• 제목/요약/키워드: total RNA

검색결과 1,770건 처리시간 0.027초

Regulation of Laminin Chain Gene Expression by Ovaria Steroid Hormones in Uterine Tissues of Ovariectomized Mice

  • Chanseob Shim;Dongho Geum;Park, Donchan
    • Animal cells and systems
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    • 제1권1호
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    • pp.115-121
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    • 1997
  • To precisely analyze the role of ovarian steroids in the regulation of laminin chain gene expression in mouse uterine tissues, the ovariectomized mouse model was used. Ovariectomized mice received a single injection of steroid hormones and total RNA was isolated from whole uterine tissues. Messenger RNA levels of each laminin chain (A, 81, and 82) were determined by competitive RT-peR procedures. Estradiol decreased mRNA levels of laminin 81 chain about two-fold, and 82 chain rather moderately. Estradiol-induced inhibition of laminin 81 and 82 chain mRNA levels were completely blocked by pretreatment with estrogen receptor antagonist tamoxifen. Estriol, a short acting estrogen which cannot induce hyperplastic responses of rodent uterine tissues, also showed an inhibitory effect on 81 and 82 chain mRNA levels, while estrone, an inactive estrogen, failed to influence either 8 chain mRNA levels. Effects of steroids on A chain mRNA level were quite different from those on 8 chains. Laminin A chain mRNA level was slightly increased by estradiol treatment, but negatively affected by progesterone. Progesterone treatment greatly increased both 8 chain mRNA levels, but slightly decreased A chain mRNA level compared to the control. The effect of progesterone on laminin chain-specific mRNA levels was further increased by co-injection of estradiol in a time-dependent manner. Progesterone-induced 81 and 82 chain mRNA transcription was inhibited by RU486, a synthetic anti-progesterone /anti-glucocorticoid. The present study demonstrates for the first time that steroids are able to regulate laminin gene expression in mouse uterine tissues, indicating that steroid-regulated laminin gene expression is involved in uterine growth and probably differentiation.

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Regulation of Lipoprotein Lipase by Fasting in Epididymal and Mesenteric Adipocytes of Rats

  • Lee, Jae Joon;Chung, Chung Soo;Lee, Myung Yul
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권5호
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    • pp.715-722
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    • 2008
  • There are marked variations in the activity of lipoprotein lipase (LPL) among adipose depots. The aim of this study was to compare the mechanisms of 24 h of fasting on LPL regulation between epididymal (EPI) adipocytes and mesenteric (MES) adipocytes in rats. 1-Day fasting consistently decreased activities of heparin-releasable LPL, total extractable LPL and cellular LPL markedly in both EPI and MES fat pads. LPL activity in MES fat pads was relatively lower than in the EPI fat pads. Consistent with data on LPL activity, the levels of expression of LPL mRNA in both nutritional states were lower in MES than EPI adipose tissue and isolated adipocytes. The decreased LPL activity after 1 day of fasting in MES adipocytes was explained mainly by a 50% decrease in the relative abundance of LPL mRNA level and a parallel 50% decrease in relative rate of LPL synthesis. In contrast, fasting of 1 day in EPI adipocytes decreased total LPL activity by 47% but did not affect LPL mRNA level or relative rate of LPL synthesis. A decrease in overall protein synthesis contributed to the decreased LPL activity after 1 day fasting both in EPI and MES adipocytes. In MES adipocytes the decrease in LPL activity, LPL mRNA and LPL synthesis were comparable, but in EPI adipocytes the changes in LPL activity were substantially larger than the changes in LPL mRNA level and LPL synthesis. Therefore, fasting decreased fat cell size, LPL activity, LPL mRNA level and relative rate of LPL synthesis in rats, and these effects were more marked in the MES adipocytes. These results clearly demonstrate the regional variations in the metabolic response of adipose tissue and LPL functions to fasting.

The Detection of Plant Viruses in Korean Ginseng (Panax ginseng) through RNA Sequencing

  • Lee, Hong-Kyu;Kim, So-Yeon;Yang, Hee-Ji;Lee, Da-Som;Kwon, Boram;Lee, Dong-Yun;Oh, Jonghee;Lee, Su-Heon
    • The Plant Pathology Journal
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    • 제36권6호
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    • pp.643-650
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    • 2020
  • Korean ginseng (Panax ginseng) is a dicotyledonous, medicinal, perennial plant belonging to the genus Panax of the family Araliaceae. We investigated the occurrence and incidence of plant viruses in Panax ginseng in Korea. A total of 656 leaf samples were combined into one and total RNA was extracted from the polled sample, using RNA sequencing (RNA-Seq), a metatranscriptome analysis of the plant virome was conducted. The virus present in Panax ginseng was confirmed by reverse transcription polymerase chain reaction (RT-PCR) assay using virus-specific primers. In RNA-Seq data analysis, the multiplication protein of four viral contigs including Aristotelia chilensis virus 1 (AcV1), Turnip mosaic virus (TuMV), Watermelon mosaic virus (WMV), and Tobamovirus multiplication protein were discovered. From our metatranscriptome analysis and RT-PCR assay, TuMV and WMV were detected, whereas the three viruses reported in China such as tomato yellow leaf curl China virus; panax notoginseng virus A; and panax virus Y were not found in this study. The distribution of domestic ginseng viruses seems different from that recorded in China. Overall, this is the first plant virome analysis of Panax ginseng in Korea.

SNP Discovery from Transcriptome of Cashmere Goat Skin

  • Wang, Lele;Zhang, Yanjun;Zhao, Meng;Wang, Ruijun;Su, Rui;Li, Jinquan
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권9호
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    • pp.1235-1243
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    • 2015
  • The goat Capra hircus is one of several economically important livestock in China. Advances in molecular genetics have led to the identification of several single nucleotide variation markers associated with genes affecting economic traits. Validation of single nucleotide variations in a whole-transcriptome sequencing is critical for understanding the information of molecular genetics. In this paper, we aim to develop a large amount of convinced single nucleotide polymorphisms (SNPs) for Cashmere goat through transcriptome sequencing. In this study, the transcriptomes of Cashmere goat skin at four stages were measured using RNA-sequencing and 90% to 92% unique-mapped-reads were obtained from total-mapped-reads. A total of 56,231 putative SNPs distributed among 10,057 genes were identified. The average minor allele frequency of total SNPs was 18%. GO and KEGG pathway analysis were conducted to analyze the genes containing SNPs. Our follow up biological validation revealed that 64% of SNPs were true SNPs. Our results show that RNA-sequencing is a fast and efficient method for identification of a large number of SNPs. This work provides significant genetic resources for further research on Cashmere goats, especially for the high density linkage map construction and genome-wide association studies.

The role of ginseng total saponin in transient receptor potential melastatin type 7 channels

  • Kim, Byung Joo
    • Animal cells and systems
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    • 제16권5호
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    • pp.376-384
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    • 2012
  • Although ginsenosides have a variety of physiologic or pharmacologic functions in various regions, there are only a few reports on the effects of transient receptor potential melastatin 7 (TRPM7) channels. Here, we showed evidence suggesting that TRPM7 channels play an important role in ginseng total saponin (GTS)-mediated cellular injury. The combination techniques of electrophysiology, pharmacological analysis, small interfering RNA (siRNA) method and cell death assays were used. GTS depolarized the resting membrane potentials and decreased the amplitude of pacemaker potentials in cultured interstitial cells of Cajal (ICCs) in gastrointestinal (GI) tract. The TRPM7-like currents in single ICCs and the overexpressing TRPM7 in HEK293 cells were inhibited by GTS. However, GTS had no effect on $Ca^{2+}$-activated $Cl^-$ conductance. GTS inhibited the survival of human gastric (AGS) and brea (MCF-7) adenocarcinoma cells. Also, GTS inhibited the TRPM7-like currents in AGS and MCF-7 cells. The GTS-mediated cytotoxicity was inhibited by TRPM7-specific siRNA. In addition, we showed that overexpression of TRPM7 channels in HEK293 cells was inhibited by GTS. Thus, TRPM7 channels are involved in GTS-mediated cell death in AGS and MCF-7 cells, and these channels may represent a novel target for physiological disorders where GTS plays an important role.

제주 근해에서 채집된 나망곰치, Gymnothorax minor (뱀장어목, 곰치과) 엽상자어의 분자동정 및 형태기재 (Molecular Identification and Morphological Description of Gymnothorax minor (Muraenidae, Anguilliformes) Leptocephali off Jeju Island)

  • 이승종;지환성;김진구
    • 한국어류학회지
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    • 제25권2호
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    • pp.59-64
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    • 2013
  • 한국 제주도에서 채집된 곰치과(Muraenidae) 엽상자어 2개체 (전장 36.2~69.2 mm)를 대상으로 mtDNA 12S rRNA 724 base pair 염기서열을 분석한 결과, 한국 통영에서 채집된 나망곰치(Gymnothorax minor) 성어와 100% 일치하였다. 나망곰치 엽상자어는 전체 근절수가 137~139개, 등지느러미 앞 근절수가 32개, 항문앞 근절수가 82~86개, 전장에 대한 체고가 10% 이상인 형태적 특징을 나타내었다.

Transglutaminase 2 mRNA Expression in Salivary Gland Tumor Cell Line

  • Chun, Yoon Kwon;Lee, Chong Heon
    • 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.

Variations of Gonadotropin Subunits mRNA Levels at Different Stages of Ovarian Development in Masu Salmon, Oncorhynchus masou

  • Kim Dae-Jung;Han Chang-Hee;Aida Katsumi
    • Fisheries and Aquatic Sciences
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    • 제2권2호
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    • pp.176-181
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    • 1999
  • The variations of gene expression and pituitary contents of GTH subunits during the ovarian development of masu salmon, Oncorhynchus masou, were investigated. The pituitary GTHs contents was measured by radioimmunoassays (RIAs) using purified GTH subunits and their antibodies. Pituitary contents of GTH $I\beta$ gradually increased from April through August, and reached the maximum in October. On the other hand, pituitary contents of GTH $II\beta$ remained low until August, but they rapidly increased in October. Total RNAs were prepared from pooled pituitaries and the GTH subunits mRNA in pituitaries was quantified by Northern blot hybridization using masu salmon cDNA probes for each GTH subunit. GTH $I\beta$ mRNA level increased with the progression of ovarian maturity. However, GTH $II\beta$ mRNA was detected only at a more advanced stage, and were extremly high at ovulation. A high levels for GTH a mRNA was detected only at ovulation stage. The synchronous increase in pituitary contents and mRNA levels suggested that ovarian maturity in masu salmon was regulated by both GTH I and GTH II.

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Efficient Isolation of Intact RNA from the Soft Coral Scleronephthya gracillimum $(K\ddot{u}kenthal)$ for Gene Expression Analyses

  • Woo, Seo-Nock;Yum, Seung-Shic;Yoon, Moon-Geun;Kim, Sa-Heung;Lee, Jong-Rak;Kim, Jeong-Ha;Lee, Taek-Kyun
    • Animal cells and systems
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    • 제9권4호
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    • pp.205-209
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    • 2005
  • A rapid, simple and efficient method to extract RNA from the adult polyps of a soft coral, marine cnidarian, Scleronephthya gracillimum $(K\ddot{u}kenthal)$; was developed in this study. The highest yield and purity of RNA was obtained with the lysis solution containing 35 mM EDTA, 0.7 M LiCl, 7.0% SDS, and 200 mM Tris-Cl (pH 9.0). Approximately $40{\mu}g$ of total RNA was extracted from 200 mg of liquid nitrogen-pulverized polyp tissue. The ratio of absorbance at 260 nm and 280 nm ranged from 1.8 to 2.0. The results of the reverse transcription polymerase chain reaction (RTPCR) with ${\beta}-actin$ gene specific primers and Northern blot analysis using the same gene probe revealed that the RNA extracted by our method had high quality, and was sufficient for subsequent molecular biological analyses. This method was effective for RNA extraction from other soft coral species which belong to the genus Dendronephthya.

한우 간 및 등심 조직에서 불포화지방산의 조성비율과 Stearoyl-CoA Desaturase mRNA의 발현 양상 (Relationship between Monounsaturated Fatty Acid Composition and Stearoyl-CoA Desaturase mRNA Level in Hanwoo Liver and Loin Muscle)

  • 이승환;윤두학;황수한;정은영;김언현;이창수
    • Journal of Animal Science and Technology
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    • 제46권1호
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    • pp.7-14
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    • 2004
  • 쇠고기내의 oleic acid(18:1)의 힘량은 고기의 향미와 다즙성에 영향올 미치며, 그 함량이 높으면 혈중 콜레스테롤을 낮출 수 있는 요인이 된다. 본 연구는 한우의 간 조직과 등심 조직에서 불포화지방산의 조성과 불포화지방산 합성효소의 mRNA 발현량의 차이를 비교분석하여 그 상호 관련성에 대해 조사하였다. 단일불포화지방산인 palmitoleic acid(16:1)와 oleic acid (18:1)의 조성은 등심 조직에서 총 지방산의 51%를 차지하고 있었으나, 지방대사가 활발한 간 조직에서는 22%에 지나지 않았다. 반면에 포화지방산인 palmitic acid(16:0) 와 stearic acid (18:0)의 조성비는 등심 조직과 간 조직에서 각각 37%와 58%'를 차지하였다. 이같이 단일불포화지방산의 조성은 조직간 현격한 차이가 나타나며, 조직내 stearoyl-CoA desaturase(SCD) 활성을 나타내는 포화지방산에 대한 불포화지방산의 비율인 불포화도(the desaturation index)를 살펴보면 등심 조직은 간 조직보다 약 3.6배 높았다. SCD는 palmitic acid(16:0) 와 stearic acid(18:0)를 기질로 하여 palmitoleic acid(16:1)와 oleic acid (18:1)로 전환하는 중요한 효소이다. 등심 조직과 간 조직에서 단일 불포화지방산의 조성비 차이가 SCD mRNA 의 발현량과 어떤 관련성을 가지는지를 RT PCR 법을 이용하여 분석되었다. 그 결과, SCD mRNA는 등심 조직에서 간 조직보다 약 3배 많이 발현되었다. 이상과 같이 한우 등섬 조직과 간 조직에서 단일불포화지방산의 조성의 현격한 치아는 SCD 활성지표인 지방산의 불포화도(3.6배 차이)와 SCD mRNA 발현량(3배 차이)과 상호 밀접한 관련성으로 해석할 수 있었다.