• Title/Summary/Keyword: Cloned pigs

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Identification of Differentially Expressed Genes Related to Intramuscular Fat Development in the Early and Late Fattening Stages of Hanwoo Steers

  • Lee, Seung-Hwan;Park, Eung-Woo;Cho, Yong-Min;Kim, Sung-Kon;Lee, Jun-Heon;Jeon, Jin-Tae;Lee, Chang-Soo;Im, Seok-Ki;Oh, Sung-Jong;Thompson, J.M.;Yoon, Du-Hak
    • BMB Reports
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    • v.40 no.5
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    • pp.757-764
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    • 2007
  • Marbling of cattle meat is dependent on the coordinated expression of multiple genes. Cattle dramatically increase their intramuscular fat content in the longissimus dorsi muscle between 12 and 27 months of age. We used the annealing control primer (ACP)-differential display RT-PCR method to identify differentially expressed genes (DEGs) that may participate in the development of intramuscular fat between early (12 months old) and late fattening stages (27 months old). Using 20 arbitrary ACP primers, we identified and sequenced 14 DEGs. BLAST searches revealed that expression of the MDH, PI4-K, ferritin, ICER, NID-2, WDNMI, telethonin, filamin, and desmin (DES) genes increased while that of GAPD, COP VII, ACTA1, CamK II, and nebulin decreased during the late fattening stage. The results of functional categorization using the Gene Ontology database for 14 known genes indicated that MDH, GAPD, and COP VII are involved in metabolic pathways such as glycolysis and the TCA cycle, whereas telethonin, filamin, nebulin, desmin, and ACTA1 contribute to the muscle contractile apparatus, and PI4-K, CamK II, and ICER have roles in signal transduction pathways regulated by growth factor or hormones. The final three genes, NID-2, WDNMI, and ferritin, are involved in iron transport and extracellular protein inhibition. The expression patterns were confirmed for seven genes (MDH, PI4-K, ferritin, ICER, nebulin, WDNMI, and telethonin) using real-time PCR. We found that the novel transcription repressor ICER gene was highly expressed in the late fattening stage and during bovine preadipocyte differentiation. This information may be helpful in selecting candidate genes that participate in intramuscular fat development in cattle.

Effect of Electroacupuncture Analgesia on Changes of Vital Signs and Blood Chemical Values in Cats

  • Shin, Dong-Hoon;Lee, Jae-Yeon;Kim, Duck-Hwan;Park, Chang-Sik;Jeong, Seong-Mok;Son, Dong-Soo;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.25 no.3
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    • pp.170-175
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    • 2008
  • The present study was performed to investigate the anesthetic or analgesic effect of tiletamine-zolazepam (TZ) and electro acupuncture analgesia (EAA) in cats. Twelve healthy cats were randomly assigned to receive either TZ or EA. TZ group cats with weight of $3.65{\pm}0.48kg$ received 10.0 mg/kg of TZ intramuscularly. EA group cats with weight of $3.62{\pm}0.52kg$ received 5V, 30Hz and 60 minutes of EA. The acupoints used were Tian-ping (GV-5, +), Bai-hui (GV-20, -). Therefore, after and before experiment, some serum chemistry profiles (alkaline phospatase, aspartate aminotransferase, alanine aminotransferase, glucose and total protein) and change of vital signs (rectal temperature, heart rate, respiratory rate) were examined. All cats were examined pre, and 5, 25, 65 and 105 minutes after administration of TZ or operation of EA. The cats in EA group showed a smaller change in rectal temperature, heart rate and respiratory rate than in the TZ group (p<0.05). In both groups, total protein concentration was constant throughout the period of anesthesia, and the serum glucose increased gradually throughout the period of anesthesia. However, the cats in EA group showed a smaller change in alkaline phospatase, aspartate aminotransferase and alanine aminotransferase within the limit of safety than in the TZ group (p<0.05). While coming to induction, the TZ group took a mean $2.4{\pm}0.7$ minutes to achieve sternal recumbency, compared with $10.5{\pm}2.1$ minutes by the EA group, and $3.2{\pm}0.6$ minutes to achieve lateral recumbency, compared with $18.8{\pm}1.9$ minutes by the EA group (p<0.05). When recovering from anesthesia, the TZ group took $164.3{\pm}17.9$ minutes to achieve sternal position time, compared with $67.7{\pm}4.6$ minutes by the EA group, and $202.0{\pm}15.7$ minutes to stand, compared with $73.0{\pm}6.1$ minutes for the EA group (p<0.05). In this study, the cats anesthetized with EA showed a more rapid recovery rather than the cats under TZ anesthesia. Also, there do not appear to be any negative physiologic effects associated with acupuncture-induced surgical analgesia. So, it was considered that EAA may be used effectively in shock, debilitated cats, as compared to TZ.

The Porcine FoxO1, FoxO3a and FoxO4 Genes: Cloning, Mapping, Expression and Association Analysis with Meat Production Traits

  • Yu, Jing;Zhou, Quan-Yong;Zhu, Meng-Jin;Li, Chang-Chun;Liu, Bang;Fan, Bin;Zhao, Shu-Hong
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.627-632
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    • 2007
  • FoxO1, FoxO3a and FoxO4 belong to the FoxO gene family, which play important roles in the PI3K/PKB pathway. In this study, we cloned the porcine FoxO1, FoxO3a and FoxO4 sequences and assigned them to SSC11p11-15, SSC1p13 and SSC xq13 using somatic cell hybrid panel (SCHP) and radiation hybrid panel (IMpRH). RT-PCR results showed that these three genes are expressed in multiple tissues. Sequencing of PCR products from different breeds identified a synonymous T/C polymorphism in exon 2 of FoxO3a. This FoxO3a single nucleotide polymorphism (SNP) can be detected by AvaII restriction enzyme. The allele frequencies of this SNP were investigated in Dahuabai, Meishan, Tongcheng, Yushan, Large White, and Duroc pigs. Association of the genotypes with growth and carcass traits showed that different genotypes of FoxO3a were associated with carcass length and backfat thickness between 6th and 7th ribs (BTR) and drip loss (p<0.05).

Effects of Castration on Androgen Receptor, IGF-I Ea, MGF and Myostatin Gene Expression in Skeletal Muscles of Male Pigs

  • Yao, Yuchang;Cai, Zhaowei;Zhang, Lifan;Zhao, Chunjiang;Wu, Keliang;Xu, Ningying;Liu, Gang;Wu, Changxin
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.8
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    • pp.1069-1077
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    • 2009
  • Castration of male pig produces significant negative effects on skeletal muscle development. The androgen receptor (AR), two splice variants of insulin-like growth factor-I (IGF-I Ea and MGF) and the myostatin gene may play important roles in this process. In the present study, the expression of AR, IGF-I Ea, MGF and myostatin genes in three skeletal muscles, the brachialis, longissimus and semitendinosus, were studied using real-time quantitative RT-PCR. Our experimental design used 14 pairs of male Landrace sire${\times}$Yorkshire dam piglets. The two piglets in each pair were full sibs, one of which was castrated at 21 d of age; the other remained intact. The study group was divided into subgroups of equal size. Animals in the first subgroup were slaughtered at 147 d and those of the second at 210 d of age. Carcass weight and lean meat yield were similar between boars and barrows at 147 d of age (p>0.05), whereas barrows had lower carcass weight and less lean meat yield at 210 d of age (p<0.05). Castration caused down-regulation of AR gene expression at both 147 and 210 d of age (p<0.05). The two splice variants of the IGF-I gene from porcine skeletal muscle were cloned using RT-PCR, and it was found that MGF differs from IGF-I Ea in having a 52-base insert in the last coding exon of the mRNA. Both splice variants were down-regulated by castration only at 210 d of age (p<0.05). No differences in expression of the myostatin gene were observed between boars and barrows at either 147 or 210 d of age (p>0.05). These results suggest that the downregulation of AR, IGF-I Ea and MGF gene expression following castration helps to explain the negative effect of castration on skeletal muscle development.

Effects of Corn Distiller's Dried Grains with Solubles on Production and Egg Quality in Laying Hens

  • Cheon, Y.J.;Lee, H.L.;Shin, M.H.;Jang, A.;Lee, S.K.;Lee, J.H.;Lee, B.D.;Son, C.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.9
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    • pp.1318-1323
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    • 2008
  • Corn distiller's dried grains with solubles (DDGS) is a completely new feed ingredient in the Korean feed market. There is an ever increasing need for the Korean feed industry to import and make the best of it as a high protein and high energy feed ingredient. A layer feeding trial was conducted for 10 weeks to investigate the effects of addition of light-colored DDGS to layer diets on laying performance, egg qualities and yolk fatty acid composition. Also, the economics of using DDGS in the Korean situation was analyzed. Nine hundred Hy-line Brown layers, 24 weeks of age, were employed in a feeding trial consisting of four dietary treatments (0, 10, 15, and 20% DDGS), and five replicates per treatment. All experimental diets were prepared as iso-protein (17%) and iso-calorie (TMEn 2,780 kcal/kg). The use of DDGS up to 20% in layer diets did not exert any influence on feed intake, laying rate, total egg mass, mean egg weight and feed conversion ratio (p>0.05). The color and breaking strength of eggshell, as well as the albumin height and Haugh unit were not affected by the addition of DDGS up to 20% in the diet. The yolk color was significantly increased by DDGS supplementation (p<0.05). As the DDGS level increased, the oleic acid content decreased, and the linoleic acid increased (p<0.05) in egg yolk. The degree of saturation of yolk fatty acids was not affected by DDGS supplementation. The inclusion of light-colored DDGS up to 20% in layer diets resulted in a decrease of feed cost per kg without any undesirable effect on laying performance. In conclusion, the light-colored DDGS (L* 56.65) could be used up to 20% in layer diets without any harmful effect on laying performance, and possibly provide economic benefits to the Korean poultry industry.

Novel splice isoforms of pig myoneurin and their diverse mRNA expression patterns

  • Guo, Xiaohong;Li, Meng;Gao, Pengfei;Cao, Guoqing;Cheng, Zhimin;Zhang, Wanfeng;Liu, Jianfeng;Liu, Xiaojun;Li, Bugao
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.10
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    • pp.1581-1590
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    • 2018
  • Objective: The aim of this study was to clone alternative splicing isoforms of pig myoneurin (MYNN), predict the structure and function of coding protein, and study temporal and spatial expression characteristics of each transcript. Methods: Alternative splice isoforms of MYNN were identified using RNA sequencing (RNA-seq) and cloning techniques. Quantitative real-time polymerase chain reaction (qPCR) was employed to detect expression patterns in 11 tissues of Large White (LW) and Mashen (MS) pigs, and to study developmental expression patterns in cerebellum (CE), stomach (ST), and longissimus dorsi (LD). Results: The results showed that MYNN had two alternatively spliced isoforms, MYNN-1 (GenBank accession number: KY470829) and MYNN-2 (GenBank accession number: KY670835). MYNN-1 coding sequence (CDS) is composed of 1,830 bp encoding 609 AA, whereas MYNN-2 CDS is composed of 1,746 bp encoding 581 AA. MYNN-2 was 84 bp less than MYNN-1 and lacked the sixth exon. MYNN-2 was found to have one $C_2H_2$ type zinc finger protein domain less than MYNN-1. Two variants were ubiquitously expressed in all pig tissues, and there were significant differences in expression of different tissues (p<0.05; p<0.01). The expression of MYNN-1 was significantly higher than that of MYNN-2 in almost tissues (p<0.05; p<0.01), which testified that MYNN-1 is the main variant. The expression of two isoforms decreased gradually with increase of age in ST and CE of MS pig, whereas increased gradually in LW pig. In LD, the expression of two isoforms increased first and then decreased with increase of age in MS pig, and decreased gradually in LW pig. Conclusion: Two transcripts of pig MYNN were successfully cloned and MYNN-1 was main variant. MYNN was highly expressed in ST, CE, and LD, and their expression was regular. We speculated that MYNN plays important roles in digestion/absorption and skeletal muscle growth, whereas the specific mechanisms require further elucidation.

The Search of Pig Pheromonal Odorants for Biostimulation Control System Technologies: Prediction of Pig Pheromonal Tetrahydrofuran-2-yl Family Compounds by Means of Ligand Based Approach (생물학적 자극 통제 수단으로 활용하기 위한 돼지 페로몬성 냄새 물질의 탐색: Ligand Based Approach에 의한 돼지 페로몬성 Tetrahydrofuran-2-yl 계 화합물의 예측)

  • Soung, Min-Gyu;Cho, Yun-Gi;Park, Chang-Sik;Sung, Nack-Do
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.141-146
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    • 2008
  • To search a new porcine pheromonal odorant, the models of four type (2D-QSAR, HQSAR, CoMFA & CoMSlA) were derived from quantitative structure-activity relationship (QSAR) between tetrahydrofuran-2-yl family compounds and their observed binding affinity constants (Obs.p$[Od]_{50}$). The optimized CoMFA model (predictability; $r^{2}_{cv.}(q^2)$=0.886 & correlation coefficient: $r^{2}_{ncv.}$=0.984) from ligand based approaches was confirmed as the best model among them. The $N^{1}$-allyl-$N^{2}$-(tetrahydrofuran-2-yl)methyl)oxalamide (P1), 2-(4-trimethylammoniummethylcyclohexyloxy)tetrahydrofurane (P5) and 2-(3-trimethylammoniummethylcyclohexyloxy)tetrahydrofurane (P6) molecules predicted as porcine pheromonal odorant by the CoMFA model were showed relatively high binding affinity constant values (Pred.p$[Od]_{50}=8{\sim}10$) and very lower toxicity values against some sorts of toxicity.

Establishment and Maintenance of Embryonic Stem-like Cell Lines from In Vitro Produced Bovine Blastocysts (체외수정 유래 소 배반포로부터 유사 배아 줄기 세포의 확립 및 유지)

  • Lee, Yu-Yeon;Kim, Sun-Uk;Kim, Ji-Su;Song, Bong-Seok;Cho, Yoon-Jeong;Park, Jung-Sun;Yu, Dae-Yeul;Jin, Dong-Il;Lee, Kyung-Kwang;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.215-220
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    • 2007
  • This study was conducted to examine the establishment of bovine ES-like cells having pluripotency. The hatched blastocysts derived from culture of in vitro fertilized embryos for 10 to 12 days dissociated mechanically into ICM-and trophectoderm-rich clumps using needle, and cultured onto mitotically-inactivated MEF feeder layer. The primary colonies originated from ICM cells were detached mechanically 7 days after seeding and subsequent subculture was conducted at intervals of every 5 to 7 days. Two ES -like cell lines were established and maintained over 40 passages. Self-renewal of the established lines was confirmed by examining the alkaline phosphatase activity, stem cell-specific marker profiles including SSEA isotopes, Oct-4 and STAT3. Moreover, the established cell lines could produce anchorage-independent embryoid bodies (EBs) with gradual decrease of Oct-4 transcript level in time-dependent manner.

Immunohistochemical Study on the TNFα-Secreting Macrophages and Endothelial Cells in the Porcine Corpus Luteum (돼지 황체에서 TNFα-분비 대식세포와 내피세포에 관한 면역조직화학적 연구)

  • Park, C.S.;Han, S.R.;Kim, S.I.;Cho, K.J.;Kim, W.S.
    • Journal of Animal Science and Technology
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    • v.47 no.5
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    • pp.711-720
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    • 2005
  • In the corpus luteum, TNF$\alpha$ is known to induce functional and structural luteolysis. In addition, it acts as luteotropic agent during the initial and early stage of luteal development. In spite of its importance in corpus luteal development, there is still different opinions for the source cells of TNF$\alpha$ in the corpus luteum. One is the macrophages only, and the other is macrophages are the main source and endothelial cells are the minor source. In this experiment, using the porcine corpora lutea of pregnancy and ovulatory stages, hematoxylin-eosin stain, macrophage and TNF$\alpha$ immunohistochemistry were carried to reveal the sources of TNF$\alpha$. As a result, MAC 387-positive macrophages were present in all the stages of corpora lutea. In the mature corpora lutea of nonpregnant stages, the sites of MAC 387-positive macrophages and those of TNF$\alpha$- positive macrophages were coincided, and the sites of endothelial cells and those of TNF$\alpha$-positive endothelial cells were nearly coincided. But, in the mature CL of pregnant stage, mid- and advanced luteolytic stages of both nonpregnant and pregnant stages, the sites of MAC 387-positive macrophages and those of TNF$\alpha$-positive macrophages were coincided, but not in the endothelial cells. Accordingly, it can be concluded that macrophages are the main source of TNF$\alpha$ in the corpus luteum and endothelial cells are the minor source in the mature and mid-lytic stages, but, in the advanced luteolytic stage, macrophages are the only source of TNF$\alpha$.

The Search of Pig Pheromonal Odorants for Biostimulation Control System Technologies: III. Comparative Molecular Field Analysis (CoMFA) on Binding Affinities between Ligands of 2-(Cyclohexyloxy) Tetrahydrofurane Derivatives and Porcine Odorant Binding Protein (생물학적 자극 통제 수단으로 활용하기 위한 돼지 페로몬성 냄새 물질의 탐색: III. 2-(Cyclohexyloxy) Tetrahydrofurane 유도체와 Porcine Odorant Binding Protein 사이의 결합 친화력에 관한 비교 분자장 분석)

  • Sung Nack-Do;Park Chang-Sik;Jung Hoon-Sung;Seong Min-Kyu
    • Reproductive and Developmental Biology
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    • v.30 no.1
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    • pp.13-19
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    • 2006
  • To search of new porcine pheromonal odorants for biostimulation control system technologies to improve reproductive efficiency in livestock species, the comparative molecular field analysis (CoMFA) for binding affinity constant $(p(Od)_{50})$ between porcine odorant binding protein (pOBP) and ligands of odorant 2-(cyclohexyloxy) tetrahydrofurane derivatives as substrate molecule was conducted and discussed. In the optimized CoMFA model AIV with chirality $(C_1'(R),\;C_2(S))$ in substrate molecule and atom based fit alignment (A) of odorants, the statistical results showed the best predictability of the binding affinities $(p(Od)_{50})$ based on the LOO cross-validated value $r^2_{cv}.\;(q^2=0.886)$ and non-cross-validated conventional coefficient $(r^2_{ncv}.=0.984)$. the binding affinity constants exhibited a good correlation with steric (40.8%), electrostatic (14.6%) and hydrophobic (44.6%) factors of the substrate molecules. from the analytical results of the contour maps, which may give us some valuable informations to the modification of odorants for effective binding affinity.