• Title/Summary/Keyword: Cloned pigs

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Temporal Expression of RNA Polymerase II in Porcine Oocytes and Embryos

  • Oqani, Reza;Lee, Min Gu;Tao, Lin;Jin, Dong Il
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.237-241
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    • 2012
  • Embryonic genome activation (EGA) is the first major transition that occurs after fertilization, and entails a dramatic reprogramming of gene expression that is essential for continued development. Although it has been suggested that EGA in porcine embryos starts at the four-cell stage, recent evidence indicates that EGA may commence even earlier; however, the molecular details of EGA remain incompletely understood. The RNA polymerase II of eukaryotes transcribes mRNAs and most small nuclear RNAs. The largest subunit of RNA polymerase II can become phosphorylated in the C-terminal domain. The unphosphorylated form of the RNA polymerase II largest subunit C-terminal domain (IIa) plays a role in initiation of transcription, and the phosphorylated form (IIo) is required for transcriptional elongation and mRNA splicing. In the present study, we explored the nuclear translocation, nuclear localization, and phosphorylation dynamics of the RNA polymerase II C-terminal domain in immature pig oocytes, mature oocytes, two-, four-, and eight-cell embryos, and the morula and blastocyst. To this end, we used antibodies specific for the IIa and IIo forms of RNA polymerase II to stain the proteins. Unphosphorylated RNA polymerase II stained strongly in the nuclei of germinal vesicle oocytes, whereas the phosphorylated form of the enzyme was confined to the chromatin of prophase I oocytes. After fertilization, both unphosphorylated and phosphorylated RNA polymerase II began to accumulate in the nuclei of early stage one-cell embryos, and this pattern was maintained through to the blastocyst stage. The results suggest that both porcine oocytes and early embryos are transcriptionally competent, and that transcription of embryonic genes during the first three cell cycles parallels expression of phosphorylated RNA polymerase II.

Effect of Cysteamine on In Vitro Maturation, Fertilization and Culture of Porcine Oocytes

  • Yi, Y. J;Kim, M. Y.;Lee, S. H.;T. S. Min;D. I. Jin;Park, C. S.
    • Korean Journal of Animal Reproduction
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    • v.27 no.4
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    • pp.275-280
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    • 2003
  • This study was carried out to investigate the effect of cysteamine addition during in vitro maturation, fertilization and culture of porcine oocytes. Oocytes were matured for the first 22 h in mTCM -199 media supplemented with or without 150 $\mu$M cysteamine. They then were matured for an additional 22 h in mTCM-199 media without hormones supplemented with or without 150 $\mu$M cysteamine. When cumulus-oocyte complexes (COCs) were matured in the mTCM-199 media supplemented with cysteamine, the rates of GVBD and maturation (metaphase II) were enhanced as compared to the media without the addition of cysteamine. Also, when COCs were matured in the mTCM-199 media supplemented with cysteamine, the rates of sperm penetration, male pronucleus formation, cleavage and blastocyst formation after in vitro fertilization were enhanced as compared to the media without the addition of cysteamine. In conclusion, it was suggested that oocytes matured for the first 22 h in mTCM-199 media supplemented with 150 $\mu$M cysteamine increased the rates of metaphase II, sperm penetration, male pronucleus and blastocyst formation were higher as compared to the media without addition of cysteamine.

Discrimination of Korean Cattle (Hanwoo) Using DNA Markers Derived from SNPs in Bovine Mitochondrial and SRY Genes

  • Yoon, D.;Kwon, Y.S.;Lee, K.Y.;Jung, W.Y.;Sasazaki, S.;Mannen, H.;Jeon, J.T.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.1
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    • pp.25-28
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    • 2008
  • In order to distinguish Korean cattle (Hanwoo) beef from the imported beef from Australia in Korean markets, DNA markers based on PCR-RFLP from mitochondrial genes and SRY gene were applied. A total of 2,826 beef samples comprising 1,495 Hanwoo and 1,331 foreign cattle breeds were obtained in Korea. An 801 bp fragment of the SRY gene on the bovine Y chromosome, a 343 bp fragment of ND4 gene and a 528 bp fragment of ND5 gene in the bovine mtDNA were amplified by PCR and digested with three restriction enzymes, MseI, HpyCH4III and Tsp509I, respectively. The results showed that Bos taurus (T) type was the majority in Hanwoo by combining three markers (99.5%). However, 78.2% of Bos indicus (I) type was observed in the imported beef samples. These results indicated that three markers used in this study will be used as valuable markers for discriminating imported beef against Hanwoo.

Protein Profile of Embryonic Stem Cells Following Differentiation

  • D. I. Jin;Kang, H. S;Kim, H. J.;Lee, S. H.;Park, C. S.;K. S. Im;Lee, H. M.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.377-384
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    • 2002
  • To investigate the expression patterns of proteins and growth factor signals in differentiated rabbit embryonic stem (ES) cells, ES cells with confluent stage grown of feeder layer and differentiated cells into embryoid bodies (EB) without feeder cell were applied to protein gel and Western blotting analysis. There were 66kDa and 28kDa specifically expressed in differentiated ES cell but not in undifferentiated ES cell while 25kDa protein band showed up in only undifferentiated ES cells. Also there were some difference of protein bands in several area of gel between differentiated and undifferentiated ES cells such as about 100 kDa, 50kDa and 27kDa areas, but there was no difference in band pattern of one-dimensional gel analysis between mouse ES cells and rabbit ES cells. IGF-I receptor and EGF receptor were expressed in differentiated cells and undifferentiated cells. And ICF-I and EGF were not expressed in both differentiated and undifferentiated cells. These results indicated that ES cells express their own proteins to inhibit differentiation while EB cells synthesize different proteins to differentiate, and 16F-I receptor and EGF receptor were expressed in both ES and EB cells probably for the different functions.

Comparative Study of Two Anesthetic Combinations (Zoletil/Midazolam and Zoletil/Xylazine) in Pigs (돼지에서 Zoletil/Midazolam과 Zoletil/Xylazine의 2가지 병용마취에 대한 비교 연구)

  • Jee, Hyun-Chul;Lee, Jae-Yeon;Jeong, Seong-Mok;Lee, Soo-Jin;Park, Chang-Sik;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.27 no.4
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    • pp.330-335
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    • 2010
  • This study was performed to compare the anesthetic and cardiorespiratory effects of the tiletamine/zolazepam/xylazine (TZX) combination and tiletamine/zolazepam/midazolam (TZM) combination. Eight healthy Landrace $\times$ Yorkshire pigs were randomly assigned to two groups. Each group was composed of four pigs. The pigs in group 1 (TZX) received tiletamine/zolazepam (2 mg/kg, IM) and xylazine (2 mg/kg, IM). The pigs in group 2 (TZM) received tiletamine/zolazepam (2 mg/kg, IM) and midazolam (0.5 mg/kg, IV). Induction time, anesthesia time and standing time were recorded for each pig. The scores of anesthetic effects were subjectively evaluated every 15 minutes during anesthesia. Cardiopulmonary parameters (heart rate, arterial blood pressure, respiratory rate and rectal temperature) were monitored and recorded 0, 5, 15, 30, 45 and 60 minutes after administration of drugs. Arterial blood gases ($pH_a$, $P_aCO_2$ and $P_aO_2$) and oxygen saturation ($SO_2$) were analyzed at same times. The scores of anesthetic effects decreased in the TZX group compare with the TZM group. From 5 to 85 minutes the mean heart rate in the TZX group was significantly lower than those in the TZM group. Mean arterial blood pressure in the TZX group was significantly higher than those in the TZM group at 5, 15 and 30 minutes. Both drug combinations provided a smooth induction and good immobilization. Scores of anesthetic effects in the TZM group were better than those in the TZX group. The effects to the cardiorespiratory function and temperature were lesser in the TZM group than those in the TZX group. In conclusion, when the two drug combinations were compared, the TZM group showed better anesthetic effects and less cardiorespiratory effects.

Mitochondrial DNA Diversity and Origin of Red Chittagong Cattle

  • Bhuiyan, M.S.A.;Bhuiyan, A.K.F.H.;Yoon, D.H.;Jeon, J.T.;Park, C.S.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.10
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    • pp.1478-1484
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    • 2007
  • To determine the origin and genetic diversity of Red Chittagong (RC) cattle in Bangladesh, we analyzed mitochondrial DNA displacement loop (D-loop) sequences of 48 samples along with 22 previously published sequences from Bos indicus and Bos taurus breeds. Twenty five haplotypes were identified in RC cattle that were defined by 44 polymorphic sites and nucleotide diversity was $0.0055{\pm}0.0026$. The estimated sequence divergence times between RC and other zebu cattle breeds studied ranged between 22,700-26,900 years before present (YBP) which, it is suggested, predate domestication of RC cattle. Furthermore, it is assumed that introgressions have occurred in this breed mainly from Indian zebu breeds in the recent millennia. The phylogenetic studies showed RC cattle clustered with Bos indicus lineage with two distinct haplogroups representing high genetic variability of this breed. These findings can be used for designing proper breeding and conservation strategies for RC cattle in Bangladesh.

Antagonistic Effects of Atipamezole and Yohimbine against Anesthesia with Medetomidine and Ketamine Combination in Pigs (돼지에서 Medetomidine-ketamine 마취에 대한 Atipamezole과 Yohimbine의 길항효과)

  • Lee, Jae-Yeon;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.28 no.3
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    • pp.291-296
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    • 2011
  • The aims of the present study were to investigate the anesthetic and hemodynamic effects of medetomidine-ketamine combination and to compare antagonistic effects of atipamezole and yohimbine on the recovery of pig from anesthesia induced by medetomidine-ketamine combination. Landrace and Yorkshire cross-bred pigs were evaluated in the present study. Pigs (n = 8) received three different treatments (one treatment per 14 days in a random order). All pigs were injected intramuscularly with medetomidine, and ketamine in a single syringe. Intravenous injections of atipamezole (MKA), yohimbine (MKY), or a control saline solution (MK) were administered 20 minutes after the medetomidine-ketamine combination injection. The intravenous antagonist injections quickly reversed the medetomidine-ketamine induced sedation in the pigs, resulting in a significantly shorter duration of anesthesia in the MKA and MKY groups compared to the MK group. Mean arterial pressure (MAP) levels were significantly lower in the MKA and MKY groups compared to the MK group. Scores for posture and responses to noxious stimuli after atipamezole and yohimbine administration were significantly lower in the MKA and MKY groups than in the MK. In conclusion, the sedative effects and increases in blood pressure induced by a medetomidine-ketamine combination were quickly and smoothly reversed by atipamezole or yohimbine.