• Title/Summary/Keyword: animal expression method

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The mRNA Expression and Methylation Pattern of Apoptosis-related and Imprinted Genes in Day 35 of Cloned Pig Fetuses

  • Jung, Hyun-Ju;Ko, Yeoung-Gyu;Hwang, Seong-Soo;Im, Gi-Sun;Park, Mi-Rung;Woo, Jae-Seok;Park, Choon-Keun;Seong, Hwan-Hoo
    • Reproductive and Developmental Biology
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    • v.31 no.4
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    • pp.227-233
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    • 2007
  • This study was conducted to examine the mRNA expression of apoptosis-related and imprinted genes and methylation pattern of the differentially methylated region (DMR) of H19 gene in day 35 of SCNT pig fetuses. The day 35 of natural mating (control) or cloned (clone) pig fetuses were recovered from uterus. Endometrium from dam and liver from fetus were obtained, respectively. mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. The Bcl-2 mRNA expression in clone was significantly lower than that of control (p<0.05). The mRNA expression of H19 gene in both endometrium and liver was significantly higher in clone than that of control, respectively (p<0.05). The level of IGF-2 mRNA in liver of clone was significantly lower than that of control (p<0.05), whereas the mRNA expression of IGF2-R gene in liver of clone was significantly higher than that of control (p<0.05). The DMR of H19 was lower methylation pattern in clone than that of control. These results suggest that the aberrant mRNA expression of apoptosis-related and imprinted genes and the lower DMR methylation pattern of imprinted gene may be closely related to the inadequate fetal development of cloned fetus.

Gene Co-Expression Network Analysis of Reproductive Traits in Bovine Genome

  • Lim, Dajeong;Cho, Yong-Min;Lee, Seung-Hwan;Chai, Han-Ha;Kim, Tae-Hun
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.185-192
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    • 2013
  • Many countries have implemented genetic evaluation for fertility traits in recent years. In particular, reproductive trait is a complex trait and need to require a system-level approach for identifying candidate genes related to the trait. To find the candidate gene associated with reproductive trait, we applied a weighted gene co-expression network analysis from expression value of bovine genes. We identified three co-expressed modules associated with reproductive trait from bovine microarray data. Hub genes (ZP4, FHL2 and EGR4) were determined in each module; they were topologically centered with statistically significant value in the gene co-expression network. We were able to find the highly co-expressed gene pairs with a correlation coefficient. Finally, the crucial functions of co-expressed modules were reported from functional enrichment analysis. We suggest that the network-based approach in livestock may an important method for analyzing the complex effects of candidate genes associated with economic traits like reproduction.

EARLY SCREENING OF EXPRESSION OF SV40 DRIVEN LACZ INTRODUCED INTO BOVINE EMBRYOS

  • Nakamura, A.;Okumura, J.;Muramatsu, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.5
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    • pp.449-454
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    • 1995
  • The present study was conducted to assess gene expression of bacterial lacZ driven by the SV40 promoter at early developmental stages of bovine embryos. The lacZ gene was linearized with BamHI digestion and introduced into the pronucleus by microinjection at 20 hrs after the commencement of in vitro fertilization. Intact bovine blastocysts were not stained with X-Gal, suggesting that there is no endogenous beta-galactosidase activity in these blastocysts. In contrast, the bovine blastocyst cells microinjected with the lacZ gene exerted a characteristic greenish-blue color originating from the bacterial beta-galactosidase activity, albeit at a low rate, i.e. 2.1% of the total fertilized oocytes injected. It was concluded, therefore, that the lacZ gene driven by the SV40 promoter could be used for an indirect screening method in which the presence of transgene is evaluated from the product of transgene expression.

Enhanced Anti-inflammatory Effects of γ-irradiated Pig Placenta Extracts

  • Kim, Kang Chang;Heo, Jae Hyeok;Yoon, Jong Kwang;Jang, Yuyeon;Kim, Youn Kyu;Kim, Chang-Kyu;Oh, Yu-Kyung;Kim, Young Bong
    • Food Science of Animal Resources
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    • v.35 no.3
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    • pp.293-298
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    • 2015
  • Porcine placenta extract (PPE) is known to possess anti-inflammatory properties owing to its high concentration of bioactive substances. However, the need to eliminate blood-borne infectious agents while maintaining biological efficacy raises concerns about the optimal method for sterilizing PPE. Therefore, the objective of this study was to compare the effects of the standard pressurized heat (autoclaving) method of sterilization with γ-irradiation on the anti-inflammatory effects of PPE. The anti-inflammatory actions of these two preparations of PPE were evaluated by measuring their inhibitory effects on the production of NO, the expression of iNOS protein, and the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA in lipopolysaccharide-stimulated RAW 264.7 cells. Compared with autoclaved PPE, γ-irradiated PPE showed significantly greater inhibition of NO production and iNOS protein expression, and produced a greater reduction in the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA. These results provide evidence that the sterilization process is crucial in determining the biological activity of PPE, especially its anti-inflammatory activity. Collectively, our data suggest that γ-irradiated PPE acts at the transcriptional level to effectively and potently suppresses the production of NO and the expression of pro-inflammatory cytokines.

Analysis and Design Development of Animal Expression Methods Applied in Kids' Wear (아동의류에 나타난 동물표현 분석과 디자인 개발)

  • Kim, Chanju;Ro, Mikyung
    • Journal of the Korean Society of Costume
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    • v.64 no.8
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    • pp.97-110
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    • 2014
  • Animals, as a part of nature, have much influence on children's emotional development and are one of children's favorite subjects of design. This study analyzes the differences in animal expression methods and types of animals, in relation to gender and item based on a collection of children's clothing and accessories that use animal as their theme. The collection consists of 148 clothing and 75 accessories (total of 223 items), which were collected from on-line shopping sites for children's clothes. Analysis results showed the following observations. Animal expression was categorized into four methods: emblem, illustration, character, and pattern.(The most common method was character, followed by pattern, illustration, and emblem. In relation to gender and clothing item, animal expression methods varied widely:(character was the most popular for) boys' t-shirts and accessories, while girls' bottoms and dresses. As for (animal) types, Mammalia was the most common, followed by birds and insects. Dogs and puppies were popular both genders.(However, differences in preferences existed as) boys liked big and wild animals such as bears, tigers, and foxes, while girls liked small and cute animals such as rabbits, cats, owls, and butterflies. Based on these results, six types of animals (rabbits, elephants, large-beaked bird Toco Toucans, fish, penguins, and bees) were selected as themes for kids' vest designs and, among them, three designs were made.

Characteristics of the Animal Motif Found in Modern Fashion (현대패션에 나타난 동물적 모티브의 특성)

  • Kim, Sun-Young
    • Journal of the Korean Home Economics Association
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    • v.50 no.5
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    • pp.39-49
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    • 2012
  • This study looked into the trend of expression of animal motifs that appears in modern fashion, and the values embedded in them. As to the research method, empirical analysis was performed on the works presented at the four global collections of Paris, Milan, New York and London from 2001S/S to 2010F/W, as well as a literature review. As a result, animal motifs applied in modern fashion appeared in material groups, such as leather or feather. The first group shows a realistic material feel, with artificial leather or fur, expressed in a transformed complex animal motif, with process skill or various methods of expression. The second group is the case of a single motif or reality, mode, and ion pattern. Printing is a generic expression, but patchwork, collage or bead ornament are also used, and expressed in the various forms of pattern: pattern in a single animal, pattern in different animals, and pattern combining both animal and other shape. The third group is a small decorative ornament, including headset or accessory and makeup, which is transformed in an animal-characterized motif into part of clothing components for expression, or to set the real animal to the stage for a performance. The embedded values were also categorized as natural beauty, sensual female beauty, pleasure, and ornamentation, which could be an endless spiritual source, and a breakthrough that enables us to show a creative and new aesthetic for the modern fashion field.

Expression Profiles of Cellular Retinol-binding Protein, Type II (CRBP II) in Erlang Mountainous Chickens

  • Yin, H.D.;Tian, K.;Li, D.Y.;Gilbert, E.R.;Xiao, L.H.;Chen, S.Y.;Wang, Y.;Liu, Y.P.;Zhao, X.L.;Zhu, Q.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.3
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    • pp.310-315
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    • 2014
  • Cellular retinol-binding protein II (CRBP II) belongs to the family of cellular retinol-binding proteins and plays a major role in absorption, transport, and metabolism of vitamin A. In addition, because vitamin A is correlated with reproductive performance, we measured CRBP II mRNA abundance in erlang mountainous chickens by real-time PCR using the relative quantification method. The expression of CRBP II showed a tissue-specific pattern and egg production rate-dependent changes. The expression was very high (p<0.05) in jejunum and liver, intermediate in kidney, ovary, and oviduct, and lowest (p<0.05) in heart, hypothalamus, and pituitary. In the hypothalamus, oviduct, ovary, and pituitary, CRBP II mRNA abundance were correlated to egg production rate, which increased from 12 wk to 32 wk, peaked at 32 wk relative to the other time points, and then decreased from 32 wk to 45 wk. In contrast, the expression of CRBP II mRNA in heart, jejunum, kidney, and liver was not different at any of the ages evaluated in this study. These data may help to understand the genetic basis of vitamin A metabolism, and suggest that CRBP II may be a candidate gene to affect egg production traits in chickens.

Selection of Reference Genes for Gene Expression Studies in Porcine Whole Blood and Peripheral Blood Mononuclear Cells under Polyinosinic:Polycytidylic Acid Stimulation

  • Wang, Jiying;Wang, Yanping;Wang, Huaizhong;Hao, Xiaojing;Wu, Ying;Guo, Jianfeng
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.4
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    • pp.471-478
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    • 2014
  • Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (GAPDH, ACTB, B2M, RPL4, TBP, and PPIA) in samples of whole blood and PBMC with or without poly I:C stimulation. Generally, the six candidate genes performed a similar trend of expression stability in the samples of whole blood and PBMC, but more stably expressed in whole blood than in PBMC. geNorm ranked B2M and PPIA as the best combination for gene expression normalization, while according to NormFinder, TBP was ranked as the most stable reference gene, followed by B2M and PPIA. Comprehensively considering the results from the two programs, we recommended using the geometric mean of the three genes, TBP, PPIA and B2M, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation. Our study is the first detailed survey of the gene expression stability in whole blood and PBMC with or without poly I:C stimulation and should be helpful for investigating the molecular mechanism involved in porcine whole blood and PBMC in response to poly I:C stimulation.

Establishment of Spermatogonial Stem Cells using Total Testicular Cell Culture System in Mouse (정소세포의 체외 혼합배양 방법을 이용한 생쥐 정원 줄기세포 확립)

  • Lee, Won Young;Kim, Hee Chan;Kim, Dong Hoon;Chung, Hak Jae;Park, Jin Ki;Song, Hyuk
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.143-148
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    • 2013
  • Spermatogenesis is initiated from spermatogonial stem cells (SSCs) that has an ability of self-renewal and unipotency to generate differentiating germ cells. The objective of this study is to develop the simple method for derivation of SSCs using non-sorting of both spermatogonia and feeder cells. Simply uncapsulated mouse testes were treated with enzymes followed by surgical mincing, and single cells were cultured in stempro-$34^{TM}$ cell culture media at $37^{\circ}C$. After 5 days of culture, aciniform of SSC colony was observed, and showed a strong alkaline phosphatase activity. Molecular characterization of mouse SSCs showed that most of the mouse SSC markers such as integrin ${\alpha}6$ and ${\beta}1$, CD9 and Stra8. In addition, pluripotency embryonic stem cell (ESC) marker Oct4 were expressed, however Sox2 expression was lowered. Interestingly, expression of SSC markers such as Vasa, Dazl and PLZF were stronger than mouse ESC (mESC). This data suggest that generated mouse SSCs (mSSCs) in this study has at least similar biomarkers expression to mESC and mSSCs derived from other study. Immunocytochemistry using whole mSSC colony also confirmed that mSSCs generated from this study expressed SSC specific biomarkers such as c-kit, Thy1, Vasa and Dazl. In conclusion, mSSCs from 5 days old mouse testes were successfully established without sorting of spermatogonia, and this cells expressed both mESC and SSC specific biomarkers. This simple derivation method for mSSCs may facilitate the study of spermatogenesis.

Differential Expression of PPARγ, FASN, and ACADM Genes in Various Adipose Tissues and Longissimus dorsi Muscle from Yanbian Yellow Cattle and Yan Yellow Cattle

  • Ji, Shuang;Yang, Runjun;Lu, Chunyan;Qiu, Zhengyan;Yan, Changguo;Zhao, Zhihui
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.1
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    • pp.10-18
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    • 2014
  • The objective of this study was to investigate the correlation between cattle breeds and deposit of adipose tissues in different positions and the gene expressions of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), fatty acid synthase (FASN), and Acyl-CoA dehydrogenase (ACADM), which are associated with lipid metabolism and are valuable for understanding the physiology in fat depot and meat quality. Yanbian yellow cattle and Yan yellow cattle reared under the same conditions display different fat proportions in the carcass. To understand this difference, the expression of $PPAR{\gamma}$, FASN, and ACADM in different adipose tissues and longissimus dorsi muscle (LD) in these two breeds were analyzed using the Real-time quantitative polymerase chain reaction method (qRT-PCR). The result showed that $PPAR{\gamma}$ gene expression was significantly higher in adipose tissue than in LD in both breeds. $PPAR{\gamma}$ expression was also higher in abdominal fat, in perirenal fat than in the subcutaneous fat (p<0.05) in Yanbian yellow cattle, and was significantly higher in subcutaneous fat in Yan yellow cattle than that in Yanbian yellow cattle. On the other hand, FASN mRNA expression levels in subcutaneous fat and abdominal fat in Yan yellow cattle were significantly higher than that in Yanbian yellow cattle. Interestingly, ACADM gene shows greater fold changes in LD than in adipose tissues in Yan yellow cattle. Furthermore, the expressions of these three genes in lung, colon, kidney, liver and heart of Yanbian yellow cattle and Yan yellow cattle were also investigated. The results showed that the highest expression levels of $PPAR{\gamma}$ and FASN genes were detected in the lung in both breeds. The expression of ACADM gene in kidney and liver were higher than that in other organs in Yanbian yellow cattle, the comparison was not statistically significant in Yan yellow cattle.