• Title/Summary/Keyword: MYO

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Efficient transgene expression system using a cumate-inducible promoter and Cre-loxP recombination in avian cells

  • Park, Tae Sub;Kim, Si Won;Lee, Jeong Hyo
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.6
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    • pp.886-892
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    • 2017
  • Objective: Transgenic technology is widely used for industrial applications and basic research. Systems that allow for genetic modification play a crucial role in biotechnology for a number of purposes, including the functional analysis of specific genes and the production of exogenous proteins. In this study, we examined and verified the cumate-inducible transgene expression system in chicken DF1 and quail QM7 cells, as well as loxP element-mediated transgene recombination using Cre recombinase in DF1 cells. Methods: After stable transfer of the transgene with piggyBac transposon and transposase, transgene expression was induced by an appropriate concentration of cumate. Additionally, we showed that the transgene can be replaced with additional transgenes by co-transfection with the Cre recombinase expression vector. Results: In the cumate-GFP DF1 and QM7 cells, green fluorescent protein (GFP) expression was repressed in the off state in the absence of cumate, and the GFP transgene expression was successfully induced in the presence of cumate. In the cumate-MyoD DF1 cells, MyoD transgene expression was induced by cumate, and the genes controlled by MyoD were upregulated according to the number of days in culture. Additionally, for the translocation experiments, a stable enhanced green fluorescent protein (eGFP)-expressing DF1 cell line transfected with the loxP66-eGFP-loxP71 vector was established, and DsRed-positive and eGFP-negative cells were observed after 14 days of co-transfection with the DsRed transgene and Cre recombinase indicating that the eGFP transgene was excised, and the DsRed transgene was replaced by Cre recombination. Conclusion: Transgene induction or replacement cassette systems in avian cells can be applied in functional genomics studies of specific genes and adapted further for efficient generation of transgenic poultry to modulate target gene expression.

TESTING OF NEUROPEPTIDE Y INVOLVEMENT IN BLOOD FLOW REGULATION IN THE FELINE DENTAL PULP USING D-MYO-INOSITOL-1,2,6-TRISPHOSPHATE (D-myo-inositol-1,2,6-trisphosphate를 이용한 neuropeptide Y의 치수혈류 조절기능 연구)

  • Kim, Sung-Kyo
    • Restorative Dentistry and Endodontics
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    • v.21 no.1
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    • pp.366-374
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    • 1996
  • The purpose of this study was to investigate whether D-myo-inositol-l,2,6-trisphosphate (PP56) can effectively antagonize vasoconstriction caused by neuropeptide Y in the dental pulp, and to understand involvement of neuropeptide Y in the regulation of microcirculation in the dental pulp with the aim of elucidating neurogenic inflammation. Experiments were performed on 7 cats anesthetised with sodium pentobarbital, and neuropeptide Y and a neuropeptide Y antagonist PP56 were injected close intra-arterially into the dental pulp. The probe of laser Doppler flowmeter was placed on the buccal surface of ipsilateral canine teeth to the drug administration and pulpal blood flow was measured. Intra-arterial injection of neuropeptide Y (1.3-$2.0\;{\mu}g$/kg) resulted in pulpal blood flow decrease of $37.73{\pm}5.73%$(mean${\pm}$SEM) (n=9). Intra-arterial injection of PP56(0.3 mg/kg) alone changed pulpal blood flow little by 1.03 % reduction. The effect of neuropeptide Y in the presence of PP56 resulted in significantly less decreases in pulpal blood flow ranging from $27.17{\pm}5.37$ to $16.63{\pm}3.48%$ from control as compared with neuropeptide Y alone(n = 13). In effect, PP56 attenuated pulpal blood flow caused by neuropeptide Y. Results of the present study have provided evidences that a non-peptide PP56 is capable of antagonizing vasoconstriction caused by neuropeptide Y in the feline dental pulp. In addition, they show functional evidences that neuropeptide Y plays an active role in modulating the microcirculation of the dental pulp.

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Effect of Low-Energy Laser Irradiation on the Proliferation and Gene Expression of Myoblast Cells (저출력 레이져 자극이 근육세포의 증식 및 유전자 발현에 미치는 효과)

  • Kwag, J.H.;Jeon, O.H.;Kang, D.Y.;Ryu, H.H.;Kim, K.H.;Jung, B.J.;Kim, C.H.
    • Journal of Biomedical Engineering Research
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    • v.31 no.1
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    • pp.81-86
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    • 2010
  • Laser irradiation is known to affect various tissues such as skin, bone, nerve, and skeletal muscle. Laser irradiation promotes ATP synthesis, facilitates wound healing, and stimulates cell proliferation and angiogenesis. In skeletal muscle, laser irradiation is related to the proliferation of skeletal muscle satellite cells. Normal skeletal muscle contains remodeling capacity from myogenic cells that are derived from mononuclear satellite cells. Their processes are activated by the expression of genes related with myogenesis such as muscle-specific transcription factors (MyoD and Myf5) and VEGF (vascular endothelial growth factor). In this study, we hypothesized that laser irradiation would enhance and regulate muscle cell proliferation and regeneration through modulation of the gene expressions related with the differentiation of skeletal muscle satellite cells. $C_2C_{12}$ myoblastic cells were exposed to continuous/non-continuous laser irradiation (660nm/808nm) for 10 minutes daily for either 1 day or 5 days. After laser irradiation, cell proliferation and gene expression (MyoD, Myf5, VEGF) were quantified. Continuous 660nm laser irradiation significantly increased cell proliferation and gene expression compared to control, continuous 808nm laser irradiation, and non-continuous 660nm laser irradiation groups. These results indicate that continuous 660nm laser irradiation can be applied to the treatment and regeneration of skeletal muscle tissue.

Inhibitory effects of Sam-Myo-San on the LPS-induced production of nitric oxide and $TNF-{\alpha}$ in RAW 264.7 cells and BV-2 Microglia cells (삼묘환(三妙丸)의 LPS에 의해 활성화된 RAW 264.7 cells과 BV-2 Microglia cells로부터 생성되는 nitric oxide 및 $TNF-{\alpha}$의 생성억제효과)

  • Lee, Jae-Hyun;Jung, Hyo-Won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.59-67
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    • 2006
  • Objectives : Sam-Myo-Whan(SMW) has been known traditional prescription with anti- anthritis activities. We investigated inhibitory effects of SMW on lipopolysaccharide (LPS)-induced nitric oxide(NO), $TNF-{\alpha}$ and inducible nitric oxide synthase(iNOS) production from RAW264.7 cells and BV-2 Microglia cells. Methods : SMW, which had been extracted with 70% MeOH, concentrated and freeze-dried was used for this experiment. After BV2 mouse brain macrophages and RAW264.7 mouse peritoneal macrophages were pretreated with increasing concentrations of SMW extract for 30min, and then activated with LPS. To investigate cytotoxicity of SMW extract, cell viability was measured by MTT assay. NO production was measured in each culture supernatant by Griess reaction. mRNA expression of iNOS in two type cells was investigated by RT-PCR. $TNF-{\alpha}$ production was measured in each culture supernatant by ELISA. Results : SMW extract significantly inhibited LPS-induced NO and $TNF-{\alpha}$ production in BV2 cells and RAW264.7 cells dose-dependently. SMW extract also greatly suppressed mRNA expression of iNOS in both type cells activated with LPS. Conclusion : These data suggests that SMW extract may have an anti-inflammatory effect through the inhibition of iNOS expression.

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Effect of Antioxidant Fortification on Preservability of Buffalo Semen

  • Raina, V.S.;Gupta, A.K.;Singh, Kiran
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.1
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    • pp.16-18
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    • 2002
  • During the process of freezing, spermatozoa suffer cold shock which increases their susceptibility to lipid peroxidation which plays an important role in ageing of spermatozoa, shortening their life span and affecting the preservation of semen. An experiment was therefore conducted to study the effect of addition of natural antioxidants into semen diluents on the preservability of buffalo semen. Split semen samples were extended in milk egg yolk diluents fortified with vitamin E (MYE), vitamin C (MYC) and control group (MYO); Tris-egg yolk diluents fortified with vitamin E (TYE), vitamin C (TYC) and control group (TYO) and evaluated for their preservabilities at 4-7$^{\circ}C$ and $37^{\circ}C$. Overall least squares mean of percent motility observed after 0, 24, 48, 72 and 96 h of preservation at 4-7$^{\circ}C$ were 66.70, 54.00, 36.80, 21.90 and 12.50, respectively while the estimates for semen extended in MYE, MYC, MYO, TYE, TYC and TYO were 44.80, 42.70, 38.70, 36.00, 35.20 and 33.00 percent, respectively. The results showed that motility was significantly (p<0.01) affected by extender (extender-antioxidant combination) and preservation interval. Overall least squares mean percent motility observed after 0, 4, 8, 12 and 24 h of preservation at $37^{\circ}C$ were 68.50, 58.90, 45.00, 38.10 and 18.10 percent, respectively, while the estimates for semen extended in MYE, MYC, MYO, TYE, TYC and TYO were 48.20, 49.30, 46.80, 45.30, 42.30 and 42.50 percent, respectively. Extender and storage interval were found to be significantly (p<0.01) affecting spermatozoa motility on room temperature preservation. The results indicated that the incorporation of antioxidants, especially vitamin E, had beneficial effect on preservability of buffalo semen.