alpha ($EF1{\alpha}$), TATA box binding protein (TBP), ribosomal protein L7 (RPL7), and alpha-tubulin (${\alpha}$-TUB) that in different developmental stages, tissues and insecticide treatments of M. separata were evaluated. To further explore whether these genes were suitable to serve as endogenous controls, three software-based approaches (geNorm, BestKeeper, and NormFinder), the delta Ct method, and one web-based comprehensive tool (RefFinder) were employed to analyze and rank the tested genes. The optimal number of reference genes was determined using the geNorm program, and the suitability of particular reference genes was empirically validated according to normalized HSP70, and MsepCYP321A10 gene expression data. We found that the most suitable reference genes for the different experimental conditions. For developmental stages, 28S/RPL7 were the optimal reference genes, both $RPL7/EF1{\alpha}$ were suitable for experiments of different tissues, whereas for insecticide treatments, $28S/{\alpha}-TUB$ were suitable for normalizations of expression data. In addition, $28S/{\alpha}-TUB$ were the suitable reference genes because they have the most stable expression among different developmental stages, tissues and insecticide treatments. Our work is the first report on reference gene selection in M. separata, and might serve as a precedent for future gene expression studies.

• Animal cells and systems
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• 제15권4호
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• pp.268-273
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• 2011

Transgenic mice expressing green fluorescent protein (GFP) under the control of human glial fibrillary acidic protein promoter (hGFAP) have been utilized for in vivo labeling of astrocytes. Although it has been considered that virtually all astrocytes express GFP in this transgenic mouse, we found that different subsets of GFAP-expressing astrocytes express considerably different levels of GFP in the adult brain. Astrocytes in the spinal cord, the molecular layer of thecerebellum, meninges, white matter, corpus callosum and blood vessels exhibited strong GFP, whereas subsets of astrocytes associated with granule cells in the cerebellum and dentate gyrus did not or only marginally exhibited GFP. We also found that a small subset of GFP-expressing cells in the periglomeruli of the olfactory bulb did not express GFAP immunoreactivity. Collectively, these results suggest that human GFAP promoter-derived GFP expression does not faithfully recapitulate the endogenous GFAP expression in mice, suggesting that upstream regulatory mechanisms controlling GFAP transcription are different in different populations of astrocytes, and may reflect the functional diversity of astrocytes.

• The Plant Pathology Journal
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• 제15권1호
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• pp.8-13
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• 1999

Hypersensitive cell death of plants during incompatible plant-pathogen interactions is one of the efficient defense mechanisms of plants against pathogen infections. For better understanding of the molecular mechanisms involved in the plant hypersensitive response (HR), TMV-induced biotic plant cell death and CuSO4-induced abiotic plant cell death were compared in terms of expression patterns of ten different defense-related genes as molecular markers. The genes include five pathogenesis-related protein genes, two plant secondary metabolite-associated genes, two oxidative stress-related genes and one wound-inducible gene isolated from tobacco. Northern blot analyses revealed that a same set of defense-related genes was induced during both biotic and abiotic cell death but with different time and magnitude. The expression of defense-related genes in tobacco plants was temporarily coincided with the time of cell death. However, when suspension cell cultures was used to monitor the expression of defense-related genes, different patterns of the gene expression were detected. This result implies that three are common and, in addition, also different branches of signaling pathways leading to the induced expression of defense-related genes in tobacco during the pathogen- and heavy metal-induced cell death.

• International Journal of Internet, Broadcasting and Communication
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• 제13권4호
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• pp.11-21
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• 2021

The causes of noise generation according to the classification of indoor spaces are very diverse. Individual happiness is infringed by this noise. In this paper, We tried to visualize the spatial sound characteristics of public places using sound color to express them so that anyone can sympathize. The noise inside a conference room of a medical device company was measured for 100 minutes, and the frequency band was divided into three different types of existing sound pressure expression units. Because the size of the noise is expressed differently depending on the situation, There are cases where there is a difference of opinion between the measurer and the researcher. This noise measurement experiment was conducted, and the sound color was applied to classify it on a log scale considering auditory characteristics. As a result of comparing this with the result expression for different loudness expression units, A specific table in different units yielded almost similar results. In addition, the sound source section for 100 minutes was divided into three analysis sections, the analysis sections were different, and the size of the energy ratio for each analysis section was divided in the form of an envelope. The characteristics of the low-frequency region of the space have a high energy ratio, and the decrease in the energy ratio according to the increase in frequency is constant and regular. You can see that conversations are possible.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.1037-1041
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• 2013

Background: Prognostication of breast cancer using clinico-pathologic variables, although useful, remains imperfect. Recent research has focused on finding new markers of prognosis using gene expression profiling. Panels of proteins assessed by immunohistochemistry might also be useful in this regard. This study focused on Bcl-2 protein expression in triple-negative (TNBC) and non- triple-negative breast cancer (non-TNBC) with correlation to clinico-pathologic variables. Materials and methods: We analyzed Bcl-2 expression in 77 women with primary breast carcinoma divided into two groups; triple-negative and non- triple-negative according to expression of estrogen (ER), progesterone (PR) and human epidermal growth factor receptors (Her2/neu). Bcl-2 expression was assessed in relation to age, histo-pathological subtype, grade, nodal status and tumor size. Results: Bcl-2 was expressed in 74% of triple-negative breast cancers and 70% of non- triple-negative cancers. In TNBC, expression was significantly correlated with invasive ductal subtype, while in non-TNBC it was significantly correlated with age and negative nodal status. In both groups higher Bcl-2 expression associated with favourable prognostic factors in breast cancer, but no significant statistical correlations were found. Conclusions: Frequency of Bcl-2 expression does not differ between TNBC and non-TNBC, but different prognostic factors correlate with Bcl-2 in the two cases.

• BMB Reports
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• 제56권9호
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• pp.514-519
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• 2023

Methyltransferase-like 3 (METTL3), a key component of the m⁶A methyltransferase complex, regulates the splicing, nuclear transport, stability, and translation of its target genes. However, the mechanism underlying the regulation of METTL3 expression by alternative splicing (AS) remains unknown. We analyzed the expression pattern of METTL3 after AS in human tissues and confirmed the expression of an isoform retaining introns 8 and 9 (METTL3-IR). We confirmed the different intracellular localizations of METTL3-IR and METTL3 proteins using immunofluorescence microscopy. Furthermore, the endogenous expression of METTL3-IR at the protein level was different from that at the mRNA level. We found that 3'-UTR generation by intron retention (IR) inhibited the export of METTL3-IR mRNA to the cytoplasm, which in turn suppressed protein expression. To the best of our knowledge, this is the first study to confirm the regulation of METTL3 gene expression by AS, providing evidence that the suppression of METTL3 protein expression by IR is an integral part of the mechanism by which 3'-UTR generation regulates protein expression via inhibition of RNA export to the cytoplasm.