• Toxicological Research
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• 제24권1호
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• pp.59-68
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• 2008

N-ethyl-N-nitrosoures (ENU) is effective in inducing hypermorphic mutation as well as hypomorphic and antimorphic mutations. Therefore, this mutagen is used to the production of mutant in the mice. In order to perform an effective ENU mutagenesis using BALB/cAnN mice, determination of optimal dosage and dosage regimen of ENU is necessary. And this study tried to develop a suitable screening method and searched for novel and various mutants as model animals in phenotypedriven ENU mutagenesis. We have carried out dosage regimen for mutagenizing dose of 200 mg/kg ENU in the BALB/c mice. Total screened mice were 30,133. As the results of Esaki and Cho's Phenotype Screening, we got 2,516 phenotypic and behavior abnormalities in $G_1,\;G_2\;and\;G_3$ mice. One hundred thirty five $G_1$ phenodeviants were tested for inheritance and 16 dominant mutants were discovered. Forty two recessive mutants were also found in tested 201 micropedigrees. Early-onset mutant mice included the dysmorphology of face, eye, tail, limb, skin, and foot and abnormal behavior like circling, swimming, head tossing, stiff-walking, high cholesterol level, and tremor etc. In this study we could effectively screen $G_3$ recessive mutants. The frequent and concise early-onset screening before weaning will be available for ENU mutagenesis.

• Journal of Microbiology and Biotechnology
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• 제33권12호
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• pp.1648-1656
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• 2023

We have previously observed that feeding with single-cell hemoprotein (heme-SCP) in dogs (1 g/day for 6 days) and broiler chickens (1 ppm for 32 days) increased the proportion of lactic acid bacteria in the gut while reducing their body weights by approximately 1~2%. To define the roles of heme-SCP in modulating body weight and gut microbiota, obese C57BL/6N mice were administered varied heme-SCP concentrations (0, 0.05, and 0.5% heme-SCP in high fat diet) for 28 days. The heme-SCP diet seemed to restrain weight gain till day 14, but the mice gained weight again later, showing no significant differences in weight. However, the heme-SCP-fed mice had stiffer and oilier bodies compared with those of the control mice, which had flabby bodies and dull coats. When mice were dissected at day 10, the obese mice fed with heme-SCP exhibited a reduction in subcutaneous fat with an increase in muscle mass. The effect of heme-SCP on the obesity-associated dyslipidemia tended to be corroborated by the blood parameters (triglyceride, total cholesterol, and C-reactive protein) at day 10, though the correlation was not clear at day 28. Notably, the heme-SCP diet altered gut microbiota, leading to the proliferation of known anti-obesity biomarkers such as Akkermansia, Alistipes, Oscillibacter, Ruminococcus, Roseburia, and Faecalibacterium. This study suggests the potential of heme-SCP as an anti-obesity supplement, which modulates serum biochemistry and gut microbiota in high-fat diet-induced obese mice.

• Asian-Australasian Journal of Animal Sciences
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• 제28권8호
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• pp.1084-1089
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• 2015

Chemerin, highly expressed in adipose and liver tissues, regulates glucose and lipid metabolism and immunity in these tissues in ruminants and mice. Our previous reports showed that chemerin is involved in adipogenesis and lipid metabolism in adipose tissue as an adipokine. The aim of the present study was to identify single nucleotide polymorphisms (SNPs) in the coding region of the chemerin gene and to analyze their effects on carcass traits and intramuscular fatty acid compositions in Japanese Black cattle. The SNPs in the bovine chemerin gene were detected in 232 Japanese Black steers (n = 161) and heifers (n = 71) using DNA sequencing. The results revealed five novel silent mutations: NM_001046020: c.12A>G (4aa), c.165GT (92aa), c.321 A>G (107aa), and c.396C>T (132aa). There was no association between 4 of the SNPs (c.12A>G [4aa], c.165GG [107aa], and c.396C>T) and carcass traits or intramuscular fatty acid compositions. Regarding the remaining SNP, c.276C>T, we found that cattle with genotype CC had a higher beef marbling score than that of cattle with genotype CT, whereas cattle with genotype CT had a higher body condition score (p<0.10). Further, cattle with genotype CC had significantly higher C18:0 content in their intramuscular fat tissue than that of cattle with genotype CT (p<0.05). On the other hand, cattle with genotype CT had significantly higher C14:0 and C16:0 content in their intramuscular fat tissue (p<0.05). Moreover, the number of individuals carrying the minor allele of c.276C>T SNP is small. It is suggested that the c.276C>T SNP of the chemerin gene has potential in cattle breeding using modern methods, such as marker assisted selection. So, further functional and physiological research elucidating the impact of the chemerin gene on bovine lipid metabolism including fatty acid synthesis will help in understanding these results.

• Nutrition Research and Practice
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• 제13권4호
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• pp.286-294
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• 2019

BACKGROUND/OBJECTIVES: Docosahexaenoic acid (DHA), an n-3 long chain polyunsaturated fatty acid (LCPUFA), is acquired by dietary intake or the in vivo conversion of ${\alpha}$-linolenic acid. Many enzymes participating in LCPUFA synthesis are regulated by peroxisome proliferator-activated receptor alpha ($PPAR{\alpha}$). Therefore, it was hypothesized that the tissue accretion of endogenously synthesized DHA could be modified by $PPAR{\alpha}$. MATERIALS/METHODS: The tissue DHA concentrations and mRNA levels of genes participating in DHA biosynthesis were compared among $PPAR{\alpha}$ homozygous (KO), heterozygous (HZ), and wild type (WT) mice (Exp I), and between WT mice treated with clofibrate ($PPAR{\alpha}$ agonist) or those not treated (Exp II). In ExpII, the expression levels of the proteins associated with DHA function in the brain cortex and retina were also measured. An n3-PUFA depleted/replenished regimen was applied to mitigate the confounding effects of maternal DHA. RESULTS: $PPAR{\alpha}$ ablation reduced the hepatic Acox, Fads1, and Fads2 mRNA levels, as well as the DHA concentration in the liver, but not in the brain cortex. In contrast, $PPAR{\alpha}$ activation increased hepatic Acox, Fads1, Fads2, and Elovl5 mRNA levels, but reduced the DHA concentrations in the liver, retina, and phospholipid of brain cortex, and decreased mRNA and protein levels of the brain-derived neurotrophic factor in brain cortex. CONCLUSIONS: LCPUFA enzyme expression was altered by $PPAR{\alpha}$. Either $PPAR{\alpha}$ deficiency or activation-decreased tissue DHA concentration is a stimulus for further studies to determine the functional significance.

• 생명과학회지
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• 제29권10호
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• pp.1062-1070
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• 2019

마늘(Allium sativum)의 주요 생리활성 성분들은 다양한 종류의 암에 대해 항암효과가 보고되고 있다. 본 연구에서는 활성추적분리방법(activity-guided purification)을 이용하여 마늘의 항암성분을 발굴하고자 하였다. 마늘 에탄올 추출물을 칼럼크로마토그래피로 얻은 각각의 분획물에 대해서 AGS세포의 증식 억제율을 검증하여 가장 효과가 좋은 분획물로부터 물질을 순수분리하여 구조를 동정한 결과 N-benzyl-N-methyldecan-1-amine (NBNMA)로 밝혀졌다. NBNMA의 암생장 억제효능을 검증하기 위해서 CT-26, AGS, HepG2, HCT-116, MCF7, B16F10 및 Sarcoma-180 세포에 대한 in vitro 효과와 CT-26 결장암 세포를 마우스에 이식한 다음 in vivo 효과를 조사하였다. NBNMA는 Bcl-2의 down-regulation과 Bad의 up-regulation을 유도하여 CT-26 세포의 세포사멸 촉진시켰다. 또한, NBNMA는 세포사멸의 외적 및 내적 경로에서 caspases 억제자인 caspase 3과 caspase 9의 활성을 약간 증가시켰다. CT-26세포를 이식한 쥐에 $19.13{\mu}M/kg$의 NBNMA를 21일 동안 경구투여한 결과 암종의 크기가 43% 감소하였다. NBNMA는 in vitro 및 in vivo에서 항암 효과를 나타내었는데, 이러한 결과는 마늘로부터 순수분리한 NBNMA가 대장암치료를 위한 항암제 후보물질로서 활용 가능성이 있을 것으로 기대된다.

• 생명과학회지
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• 제33권4호
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• pp.315-324
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• 2023

폴리코사놀의 콜레스테롤 감소 효과에 대한 기전은 아직 명확하게 규명되지 못하고 있다. 최근 몇몇 연구들은 지방산 합성 경로를 통한 콜레스테롤 합성의 조절에 sterol regulatory element-binding proteins(SREBP-1c)의 역할을 제시하였다. 그러나 현재까지 SREBP-1c에 의해 조절되는 지방산 합성에서 폴리코사놀의 효과에 대한 연구는 전무하다. 그러므로 본 연구의 목표는 SREBP-1c에 의해 매개되는 지방산 합성이 폴리코사놀의 콜레스테롤 감소 효과와 관계하는지를 조사하는 것이다. 7주령의 C57BL/6 수컷 생쥐를 7개군(군당 7마리)으로 나누고 8주간 다음과 같이 처리하였다; 1) 정상식이군(정상 대조군), 2) 고지방식이군(high-fat diet, HFD, 음성대조군), 3) 고지방식이+에탄올처리군(Pol-0), 4) 고지방식이+1 mg/kg 폴리코사놀처리군(Pol-1), 5) 고지방식이+2 mg/kg 폴리코사놀처리군(Pol-2), 6) 고지방식이+4 mg/kg 폴리코사놀처리군(Pol-4),7) 고지방식이+simvastatin 50 ㎍/kg 처리군(양성 대조군). 폴리코사놀과 simvastatin은 고지방식이를 유지하는 동안 매일 동일 시간에 처리하였으며 체중과 음식섭취량은 8주 동안 매주 측정하였다. 8주 후, 혈중 콜레스테롤 수치를 측정하였으며, 간의 조직학적 분석과 SREBP-1c와 지방산의 발현을 조사하였다. 폴리코사놀은 농도-의존적으로 체중과 음식섭취량을 감소시켰다. 혈중 콜레스테롤 수치는 Pol-1과 Pol-4군에서 유의하게 감소하였으며, SREBP-1c와 FAS의 발현 역시 Pol-4 군에서 유의하게 감소하였다. 이러한 결과들은 폴리코사놀의 콜레스테롤 감소 효과가 SREBP-1c와 FAS의 발현 억제에 기인하여 일어날 수 있음을 시사한다.

• Biomolecules & Therapeutics
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• 제3권4호
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• pp.266-272
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• 1995

The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently through the use of specific antibodies. To generate and characterize a moloclonal antibody against $\beta$-adrenergic receptor, a synthetic $\beta$2-adrenergic receptor peptide (Phe-Gly-Asn-Phe-Trp-Cys-Phe-Trp-Thr-Ser-lle-Asp-Val-Leu) which may comprise part of $\beta$-adrenergic receptor ligand binding pocket was coupled to Keyhole Limpet Hemocyanin (KLH) and used as an immunogen. Male BALB/C mice were immunized with this antigen and the immunized spleen was fused with myeloma SP2/0-Ag14 cells to produce monoclonal antibodies. Two clones were obtained but one of monoclonal antibodies, mAb5G09, was used throughout in this study because the other clone, mAb5All showed weak immunoreactivity against KLH as well. The mouse monoclonal antibody mAb5G09 produced in this study showed immunoreactivity to peptide-KLH conjugates and also to human A43l cells and guinea pig lung $\beta$2-adrenergic receptor as revealed by ELISA and western blot. In the course of determination of the effects of mAb5G09 on $\beta$-receptor ligand binding, it was observed that mAb5G09 specifically bound $\beta$-adrenergic radioligand [$^3$H]dihydroalprenolol (DHA) with a dissociation constant (Kd) of 60 nM. The [$^3$H]DHA binding activity of mAb5G09 had characteristics of immunoglobulins and the binding activity was not observed in the control anti-KLH monoclonal antibody. The monoclonal antibody, mAb5G09 produced in this study may provide useful models for the study of the structure of receptor binding sites.