• 생명과학회지
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• 제26권7호
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• pp.758-763
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• 2016

본 연구에서는 C2C12 근육 세포의 분화 과정에 있어 IGF-I이 지방산의 수송을 담당하는 지방산 수송체인 FAT/CD36의 mRNA 및 단백질 발현에 미치는 영향에 대해 알아보았다. 그 결과 근육세포의 분화 과정에 있어 FAT/CD36의 단백질과 mRNA 발현이 분화 시간 의존적으로 유의하게 증가하였으며, IGF-I의 처리에 의해서도 유의하게 조절되었음을 알 수 있었다. 이는 IGF-I이 골격근 세포의 성장 및 분화를 촉진하여 근육 관련 유전자들의 발현을 조절하는 기능뿐만 아니라, 골격근의 주요 에너지원으로 사용되는 지방산의 수송을 담당하는 FAT/CD36의 발현에도 영향을 미친다는 것으로 해석할 수 있겠다. 향후 IGF-I이 골격근 세포에서 FAT/CD36의 발현에 영향을 미침으로써 골격근의 지방산 흡수와 산화율을 조절하는지, 그에 따라 지방대사에 어떠한 영향을 미치는지에 대한 연구가 필요할 것이며, 이와 관련된 신호전달 체계 및 기전에 대한 연구도 진행 되어야 할 것이다.

• Restorative Dentistry and Endodontics
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• 제32권5호
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• pp.459-468
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• 2007

이 연구에서는 법랑모세포 분화과정에서 APin의 기능을 알아보고자 APin-protein microarray를 시행한 후 치아발생과 관련이 있는 MEF2, Aurora kinase A, BMPR-IB와 EF-hand calcium binding protein을 분석하여 다음과 같은 결과를 얻었다. 1 CMV-APin construct를 transfection하여 APin의 과발현을 유도한 경우에는 MEF2와 Aurora kinase A 둘 모두에서 발현이 현저히 감소한 반면에, APin의 발현억제를 유도한 경우에는 둘 모두 변화가 없었다. 2. APin의 과발현을 유도한 경우에는 BMPR-IB와 EF-hand calcium binding protein 모두에서 발현이 크게 증가한 반면, APin을 발현억제 시킨 경우에는 BMPR-IB는 변화가 없었고, EF-hand calcium binding protein은 현저히 감소하였다. 위의 결과들로 보아 APin 단백질은 MEF2, Aurora kinase A, BMPR-IB, EF-hand calcium binding protein과 상호작용하여 법랑모세포의 분화와 석회화 과정 중에 중요한 역할을 하는 것으로 사료된다.

• Journal of Plant Biotechnology
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• 제40권1호
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• pp.9-17
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• 2013

Ran is a small GTP-binding protein that binds and subsequently hydrolyzes GTP. The functions of Ran in nuclear transport and mitotic progression are well conserved in plants and animals. In animal cells, stress treatments cause Ran relocalization and slowing of nuclear transport, but the role of Ran proteins in plant cells exposed to stress is still unclear. We have therefore compared Ran genes from three EST libraries construed from different cell types of sweetpotato and the distribution pattern of Ran ESTs differed according to cell type. We further characterized two IbRan genes. IbRan1 is a specific EST to the suspension cells and leaf libraries, and IbRan2 is specific EST to the root library. IbRan1 showed 94.6 % identity with IbRan2 at the amino acid level, but the C-terminal region of IbRan1 differed from that of IbRan2. These two genes showed tissue-specific differential regulation in wounded tissues. Chilling stress induced a similar expression pattern in both IbRan genes in the leaves and petioles, but they were differently regulated in the roots. Hydrogen peroxide treatment highly stimulated IbRan2 mRNA expression in the leaves and petioles, but had no significant effect on IbRan1 gene expression. These results showed that the transcription of these two IbRan genes responds differentially to abiotic stresses and that they are subjected to tissue-specific regulation. Plant Ran-type small G-proteins are a multigenic family, and the characterization of each Ran genes under various environmental stresses will contribute toward our understanding of the distinctive function of each plant Ran isoform.

• 한국응용과학기술학회지
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• 제31권1호
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• pp.101-112
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• 2014

본 연구는 고콜레스테롤 식이를 섭취한 랫드에서 파리유충 에탄올추출물(Ethanolic extract of fly maggot, EM)의 경구투여가 혈액 지질감소에 미치는 영향을 조사하였다. Sprague-Dawley 수컷 랫드를 이용하여 4 처리구(EM 투여량; 대조군=0, 5.0, 7.0, 9.0 mg/100 g 체중)로 구분해서 6주 동안 진행하였다. EM 투여군은 대조군과 비교할 때 혈청 중성지방, 총콜레스테롤, LDL-C가 유의하게 낮았다(p<0.05). HMG-CoA reductase activity는 대조군과 비교할 때 EM 투여군에서 낮았으나 총스테롤, 중성스테롤 및 담즙산 배설량은 EM 투여군에서 유의하게 높았다(p<0.05). EM의 혈액 콜레스테롤 감소와 관련한 생물학적 작용기작을 규명하기 위해서 고콜레스테롤 식이를 섭취한 랫드에서 유도된 생체유전자 sterol response element binding proteins (SREBPs) 및 the peroxisome proliferator-activated receptors ($PPAR{\alpha}$) 발현을 측정하였다. EM은 고콜레스테롤 식이를 공급받은 랫드의 간에서 SREBP-$1{\alpha}$, SREBP-2 mRNA 발현을 억압함과 동시에 $PPAR{\alpha}$ mRNA 발현을 촉진시키는 것으로 나타났다(p<0.05). 본 연구의 결과는 파리유충 에탄올추출물이 고콜레스테롤 식이를 섭취한 랫드에서 지질대사와 관련한 생화학적 매개변수 및 유전자발현 조절을 통하여 혈액 콜레스테롤을 낮춘다는 새로운 사실을 발견하였다.

• 한국식품과학회지
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• 제46권4호
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• pp.498-504
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• 2014

본 연구에서는 3T3-L1 지방전구세포를 이용하여 Allium속 식물의 하나인 두메부추(ASL) 메탄올 추출물의 ROS 생성 저해 및 지질축적 억제 효과를 확인하고자 하였다. 먼저 ASL 메탄올 추출물 $100-2,000{\mu}g/mL$의 모든 농도에서 유의적인 DPPH 라디칼 소거능을 나타내었으며, ASL 메탄올 추출물을 처리하였을 때 유의적인 세포 독성을 나타내지 않은 $10-100{\mu}g/mL$ 농도에서 실험을 진행하였다. 지방세포 내 ROS 관련 효소와 분화관련 전사인자의 조절로 인한 중성지방 축적 저해 활성을 확인하기 위하여 지방전구세포의 분화를 분화 유도하면서 추출물을 농도별(10, 50 및 $100{\mu}g/mL$)로 처리하였다. 그 결과, ASL 메탄올 추출물은 대조군에 비해 ROS 생성량과 ROS 관련 효소인 G6PDH mRNA 발현을 농도 의존적으로 감소시켰다. 또한 ASL 메탄올 추출물 처리로 인하여 지방세포 내 중성지방 축적이 유의적으로 감소되었으며, adipogenic transcription factor인 SREBP1c, $PPAR{\gamma}$ 및 $C/EBP{\alpha}$ mRNA 발현도 농도 의존적으로 억제되었다. 이상의 결과들로 볼 때, ASL 메탄올 추출물로 인한 ROS 생성 저해와 지질축적 억제는 ROS 생성 및 ROS 관련 유전자의 발현 감소로 인한 지방 생성 주요 전사인자의 유전자 발현 억제로 인한 것으로 보이며, ASL이 항비만에 우수한 효능을 가지는 기능성 식품 소재로서의 개발 가능성이 있을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권11호
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• pp.1551-1558
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• 2008

The mammalian cellular retinoic acid-binding proteins, CRABP-I and CRABP-II, bind retinoic acid which acts as an inducer of differentiation in several biological systems. To investigate a possible role for CRABP-II in bovine adipogenesis, we have cloned bovine CRABP-II cDNA and the coding region for CRABP-I. The predicted amino acid sequences of CRABP-II were highly conserved among several animal species (human, mouse, and rat at 97%, 93%, and 93%, respectively). The expression pattern of bovine CRABP-II was examined in greater details by applying RT-PCR to various bovine tissues. CRABP-II mRNA was expressed in most adipose-containing tissues. Moreover, the expression of CRABP-I and -II mRNA dramatically increased during the differentiation of adipocytes from bovine intramuscular fibroblast-like cells. The effects of retinoic acid on adipocyte differentiation of bovine intramuscular fibroblast-like cells were concentration-dependent. Retinoic acid activated the formation of lipid droplets at a level of 1 nM, whereas inhibition was observed at a level of $1{\mu}M$. CRABP-I gene was up-regulated and CRABP-II gene down-regulated by retinoic acid during adipocyte differentiation. These results suggest that CRABPs may play an important role in the regulation of intracellular retinoic acid concentrations during adipogenesis.

• The Korean Journal of Physiology and Pharmacology
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• 제3권3호
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• pp.329-337
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• 1999

Thyroid hormone-induced cellular dysfunctions may be associated with changes in the intracellular $Ca^{2+}$ concentration. The ryanodine receptor, a $Ca^{2+}$ release channel of the SR, is responsible for the rapid release of $Ca^{2+}$ that activates cardiac muscle contraction. In the excitation-contaction coupling cascade, activation of ryanodine receptors is initiated by the activity of sarcolemmal $Ca^{2+}$ channels, the dihydropyridine receptors. In hyperthyroidism left ventricular contractility and relaxation velocity were increased, whereas these parameters were decreased in hypothyroidism. The mechanisms for these changes have been suggested to include alterations in the expression and/or activity levels of various proteins. In the present study, quantitative changes of ryanodine receptors and the dihydropyridine receptors, and the functional consequences of these changes in various thyroid states were investigated. In hyperthyroid hearts, $[^3H]ryanodine$ binding and ryanodine receptor mRNA levels were increased, but protein levels of ryanodine were not changed significantly. However, the above parameters were markedly decreased in hypothyroid hearts. In case of dihydropyridine receptor, there were a significant increase in the mRNA and protein levels, and [3H]nitrendipine binding, whereas no changes were observed in these parameters of hypothyroid hearts. Our findings indicate that hyperthyroidism is associated with increases in ryanodine receptor and dihydropyridine receptor expression levels, which is well correlated with the ryanodine and dihydropyridine binding. Whereas opposite changes occur in ryanodine receptor of the hypothyroid hearts.

• Biomolecules & Therapeutics
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• 제27권2호
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• pp.201-209
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• 2019

Mixed lineage leukemia proteins (MLL) are the key histone lysine methyltransferases that regulate expression of diverse genes. Aberrant activation of MLL promotes leukemia as well as solid tumors in humans, highlighting the urgent need for the development of an MLL inhibitor. We screened and isolated MLL1-binding ssRNAs using SELEX (${\underline{S}}ystemic$ ${\underline{E}}volution$ of ${\underline{L}}igands$ by ${\underline{E}}xponential$ enrichment) technology. When sequences in sub-libraries were obtained using next-generation sequencing (NGS), the most enriched aptamers-APT1 and APT2-represented about 30% and 26% of sub-library populations, respectively. Motif analysis of the top 50 sequences provided a highly conserved sequence: 5'-A[A/C][C/G][G/U][U/A]ACAGAGGG[U/A]GG[A/C] GAGUGGGU-3'. APT1, APT2, and APT5 embracing this motif generated secondary structures with similar topological characteristics. We found that APT1 and APT2 have a good binding activity and the analysis using mutated aptamer variants showed that the site information in the central region was critical for binding. In vitro enzyme activity assay showed that APT1 and APT2 had MLL1 inhibitory activity. Three-dimensional structure prediction of APT1-MLL1 complex indicates multiple weak interactions formed between MLL1 SET domain and APT1. Our study confirmed that NGS-assisted SELEX is an efficient tool for aptamer screening and that aptamers could be useful in diagnosis and treatment of MLL1-mediated diseases.

• Animal Bioscience
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• 제34권4호
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• pp.751-758
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• 2021

Objective: The objectives of this study were to investigate the relationship between the mRNA expression of adipocyte type fatty acid binding protein (A-FABP) and heart type FABP (H-FABP) in Thai native chicken crossbreeds and evaluate the level of exotic inclusion in native chicken that will improve growth while maintaining its relatively low carcass fat. Methods: The fat deposition traits and mRNA expression of A-FABP and H-FABP were evaluated at 6, 8, 10, and 12 weeks of age in 4 chicken breeds (n = 8/breed/wk) (100% Chee breed [CH] [100% Thai native chicken background], CH male and broiler female [Kaimook e-san1; KM1] [50% CH background], broiler male and KM1 female [Kaimook e-san2; KM2] [25% CH background], and broiler [BR]) using abdominal fat (ABF) and muscular tissues. Results: The BR breed was only evaluated at 6 weeks of age. At week 6, the CH breed had a significantly lower A-FABP expression in ABF and intramuscular fat (IF) compared with the other breeds. At 8 to 12 weeks, the KM2 groups showed significant upregulation (p<0.05) of A-FABP in both ABF and IF compared to the CH and KM1 groups. The expression of H-FABP did not follow any consistent pattern in both ABF and IF across the different ages. Conclusion: Some level of crossbreeding CH chickens can be done to improve growth rate while maintaining their low ABF and IF. The expression level of A-FABP correlate with most fat traits. There was no consistency of H-FABP expression across breed. A-FABPs is involved in fat deposition, genetic markers in these genes could be used in marker assisted studies to select against excessive fat accumulation.

• Animal cells and systems
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• 제3권3호
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• pp.311-317
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• 1999

Prolactin (PRL) was reported to be locally synthesized in many brain areas including the hypothalamus, thalamus (TH) and hippocampus (HIP). In the pituitary lactotrophs, PRL synthesis is dependent upon a pituitary-specific transcription factor, Pit-1. In the present study, we attempted to identify Pit-1 or Pit-1-like protein in brain areas known as the synthetic sites of PRL. Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis showed the same Pit-1 transcripts in brain areas such as the medial basal hypothalamus (MBH), preoptic area (POA), TH, and HIP with the Pit-1 transcripts in the anterior pituitary (AP). Electrophoretic mobility shift assay (EMSA) was run with nuclear protein extracts from brain tissues using a double strand oligomer probe containing a putative Pit-1 binding domain. Shifted bands were found in EMSA results with nuclear proteins from MBH, POA, TH and HIP. Specific binding of the Pit-1-like protein was further confirmed by competition with an unlabeled cold probe. Antisense Pit-1 oligodeoxynucleotide (Pit-1 ODN), which was designed to bind to the Pit-1 translation initiation site and block Pit-1 biosynthesis, was used to test Pit-1 dependent brain PRL transcription. Two nmol of Pit-1 ODN was introduced into the lateral ventricle of a 60-day old male rat brain. RNA blot hybridization and in situ hybridization indicated a decrease of PRL mRNA signals by the treatment of Pit-1 ODN. Taken together, the present study suggests that Pit-1 may play an important role in the transcriptional regulation of local PRL synthesis in the brain.