• Title/Summary/Keyword: Tropomyosin

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Molecular Cloning of Novel Genes Specifically Expressed in Snailfish, Liparis tanakae (꼼치, Liparis tanakae에서 특이하게 발현되는 새로운 유전인자의 검색)

  • 송인선;이석근;손진기
    • Development and Reproduction
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    • v.4 no.1
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    • pp.67-77
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    • 2000
  • Snailfish usually lives at the bottom of the sea and showed typical retrogressive change with specialized tissue structures of skin and skeletons. In order to obtain the specific genes of snailfish, highly expressed in the body, we made subtracted cDNA library and analyzed 200 clones. Totally 200 clones were obtained and sequenced, and among them 62 clones were turned out to be homologous to the known gene, i.e., thioesterase (9), myosin (8), creatine kinase (7), skeletal alpha-actin (6), parvalbumin b (5), ribosomal protein (5), type I collagen (3), muscle troponin (3), dopamine receptor (2), histatin (2), and heat shock protein (2), cystatin (1), lectin (1), statherin (1), secretory carrier membrane protein (1), keratin type I (1), desmin (1), chloroplast (1), muscle tropomyosin (1), reticulum calcium ATPase (1), ribonucleoprotein (1). The remaining 138 clones were low homologous or non-redundant genes through Genbank search. Especially 5 clones were novel and specifically expressed in the body tissues of Snailfish by in situ hybridization. Therefore, we analysed these 5 clones to identify the C-terminal protein structures and motifs, and partly defined the roles of these proteins in comparison with the expression patterns by in situ hybridization. C9O-77, about 5000 bp, was supposed to be a matrix protein expressed strongly positive in epithelium, myxoid tissue, fibrous tissue and collagenous tissue. C9O-116, about 1500 bp, was supposed to be a transmembrane protein which was weakly expressed in the fibrous tissue, epithelium tissue, and myxoid tissue, but strong in muscle tissue. C9O-130, about 1200 bp, was supposed to be an intracytoplasmic molecule usually in the epithelial cells. C9O-161, about 2000 bp, was weakly expressed in epithelium, muscle tissue and myxoid tissue, but specially strong in epithelium. C9O-171, about 1000 bp, was supposed to be a transcription factor containing zinc finger like domain, which was intensely expressed in the epithelium, muscle tissue, fibrous tissue, and in collagenous tissue.

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Identification of intrinsic regulators in the secondary palate morphogenesis (이차구개 형태분화의 내적 조절유전자 규명)

  • Lee, Jae-Guk;Jang, Eun-Ha;Im, Yang-Hee;Kim, Ki-Byeung;Ko, Seung-O;Cho, Eui-Sic;Shin, Hyo-Keun
    • Korean Journal of Cleft Lip And Palate
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    • v.10 no.1
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    • pp.1-16
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    • 2007
  • 이차구개는 발생과정에서 구개선반의 형성과 성장, 거상과 융합의 과정을 통해 형성된다. 이와 같은 이차구개의 형성과정은 미세한 분자유전학적 신호전달기전에 의해 조절되는 것으로 알려져 있어서, 신호전달과정에 관여하는 유전인자의 발현이상이 되면 정상적인 이차구개가 형성되지 못하고 구개파열이라는 선천성 기형이 발생된다. 구개파열의 유발인자들에 대한 많은 연구에도 불구하고 현재까지 정상적인 이차구개의 형성을 조절하는 분자유전학적 기전에 대해서는 명확히 알려져 있지 않다. 따라서 본 연구에서는 이차구개의 형태분화를 조절하는 분자유전학적 기전을 알아보고자, 이차구개 형성의 내적 조절인자 중 핵심유전자로 알려져 있는 Osr2가 결손된 생쥐의 이차구개 형성과정에서 정상생쥐에 비해 발현의 변동이 나타나는 유전자를 확인하였다. 유전자 발현의 변동은 발생 14.5일(E14.5)의 구개선반으로부터 추출한 total RNA를 이용하여 ACP-based GeneFishing PCR을 시행하여 확인하였고, 각각의 변동된 유전자를 동정하여 정상생쥐의 이차구개 형성과정에서의 발현양상을 in situ hybridization을 시행하여 확인하였다. 총 120쌍의 primer를 이용한 검색을 통해서 정상생쥐의 구개선반에 비해 mutant에서 발현이 변동된 유전자는 7개가 검출되었고, 이들은 모두 정상생쥐에 비해 mutant에서 발현이 증가되는 것으로 확인되었다. 검출된 유전자는 vimentin(Vim), ${\beta}$-tropomyosin 2(Tpm2), thioredoxin-like 5(Txnl5), procollagen type II alpha 1(Col2a1), Insulin-like growth factor binding protein 7(IGFbp7), Sui 1 homologs(Sui 1), Defender against cell death1(Dad1)이었다. 검출된 유전자를 동정하여 정상생쥐의 구개 형성과정에서의 발현양상을 알아본 결과, Col2a1 을 제외한 유전자들은 모두 E13.5의 구개선반에서 특이적으로 발현되고 있었으나 구개선반이 융합된 E15.5에서는 Vim, Txnl5 그리고 Dad1 만이 봉합선을 따라 발현이 지속되고 있었다. 이상의 결과로 보아 검출된 유전자들은 구개선반의 형태분화과정에서 발현되어 이차구개의 형성과정에 관여할 것으로 여겨진다. 또한 이들은 이차구개 형성의 내적조절인자인 Osr2의 downstream target 으로 구개선반의 성장과 융합과정에 직접적으로 관여하는 유전물질일 것으로 추정된다.

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Early Time Course of Immunopathologic Changes in Coronary Arteries of Heterotopically Transplanted Mouse Heart (이소 생쥐심장이식 모델에서 조기 시간경과에 따른 관상동맥의 면역병리학적인 변화)

  • 이정렬;임홍국;서정욱
    • Journal of Chest Surgery
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    • v.31 no.12
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    • pp.1119-1126
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    • 1998
  • Background: In this study, we investigated the early time course of expression of the major histocompatibility(MHC) antigens, intercellular adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1(VCAM-1), interleukin-6 and the histopathological changes in the coronary arteries of cardiac allografts exchanged between inbred mice strains that differ in one loci of class I major histocompatibility antigen (B10.BR to B10.A). Material and Method: No immunosuppressive therapy was used. Both allografts and the hearts of the recipients were harvested at 7(group 1, n=6), 15(group 2, n=6), 21(group 3, n=6), and 30(group 4, n=6) days after transplantation. They were examined by immunohistochemistry, microscopy and morphometry. All allografts had contractions at the time of harvest. Result: A strong MHC class I antigen expression was present on the endothelial and medial cells of the coronary arteries in group 1 and remained unchanged in the rest of the groups. However, MHC class II reactivity was none or very little at any time. Mild to moderate ICAM-1 expression was observed on the endothelial cells, but not on the medial cells at any time by 30 days. VCAM-1 expression was strong both on the endothelial and medial cells at any time. Moderate degree expression of interleukin-6 was observed from 7 to 30 day specimens. Histopathologically, percentage of affected vessels(vessels with intimal thickening) was less than 10 % in 7 day group and increased up to 50 % at 30 days. Mean percent narrowing of the lumen of the affected vessels revealed less than 20 % at 7 days and 40 % at 30 days. The area occupied by tropomyosin positive cells in the intimal lesion, graded from 0 to 3, showed gradual increase but remained between grade 0 to 1 by 30 days. Medial integrity was also well preserved at any time. Moderate perivascular mononuclear cell infiltration was observed at 7 days and it was progressively increased upto 30 days. Recipients' heart revealed no positive immunopathologic findings. Conclusion: In this study, the early time course of progression of the transplantation vasculopathy was demonstrated in the murine heterotopic heart transplant model.

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Expression Pattern of Skeletal-Muscle Protein Genes and Cloning of Parvalbumin mRNA in Dark-banded Rockfish (Sebastes inermis) (볼락(Sebastes inermis) 근육단백질 유전자의 성장단계별 발현 양상과 parvalbumin 유전자 클로닝)

  • Jang, Yo-Soon
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.1-9
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    • 2011
  • Differentially Expressed Gene (DEG) was obtained from Differential Display Reverse Transcription (DDRT)-PCR using Annealing Control Primer (ACP) to search and clone genes related to developmental stages of Sebastes inermis. By using 120 ACPs, the nucleotide sequences obtained from 16 DEGs showing higher expression in 6-month-old skeletal muscle than 18-month-old ones and from 22 DEGs displaying stronger expression in 18-month-old than 6-month-old were analyzed and BLAST was conducted. The results identified that DEGs shared 69~95% homology with genes of parvalbumin (PVALB), nucleoside diphosphate kinase (NDK) B, tropomyosin (TPM), troponin I (TnI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), muscle-type creatine kinase (CKM2), small EDRK-rich factor 2 (SERF2), adenosine monophosphate deaminase (AMPD), Trimeric intracellular cation channel type A (TRICA), Rho GTPase-activating protein 15 (ARHGAP15), S-formylglutathione hydrolase (Esterase D; ESD), heat shock protein 70 (hsp70), type 1 collagen alpha 2 (COL1A2), glutathione S-transferase, Mid1-interacting protein 1 (Mid1lip1), myosin light chain 1 (MYL1), sarcoplasmic/endoplasmic reticulum calcium ATPase 1B (SERCA1B), and ferritin heavy subunit (FTH1). Expression pattern by developmental stage of DEG14 and PVALB exhibiting strong expression in 6-month-old skeletal muscle was investigated using real time PCR. Expression was reduced as Sebastes inermis grew. Expression of PVALB gene was extremely low after 6 months of age. Expression of CKM2 showed higher expression in 18-month-old skeletal muscle than in 6-month-old muscles, and increased continuously until 4 years old, after which CKM2 expression became gradually reduced. By analysis of tissue-specific expression patterns of DEG, DEG14 was expressed mainly in skeletal muscle, liver, kidney and spleen tissues, whereas PVALB expression was expressed in skeletal muscle and kidney, but not in liver and spleen tissues. CKM2 was expressed in skeletal muscle, kidney, and spleen tissues, but not in liver tissues. PVALB gene was composed of 110 amino acids, which constituted 659 bp nucleotides. The results reported here demonstrate that the expression patterns of parvalbumin and CKM2 could be used as molecular markers for selecting fishes exhibiting fast growth.

Regional Differences of Proteins Expressing in Adipose Depots Isolated from Cows, Steers and Bulls as Identified by a Proteomic Approach

  • Cho, Jin Hyoung;Jeong, Jin Young;Lee, Ra Ham;Park, Mi Na;Kim, Seok-Ho;Park, Seon-Min;Shin, Jae-Cheon;Jeon, Young-Joo;Shim, Jung-Hyun;Choi, Nag-Jin;Seo, Kang Seok;Cho, Young Sik;Kim, MinSeok S.;Ko, Sungho;Seo, Jae-Min;Lee, Seung-Youp;Chae, Jung-Il;Lee, Hyun-Jeong
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.8
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    • pp.1197-1206
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    • 2016
  • Adipose tissue in the loin muscle area of beef cattle as a marbling factor is directly associated with beef quality. To elucidate whether properties of proteins involved in depot specific adipose tissue were sex-dependent, we analyzed protein expression of intramuscular adipose tissue (IMAT) and omental adipose tissue (OMAT) from Hanwoo cows, steers, and bulls of Korean native beef cattle by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomic analysis, quantitative polymerase chain reaction (PCR) and western blot analysis. Two different adipose depots (i.e. intramuscular and omental) were collected from cows (n = 7), steers (n = 7), or bulls (n = 7). LC-MS/MS revealed a total of 55 and 35 proteins in IMAT and OMAT, respectively. Of the 55 proteins identified, 44, 40, and 42 proteins were confirmed to be differentially expressed in IMAT of cows, steers, and bulls, respectively. In OMAT of cows, steers, and bulls, 33, 33, and 22 were confirmed to be differentially expressed, respectively. Tropomyosin (TPM) 1, TPM 2, and TPM3 were subjected to verification by quantitative PCR and western blot analysis in IMAT and OMAT of Hanwoo cows, steers, and bulls as key factors closely associated with muscle development. Both mRNA levels and protein levels of TPM1, TPM2, and TPM3 in IMAT were lower in bulls compared to in cows or steers suggesting that they were positively correlated with marbling score and quality grade. Our results may aid the regulation of marbling development and improvement of meat quality grades in beef cattle.