• Reproductive and Developmental Biology
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• 제37권4호
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• pp.219-224
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• 2013

Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90~100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson's, oil red O, and Alizarin red staining respectively. We performed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteoblast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were induced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strategies for augmenting meat quality.

• Molecules and Cells
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• 제44권10호
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• pp.770-779
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• 2021

Transgenic Arabidopsis thaliana expressing an anti-rabies monoclonal antibody (mAb), SO57, was obtained using Agrobacterium-mediated floral dip transformation. The endoplasmic reticulum (ER) retention signal Lys-Asp-Glu-Leu (KDEL) was tagged to the C-terminus of the anti-rabies mAb heavy chain to localize the mAb to the ER and enhance its accumulation. When the inaccurately folded proteins accumulated in the ER exceed its storage capacity, it results in stress that can affect plant development and growth. We generated T₁ transformants and obtained homozygous T₃ seeds from transgenic Arabidopsis to investigate the effect of KDEL on plant growth. The germination rate did not significantly differ between plants expressing mAb SO57 without KDEL (SO plant) and mAb SO57 with KDEL (SOK plant). The primary roots of SOK agar media grown plants were slightly shorter than those of SO plants. Transcriptomic analysis showed that expression of all 11 ER stress-related genes were not significantly changed in SOK plants relative to SO plants. SOK plants showed approximately three-fold higher mAb expression levels than those of SO plants. Consequently, the purified mAb amount per unit of SOK plant biomass was approximately three times higher than that of SO plants. A neutralization assay revealed that both plants exhibited efficient rapid fluorescent focus inhibition test values against the rabies virus relative to commercially available human rabies immunoglobulins. KDEL did not upregulate ER stress-related genes; therefore, the enhanced production of the mAb did not affect plant growth. Thus, KDEL fusion is recommended for enhancing mAb production in plant systems.

• Animal Bioscience
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• 제34권6호
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• pp.1078-1087
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• 2021

Objective: Skeletal muscle satellite cells (SMSCs) are significant for the growth, regeneration, and maintenance of skeletal muscle after birth. However, currently, few studies have been performed on the isolation, culture and inducing differentiation of goose muscle satellite cells. Previous studies have shown that C1q and tumor necrosis factor-related protein 3 (CTRP3) participated in the process of muscle growth and development, but its role in the goose skeletal muscle development is not yet clear. This study aimed to isolate, culture, and identify the goose SMSCs in vitro. Additionally, to explore the function of CTRP3 in goose SMSCs. Methods: Goose SMSCs were isolated using 0.25% trypsin from leg muscle (LM) of 15 to 20 day fertilized goose eggs. Cell differentiation was induced by transferring the cells to differentiation medium with 2% horse serum and 1% penicillin streptomycin. Immunofluorescence staining of Desmin and Pax7 was used to identify goose SMSCs. Quantitative realtime polymerase chain reaction and western blot were applied to explore developmental expression profile of CTRP3 in LM and the regulation of CTRP3 on myosin heavy chains (MyHC), myogenin (MyoG) expression and Notch signaling pathway related genes expression. Results: The goose SMSCs were successfully isolated and cultured. The expression of Pax7 and Desmin were observed in the isolated cells. The expression of CTRP3 decreased significantly during leg muscle development. Overexpression of CTRP3 could enhance the expression of two myogenic differentiation marker genes, MyHC and MyoG. But knockdown of CTRP3 suppressed their expression. Furthermore, CTRP3 could repress the mRNA level of Notch signaling pathway-related genes, notch receptor 1, notch receptor 2 and hairy/enhancer-of-split related with YRPW motif 1, which previously showed a negative regulation in myoblast differentiation. Conclusion: These findings provide a useful cell model for the future research on goose muscle development and suggest that CTRP3 may play an essential role in skeletal muscle growth of goose.

• Journal of Microbiology and Biotechnology
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• 제29권7호
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• pp.1165-1176
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• 2019

Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are the most toxic substances known. However, the number of currently approved medical countermeasures for these toxins is very limited. Therefore, studies on therapeutic antitoxins are essential to prepare for toxin-related emergencies. Currently, more than 10,000 Halla horses, a crossbreed between the native Jeju and Thoroughbred horses, are being raised in Jeju Island of Korea. They can be used for equine antitoxin experiments and production of hyperimmune serum against BoNT/A1. Instead of the inactivated BoNT/A1 toxoid, Halla horse was immunized with the receptor-binding domain present in the C-terminus of heavy chain of BoNT/A1 (BoNT/A1-HCR) expressed in Escherichia coli. The anti-BoNT/A1-HCR antibody titer increased rapidly by week 4, and this level was maintained for several weeks after boosting immunization. Notably, $20{\mu}l$ of the week-24 BoNT/A1-HCR(-immunized) equine serum showed an in vitro neutralizing activity of over 8 international units (IU) of a reference equine antitoxin. Furthermore, $20{\mu}l$ of equine serum and $100{\mu}g$ of purified equine $F(ab^{\prime})_2$ showed 100% neutralization of 10,000 $LD_{50}$ in vivo. The results of this study shall contribute towards optimizing antitoxin production for BoNT/A1, which is essential for emergency preparedness and response.

• Asian-Australasian Journal of Animal Sciences
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• 제32권9호
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• pp.1430-1438
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• 2019

Objective: This experiment was designed to determine the effects of coated cysteamine hydrochloride (CC) on muscle fiber characteristics, amino acid composition and transporters gene expression in the longissimus dorsi muscle (LDM) of finishing pigs. Methods: Two hundred and sixteen Duroc/Landrace/Yorkshire cross-bred male finishing pigs were fed with a corn-soybean basal diet supplemented with 0, 70, and 140 mg/kg cysteamine. Each group contained eight replicates of nine pigs per replicate. After 29 days, one pig was randomly selected from each replicate and slaughtered. Blood and LDM samples were collected and analyzed. Results: The results showed that supplemental dietary CC increased (p<0.05) the muscle fiber density. And CC supplementation also up-regulated (p<0.05) the expression of myosin heavy chain 1 (MyHC1) and MyHC2x mRNA levels, and down-regulated (p<0.05) MyHC2b expression in the LDM. Additionally, supplemental dietary CC reduced (p<0.05) the concentration of total cholesterol in the plasma and enhanced (p<0.05) the concentrations of essential amino acid and total amino acid in the LDM. The relative expression levels of chloramphenicol acetyltransferase 2, $b^{0,+}$ amino acid transporter, and $y^+$-L-type amino acid transporter 1 were upregulated (p<0.05) in the LDM when pigs were fed with the dietary CC of 70 mg/kg. Conclusion: Cysteamine supplementation could increase fiber density and distribution of fiber types. It also improved the deposition of protein in the LDM by up-regulated the expression of amino acid transporters.

• 현장농수산연구지
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• 제24권1호
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• pp.41-48
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• 2022

본 연구는 사료비의 절감을 위해 배합사료 비율을 줄이고 조사료 비율을 높이면서 한우 고급육으로 생산할 수 있는 방법을 찾아내기 위해 수행하였다. 대조구(고급육 사양)와 옥수수 사일리지 급여구(대조구 대비 배합사료 66% 급여 및 옥수수 사일리지)의 성장, 도체성적 및 등심근육 내유전자 발현에 미치는 영향에 대하여 분석했다. 옥수수 사일리지 급여는 비육 후기의 일당 증체량 및 출하체중이 수치적으로 낮게 나타났지만 유의미한 차이는 나타나지 않았다. 도체특성에서도 옥수수 사일리지 급여구가 도체중, 등심단면적 및 근내 지방도가 수치적으로 낮게 나타났지만 유의미한 차이는 나타나지 않았다. 육량 A등급 출현은 옥수수 사일리지 급여구가 23% 높게 나타났으며, 육질 1⁺⁺등급 출현율은 대조구가 17% 높게 나타났다. 이는 옥수수 사일리지 급여구가 대조구 대비 배합사료 급여량이 적음으로 인해 나타난 결과로 보여진다. 등심내 유전자 발현에서는 옥수수 사일리지 급여구가 근육발달 유전자 발현이 높게 나타났으며, 지방발달 유전자 발현에서는 대조구가 높게 나타났다. 본 연구의 결과 배합사료의 고급육 사양 대비 옥수수 사일리지 사양이 육질 1⁺⁺등급의 출현이 다소 감소하지만 육량 A등급의 출현을 높여주므로 사료비 절감의 생산 방법 중 하나로 될 수 있을 것으로 보여진다.

• 대한본초학회지
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• 제34권6호
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• pp.79-89
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• 2019

Objective : This study investigated the anti-diabetic effects of DM1, a herbal mixture with Atractylodis Rhizoma, Anemarrhenae Rhizoma, and Cinnamomi Cortex in high fat diet (HFD)-induced diabetic mice and the mechanism in C2C12 mouse skeletal muscle cells. Methods : The C57B/6 mice were fed high fat for 12 weeks, and then administrated DM1 extract (500 mg/kg, p.o.) for 4 weeks. The changes of body weight, calorie and water intakes, fasting blood glucose levels and the serum levels of glucose, insulin, triglyceride, HDL-cholesterol, AST and ALT were measured in mice. The histological changes of liver and pancreas tissues were also observed by H&E stain. C2C12 myoblasts were differentiated into myotubes and then treated with DM1 extract (0.5, 1, and 2 mg/㎖) for 24 hr. The expression of myosin heavy chain (MHC), PGC1α, Sirt1 and NRF1, and the AMPK phosphorylation were determined in the myotubes by western blot, respectively. Results : The DM1 extract administration significantly decreased the calorie and water intakes, glucose, triglyceride, AST and ALT levels and increased insulin and HDL-cholesterol in HFD-induced diabetic mice. DM1 extract inhibited lipid accumulation in liver tissue and improved glucose tolerance. In C2C12 myotubes, DM1 treatment increased the expression of MHC, PGC1α, Sirt-1, NRF-1 and the AMPK phosphorylation. Conclusion : In our results indicate that DM1 can improve diabetic symptoms by decreasing the obesity, glucose tolerance and fatty liver in HFD-induced diabetic mice, and responsible mechanism is might be related with energy enhancement.

• 현장농수산연구지
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• 제3권1호
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• pp.48-69
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• 2001

1997년 부터 한국농업전문학교에서 수집하여 재배되고 있는 마 (Dioscorea alata L.) 계통 중 품질이 우수하고 이용 가치가 기대되는 33개 계통에 대한 특성을 조사하였으며, 형태적 차이를 보이는 19개 계통에 대한 RAPD분석을 실시하였다. 1. 주당 괴경의 전체 무게는 최대 2,147g (No.36), 최소 90g (No.20)으로 평균 610g이었다. 주당 평균 괴경수는 2.8개였으며 최대 4.7개, 최소 1.3개였다. 괴경중은 평균 363g이었으며 최대 1200g, 최소 70g이었다. 2. 괴경의 육질은 백색, 담황색 및 적자색 3가지 패턴을 보였으며, 잎은 녹색, 진녹색 및 담녹색으로 분류되었다. 3. RAPD 분석에 사용된 총 113개 primers 중 12개 primer 만이 모든 분류군에서 증폭되었다. 이를 통하여 93개의 밴드를 얻었으며 69개 밴드는(71.0%) polymorphic 하게 나타났다. 4. 유집분석 결과 19개 계통은 유사도 지수 값 0.66~0.90의 범위로 나타났으며, 크게 2개의 그룹, 즉 인도네시아와 가고시마에서 도입된 8개의 계통이 포함된 그룹과, 유사도지수 0.70~0.90의 범위를 갖는 Nauru, Palau Is., Okinawa와 Papua New Guinea에서 도입된 11 계통이 포함된 그룹으로 대별되었다.

• Animal Bioscience
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• 제35권10호
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• pp.1545-1555
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• 2022

Objective: Our study aimed to investigate the effects of a 2% increase in dietary total digestible nutrients (TDN) value during the growing (7 to 12 mo of age) and fattening (13 to 30 mo of age) period of Hanwoo steers. Methods: Two hundred and twenty Hanwoo steers were assigned to one of two treatments: i) a control group (basal TDN, BTDN, n = 111 steers, growing = 70.5%, early fattening = 71.0%, late fattening = 74.0%) or high TDN (HTDN, n = 109 steers, growing = 72.6%, early = 73.1%, late = 76.2%). Growth performance, carcass traits, blood parameters, and gene expression of longissimus dorsi (LD) (7, 18, and 30 mo) were quantified. Results: Steers on the BTDN diets had increased (p≤0.02) DMI throughout the feeding trial compared to HTDN, but gain did not differ appreciably. A greater proportion of cattle in HTDN received Korean quality grade 1 (82%) or greater compared to BTDN (77%), while HTDN had a greater yield grade (29%) than BTDN (20%). Redness (a^*) of LD muscle was improved (p = 0.021) in steers fed HTDN. Feeding the HTDN diet did not alter blood parameters. Steers fed HTDN diet increased (p = 0.015) the proportion of stearic acid and tended to alter linoleic acid. Overall, saturated, unsaturated, monounsaturated, and polyunsaturated fatty acids of LD muscle were not impacted by the HTDN treatment. A treatment by age interaction was noted for mRNA expression of myosin heavy chain (MHC) IIA, IIX, and stearoyl CoA desaturase (SCD) (p≤0.026). No treatment effect was detected on gene expression from LD muscle biopsies at 7, 18, and 30 mo of age; however, an age effect was detected for all variables measured (p≤0.001). Conclusion: Our results indicated that feeding HTDN diet could improve overall quality grade while minimum effects were noted in gene expression, blood parameters, and growing performance. Cattle performance prediction in the feedlot is a critical decision-making tool for optimal planning of cattle fattening and these data provide both benchmark physiological parameters and growth performance measures for Hanwoo cattle feeding enterprises.

• 한국습지학회지
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• 제25권4호
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• pp.315-325
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• 2023

본 연구는 하천습지의 교란 과정을 근현대 지도와 항공사진을 근거로 분석하고, 왕진지구 주변 수문생태 영향권의 토지피복과 NDVI 변화를 분석하였다. 왕진나루에는 안정된 모래톱이 형성되어 제내지 농경지와 자연스럽게 연결되어 있었으나 대홍수와 제방축조, 경지정리 및 하천종합관리로 모래톱과 하천습지가 소멸된 이후 인위적인 대체습지와 생태공원이 조성되고 하중도 형태의 모래톱이 다시 생성되었다. 수문생태영향권 토지피복 변화는 농업지역의 논·밭이 2013년 36.3%에서 2022년 22.9%로 감소하였는데 면적으로는 814,476m²로 가장 많은 변화가 있는 것으로 나타났다. 시간이 경과됨에 따라 토지피복유형은 초지화가 진행되고 있는 것으로 확인되었다. 식생지수 변화를 계절별 변화를 종합해보면 전반적으로 식생지수의 변화량은 봄철(3월)이 가장 크고, 그다음은 여름철(6월)이며 수역을 제외하고는 가을철(9월)의 변화의 폭이 가장 적은 것으로 나타났다. 토지이용별로는 대체습지에서 39.1% 향상, 하중도 38.2% 향상, 생태공원 44.3% 향상, 농업지역 35.6% 향상, 수역 -8.1% 감소 등 전반적으로 식생지수(NDVI)가 증가되었다.