• Asian-Australasian Journal of Animal Sciences
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• 제12권5호
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• pp.689-694
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• 1999

This experiment was carried out to isolate the partial bovine (Bos Taurus coreanae) myogenic factor encoding gene, MyoD, using the rat myogenic factor (MyoD) gene sequence and to compare the gene sequence between another myogenic factor (Myf 5) and MyoD gene of the bovine. To make the probe and isolate the MyoD gene, PCR was performed to amplify rat and bovine MyoD gene including exon I, II and intron I. The homology between mouse and bovine MyoD is high; bovine MyoD gene shows 17 different gene sequence region compared to rat MyoD. Among those, two regions have significant differences; one is the exon I part between 2834 and 2850 bp, the other is intron part between 3274 and 3303 bp of the mouse. At this region homology was 40% in the former and 50% in the latter. Homology between bovine MyoD and Myf5 was 83% in the exon 1. Especially exon I in the Myf5 602-617 bp and 651-683 bp have significant differences. These results suggest that MyoD gene have a similar gene structure in mouse and bovine and MyoD and Myf5 of the bovine, at least in part, have a similar expression and activity.

• Asian-Australasian Journal of Animal Sciences
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• 제30권7호
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• pp.1029-1036
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• 2017

Objective: In the livestock industry, the regulatory mechanisms of muscle proliferation and differentiation can be applied to improve traits such as growth and meat production. We investigated the regulatory pathway of MyoD and its role in muscle differentiation in quail myoblast cells. Methods: The MyoD gene was mutated by the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 technology and single cell-derived MyoD mutant sublines were identified to investigate the global regulatory mechanism responsible for muscle differentiation. Results: The mutation efficiency was 73.3% in the mixed population, and from this population we were able to establish two QM7 MyoD knockout subline (MyoD KO QM7#4) through single cell pick-up and expansion. In the undifferentiated condition, paired box 7 expression in MyoD KO QM7#4 cells was not significantly different from regular QM7 (rQM7) cells. During differentiation, however, myotube formation was dramatically repressed in MyoD KO QM7#4 cells. Moreover, myogenic differentiation-specific transcripts and proteins were not expressed in MyoD KO QM7#4 cells even after an extended differentiation period. These results indicate that MyoD is critical for muscle differentiation. Furthermore, we analyzed the global regulatory interactions by RNA sequencing during muscle differentiation. Conclusion: With CRISPR/Cas9-mediated genomic editing, single cell-derived sublines with a specific knockout gene can be adapted to various aspects of basic research as well as in functional genomics studies.

• 생명과학회지
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• 제19권10호
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• pp.1456-1462
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• 2009

myo-Inositol의 이성질체인 D-chiro-inositol은 제 2형 당뇨병에 탁월한 효과를 나타내고 있다. D-chiro-inositol의 methylated 형태인 D-pinotol과 D-chiro-inositol은 식물체가 가뭄, 염, 저온과 같은 비 생물학적 스트레스에 노출되게 되면 삼투보호물질로써 합성되어 축적되어 진다. 대두에서는 myo-inositol O-methyltrasnferase에 의하여 ononitol이 합성되고 ononitol epimerase에 의하여 D-pinitol이 합성되고 최종적으로 demethylase에 의하여 D-chiro-inositol이 합성되어 진다. 그러나 쓴메밀에서는 myo-inositol이 직접적으로 D-chiro-inositol로 합성되어 진다는 보고가 있다. 본 실험에서는 쓴메밀에 비 생물학적 스트레스를 처리한 후 cyclitols의 변화를 측정하였다. 그 결과 쓴메밀에서는 myo-inositol epimerase에 의하여 myo-inositol이 D-chiro-inositol로 직접 전환된다는 사실을 확인하였다.

• 대한시과학회지
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• 제20권4호
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• pp.569-577
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• 2018

목적 : 본 연구는 부모의 근시정도에 따라 MC lens, MyoVisonlens 그리고 Single Vision lens를 착용한 그룹들이 각각 어느 정도 효과를 보이는지를 알아보고자 하였다. 방법 : 연구대상은 인천광역시 소재의 한 안경원의 고객 중 2010년 1월에서 2016년 12월 사이에 방문한 7세에서 20세 사이 안질환이 없는 근시안 MyoVision 152안, MC lens 86안, Single Vision lens 270안을 대상으로 등가 구면도수의 변화를 관찰하였다. 본 연구 자료는 MyoVision, MC lens 그리고 Single Vision의 1년간 평균값 변화를 SPSS ver18을 이용하여 분석하였다.각 그룹별로 처음 방문했을 때와 1년 후 재방문 했을 때의 그룹 내 차이를 Paired T-test 이용하여 비교하였고, 이후 One-Way ANOVA(Post Hoc; Bonferroni)분석을 통해 사후분석을 진행하였다. 결과 : 그룹간 비교에서는 MyoVision과 MC lens가 Single Vision 보다 근시 완화효과가 있는 것으로 나타났다. 특히 부모의 굴절이상도에 따라 MyoVision과 MC lens의 근시 완화효과가 다르게 나타났다. 부모 모두 굴절 이상도가 정상일 때는 MyoVision 과 Single Vision lens 간의 변화는 $-0.35{\pm}0.05D$로 통계적으로 유의한 변화값이 나타났다. 아버지만 굴절이상이 있을 때는 MC lens는 Single Vision 보다 $-0.36{\pm}0.14D$만큼 더 효과적인 것으로 나타났다. 그리고 어머니만 굴절이상이 있을 때는 MyoVision 과 Single Vision lens 간의 평균값은 $-0.37{\pm}0.06D$의 변화를 보였고 MC lens 와 Single Vision lens 간의 평균값은 $-0.38{\pm}0.08D$가 변화하여 각각 Single Vision lens에 비해서 근시를 완화시켜주는 효과가 있는 것으로 나타났다.부모모두 굴절이상이 있을 때는 MyoVision 과 Single Vision lens 간의 평균값 변화와 MC lens 와 Single Vision lens 간의 평균값 변화는 각각 $-0.28{\pm}0.07D$, $-0.31{\pm}0.07D$로 MyoVision 과 MC렌즈 모두 통계적으로 유희한 변화값을 보여 근시진행에 억제 효과가 있는 것으로 나타났다. 결론 : 본 연구결과 각 그룹 내에서 MyoVision, MC lens 그리고 Single Vision모두 근시를 완화시켜주는 기능에 있어서 효과가 없는 것으로 보였으나 그룹 간 비교에서는 MyoVision과 MC lens가 Single Vision 보다 근시 완화효과가 있는 것으로 나타났다.

• 대한한의학회지
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• 제27권3호
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• pp.26-37
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• 2006

Determination and differentiation of cells in the skeletal muscle lineage is positively regulated by cell-cell contact. Differentiation proteins proposed to mediate this effect include both classical MyoD and MEF members; potential interactions between the promyogenic activities of these classes of protein, however, are unknown. We show here that MyoD and MEF, two promyogenic family members that relate to each other in a cis fashion, form interactions with MyoD and MEF. These proteins contain myosin-heavy chainsand are enriched at sites of cell-cell contact between myoblasts. Therefore, in differentiation of MyoD and MEF from Chungsimyeonjaeum interact dependently, suggesting that the interactions occur in a cis fashion; consistent with this conclusion, MyoD-mediated differentiation is required for myoblasts to occur by Chungsimyeonjaeum. Inhibition in myoblasts of a MyoD by Staurosporine in its ability to associate with MEF interferes with differentiation as assessed by morphological and transcription levels, suggesting that this interaction is functionally important in myogenesis. Also, some of the differentiation-mediated proteins that are required for myogenesis seem to be based on interdependent activities of the promyogenic classical smad-subfamily.

• BMB Reports
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• 제40권1호
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• pp.22-28
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• 2007

MyoD, expressed in skeletal muscle lineages of vertebrate embryo, is one of muscle-specific basic helix-loop-helix (bHLH) transcription factors, which plays a key role in the determination and differentiation of all skeletal muscle lineages. In this study, a cDNA of grass carp MyoD was cloned and characterized from total RNA of grass carp embryos by RT-PCR. The full-length cDNA of grass carp MyoD is 1597 bp. The cDNA sequence analysis reveals an open reading frame of 825 bp coding for a protein of 275 amino acids, which includes a bHLH domain composed of basic domain (1-84th amino acids) and HLH domain (98-142th amino acids), without signal peptide. Then the MyoD cDNA of grass carp was cloned to yeast expression vector pPICZ$\alpha$A and transformed into P. pastoris GS115 strain, the recombinant MyoD protein with a molecular weight of about 31KD was obtained after inducing for 2d with 0.5% methanol in pH 8.0 BMGY medium, and the maximum yield was about 250 mg/L in shaking-flask fermentation. The results were expected to benefit for further studies on the crystal structure and physiological function of fish MyoD.

• Molecules and Cells
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• 제38권4호
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• pp.362-372
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• 2015

Setdb1, an H3-K9 specific histone methyltransferase, is associated with transcriptional silencing of euchromatic genes through chromatin modification. Functions of Setdb1 during development have been extensively studied in embryonic and mesenchymal stem cells as well as neurogenic progenitor cells. But the role of Sedtdb1 in myogenic differentiation remains unknown. In this study, we report that Setdb1 is required for myogenic potential of C2C12 myoblast cells through maintaining the expressions of MyoD and muscle-specific genes. We find that reduced Setdb1 expression in C2C12 myoblast cells severely delayed differentiation of C2C12 myoblast cells, whereas exogenous Setdb1 expression had little effect on. Gene expression profiling analysis using oligonucleotide microarray and RNA-Seq technologies demonstrated that depletion of Setdb1 results in downregulation of MyoD as well as the components of muscle fiber in proliferating C2C12 cells. In addition, exogenous expression of MyoD reversed transcriptional repression of MyoD promoter-driven luciferase reporter by Setdb1 shRNA and rescued myogenic differentiation of C2C12 myoblast cells depleted of endogenous Setdb1. Taken together, these results provide new insights into how levels of key myogenic regulators are maintained prior to induction of differentiation.

• 대한한방내과학회지
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• 제28권2호
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• pp.333-345
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• 2007

Determination and differentiation of cells in the skeletal muscle lineage is positively regulated by cell-cell contact. differentiation proteins proposed to mediate this effect include both classical MyoD and MEF members : potential interactions between the promyogenic activities of these classes of protein, however, are unknown. We show here that MyoD and MEF, two promyogenic family members that determine to each other in a cis fashion, form ineraction with MyoD- and MEF. These proteins contain myosin heavy chains and are enriched at sites of cell-cell contact between myoblasts, Therefore, In differentiation of MyoD MEF from CST (Chungsimyeonjatang) interact dependently, suggesting that the interactions occur in a cis fashio : consistent with this conclusion, MyoD-mediated differentiation is required for myoblast to occur by CST. Inhibition in myoblasts of a MyoD by STP in its ability to associate with MEF interferes with differentiation as assessed by morphological and transcription level, suggesting that this interaction is functionally important in myogenesis. Also, some of the differentiation-mediated proteins that are required for myogenesis seem to be based on interdependent activities of promyogenic classical SMAD-subfamilly.

• Molecules and Cells
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• 제21권2호
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• pp.206-212
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• 2006

We have established in culture a spontaneously immortalized bovine embryonic fibroblast (BEF) cell line that has lost p53 and $p16^{INK4a}$ functions. MyoD is a muscle-specific regulator capable of inducing myogenesis in a number of cell types. When the BEF cells were transduced with MyoD they differentiated efficiently to desmin-positive myofibers in the presence of 2% horse serum and 1.7 nM insulin. The myogenic differentiation of this cell line was more rapid and obvious than that of C2C12 cells, as judged by morphological changes and expression of various muscle regulatory factors. To confirm that lack of the p53 and $p16^{INK4a}$ pathway does not prevent MyoD-mediated myogenesis, we established a cell line transformed with SV40LT (BEFV) and introduced MyoD into it. In the presence of 2% horse serum and 1.7 nM insulin, the MyoD-transduced BEFV cells differentiated like the MyoD-transduced BEFS cells, and displayed a similar pattern of expression of muscle regulatory proteins. Taken together, our results indicate that MyoD overexpression overcomes the defect in muscle differentiation associated with immortalization and cell transformation caused by the loss of p53 and Rb functions.

• BMB Reports
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• 제40권6호
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• pp.921-927
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• 2007

UCP2 and UCP3 are members of the uncoupling protein family, which may play roles in energy homeostasis. In order to determine the regulation of the predominant expression of UCP3 in skeletal muscle, the effects of differentiation and myogenic regulatory factors on the promoter activities of the mouse UCP2 and UCP3 genes were studied. Reporter plasmids, containing approximately 3 kb of the 5'-upstream region of the mouse UCP2 and UCP3 genes, were transfected into C2C12 myoblasts, which were then induced to differentiate. Differentiation positively induced the reporter expression about 20-fold via the UCP3 promoter, but by only 2-fold via the UCP2 promoter. C2C12 myoblasts were cotransfected with expression vectors for myogenin and/or MyoD as well as reporter constructs. The simultaneous expression of myogenin and MyoD caused an additional 20-fold increase in the reporter expression via the UCP3 promoter, but only a weak effect via the UCP2 promoter. In L6 myoblasts, only MyoD activated the UCP3 promoter, but in 3T3-L1 cells neither factor activated the UCP3 promoter, indicating that additional cofactors are required, which are present only in C2C12 myoblasts. The expression of UCP2 and UCP3 is differentially regulated during muscle differentiation due to the different responsiveness of their promoter regions to myogenin and MyoD.