• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.33-33
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• 2003

This study was conduct to compare the efficacy to produce male and female somatic cloned piglets. Maturation of porcine COCs was accomplished by incubation in NCSU-23 medium supplemented with 0.6 mM cysteine, 10% porcine follicular fluid, 1mM dibutyryl cyclic adenosine monophosphate (dbc-AMP, Sigma, USA), and 0.1 IU/ml human menopausal gonadotrophin (hMG, Teikokuzoki, Japan) for 20h and then cultured without dbcAMP and hMG for another 18 to 24 h. Female and male fetal cells were isolated from each fetus, cultured in ES-DMEM medium containing 10% FCS. Enucleated oocytes were fused with fetal fibroblasts (passage 4 to 15). Reconstructed embryos were cultured in NCSU-23 with 4 mg/ml BSA under mineral oil at 39$^{\circ}C$ in 5% $CO_2$ in air. A total of 12,328 nuclear-transferred embryos (1- to 4-cell stage) were surgically transferred into 69 surrogate gilts. Three recipients aborted during the period of conception. Three gilts delivered eleven female piglets, and five recipients gave rise to birth 22 male piglets. The average birth weigh of the cloned piglets was 1.52 kg (1.38~1.83 kg) in female piglets and 0.84 kg (0.45~1.25 kg) in male piglets. Alive cloned pigs was seven in female piglets (63.6%) and four in male piglets (18.2%). The other two recipients is ongoing. This study suggests that female-derived fetal cell as a nuclear donor has more capability on production of cloned piglets than male.

• Asian-Australasian Journal of Animal Sciences
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• 제19권5호
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• pp.632-637
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• 2006

Oviduct-specific glycoprotein 1 (OVGP1) is implicated in playing a role in fertilization and early embryo development. In this study, we have obtained the sequence of intron 9 of OVGP1 gene in swine. Comparative sequencing of Meishan (a native Chinese breed) and Large White pig breeds revealed an A/T substitution at position 943. A PCR-EcoRI-RFLP assay was developed to detect this mutation. Polymorphism analysis in Qingping animals showed that pigs with BB genotype had lower number of piglets born alive (NBA) in multiple parities than pigs with AA (p<0.05) and AB genotype (p<0.01). In Large $White{\times}Meishan$ ($LW{\times}M$) $F_2$ offspring, the weight of both ovaries (OW) of the BB genotype was significantly lighter than that of AB (p = 0.05) and AA (p<0.01) genotypes. Analysis of the data also revealed that the mutation locus affected these two traits mostly by additive effects. These studies indicated that the polymorphism was associated with NBA and OW in two distinct populations and further investigations in more purebreds or crossbreds are needed to confirm these results.

• Asian-Australasian Journal of Animal Sciences
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• 제22권11호
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• pp.1495-1503
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• 2009

This experiment was conducted to determine the effect of dietary supplementation with BCAA (branched-chain amino acids: leucine, isoleucine and valine) on improving the growth of rats in a malnutritional IUGR (Intrauterine Growth Retardation) model, which was established by feeding restriction. In the experimental treatment, rats were fed purified diets supplemented with BCAA (mixed) during the whole gestation period, while arginine and alanine supplementation were set as the positive and negative control group, respectively. The results showed that, compared to the effect of alanine, BCAA reversed IUGR by increasing the fetus weights by 18.4% and placental weights by 18.0% while fetal numbers were statistically increased. Analysis of gene and protein expression revealed that BCAA treatment increased embryonic liver IGF-I expression; the uterus expressed higher levels of estrogen receptor-$\alpha$ (ER-$\alpha$) and progesterone receptor (PR), and the placenta expressed higher levels of IGF-II. Amino acid analysis of dam plasma revealed that BCAA supplementation effectively enhanced the plasma BCAA levels caused by the feed restriction. BCAA also enhanced the embryonic liver gluconeogenesis by augmenting the expression of two key enzymes, namely fructose-1,6-biphosphatase (FBP) and phosphoenolpyruvate carboxykinase (PEPCK). In conclusion, supplementation of BCAA increased litter size, embryonic weight and litter embryonic weight by improving the dam uterus and placental functions as well as increasing gluconeogenesis in the embryonic liver, which further provided energy to enhance the embryonic growth.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.73-73
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• 2001

This study was to examine whether the vitrified, one-step diluted and direct transferred Hanwoo IVM/IVF/IVC blastocysts can be successfully survived in vivo and they were succeeded into the live birth. For vitrification, blastocysts were serially exposed in glycerol (G) or/and ethylene glycol (EG) mixtures [10% (v/v) G for 5 min, 10% G plus 20% EG (v/v) for 5 min, and 25% G plus 25% EG (v/v) for 30 sect] which is diluted in 10% FBS added D-PBS. Thawing of straw was carried out in air for 10 sec and then in water bath of $25^{\circ}C$ for 20 sec. One-step dilution within the straw was done in water bath of $25^{\circ}C$ for 1 min. Vitrified and one-step diluted embryos were directly transferred into 36 (natural or hormone induced synchronized) recipient cows in 6 areas of Kyungsang Buk-Do. Pregnancies were confirmed at first when recipient cows did not return to the subsequent estrus cycle, and later by manual palpation per rectum on day 45, 90 and then living calves were derived into parturition. Overall pregnancy was 33.3%(12/36), However, higher pregnancy was obtained when the recipients exhibited estrus one day earlier than the age of transferred embryos (53.3 vs 25.0-27.3%), irrespective of synchronization methods. Also, parous recipients became pregnant higher than nulliparous heifers, And, there were not different in pregnancy rates by the aspect of corpus luteum (CL) quality of recipients (good, 29.4; fair, 37.5; poor, 33.3%). One hundred eight of frozen-thawed Hanwoo blastocysts were directly transferred into 36 recipient cows. In 12 of pregnant cows, 3 cows were aborted and 9 cows were calved [single, 66.7% (6/9): twin, 33.3% (3/9)]. Total embryo implantation rate was 11.1% (12/108). However, 9 Hanwoo calves were lived. Therefore, these results demonstrate that direct transfer technique of vitrified and one-step diluted bovine blastocysts can be applied easily and effectively with the higher pregnancy rate on field trial without the equipment and embryological skills.

• Current Research on Agriculture and Life Sciences
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• 제8권
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• pp.51-58
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• 1990

본(本) 연구는 micromanipultor를 이용하여 생쥐의 8세포기배와 상실배 그리고 배반포기배를 양분(兩分)후 생존성을 검토하고, 또한 배반포기배를 양분후, 선별(選別)및 배양 과정없이 암컷 생쥐에 이식(移植)하는 경우 새끼쥐 생산의 가능성을 검토하고자 수행된 것이다. 그 경과를 요약해 보면 다음과 같다. 1. 생쥐의 8세포기배(細胞期胚)와 상실배(桑實胚)를 양분(兩分)하여 M2에 배양한결과 각각 64%, 81%가 배반포기배(胚盤胞期胚)까지 발생(發生)하였다. 2. M2 배양액에서 발생시킨 배반포기배를 Ham's F-10에서 배양한 결과 8세포기배에서는 86%, 상실배에서는 90%가 정상적으로 outgrowth 되었다. 3. 배반포기배를 배양 과정없이 바로 양분(兩分)하여 Ham's F-10에서 배양한 결과 97%가 정상적으로 outgrowth 되었으나 암컷생쥐에 이식한 결과 산차는 얻지 못하였다.

• Journal of Plant Biotechnology
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• 제35권1호
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• pp.47-53
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• 2008

겨우살이의 캘러스 형성 및 종자 발아 와 haustorium 발달에 요구되는 환경요인 및 생장조절제의 영향을 조사하였다. 아울러 종자의 인위부착을 통한 식물체 재생 체계를 확립하였다. 겨우살이 조직관찰 결과 종자에는 1개 또는 2개의 막대모양의 접합자배가 존재하며 종자의 외부에 위치한 유근 부위에 주로 세포분열이 왕성한 세포층이 존재하며 반대로 자엽은 종자 내부에 위치하며 상대적으로 분열능을 가진 세포수가 감소하였다. 겨우살이 종자 발아 와 haustorium 발달에 요구되는 여러 요인 중에서 광의 요구도가 절대적이었다. 본 실험에 조사된 다른 배양 환경요인 및 생장조절제들은 광의 효과를 대체하지 못하였으며 ethylene의 경우 발아촉진 효과가 3배 이상 증가함을 알 수 있었다. 여러 조직 중에서 오로지 겨우살이 화아로부터 캘러스 형성이 가능하였으며 캘러스 형성빈도는 $0.1\;mgl^{-1}$ IAA가 첨가된 B5 배지에 배양 시 27.3%로 가장 높았다. 본 연구에서 확립된 겨우살이 캘러스 배양 및 종자 인위 부착을 통한 식물체 재생체계는 겨우살이의 유용물질 생산 연구 및 대량증식 연구분야에 활용이 가능할 것으로 기대된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권3호
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• pp.340-345
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• 2008

Oxidative stress is one of the major causes of failure of in vitro storage of boar semen. Reactive oxygen species (ROS) are one of the important mediators of oxidative stress during in vitro storage of boar semen. Our study examined the effects of taurine on sperm characteristic and on in vitro developmental embryos during in vitro storage of boar semen for 7 days. Semen was randomly aliquoted into 3 centrifuge tubes and treated with different concentrations of taurine (25-100 mM). The characteristics of boar sperm were analyzed for motility by light microscopy, viability by using a Makler counting chamber and membrane integrity by a hypoosmotic swelling test (HOST). The percentages of motile spermatozoa in taurine groups after 5 days were significantly higher compared to the control. Sperm viability in the control was lower than in taurine groups after 7 days irrespective of different taurine concentration. In the hyoosmotic swelling test (HOST), significantly higher results were obtained in taurine groups after 3 days. Also, the developmental rates of IVM/IVF porcine embryos from semen treated with pyruvate and taurine were significantly increased when compared with the control (p<0.05). These results indicate that supplementation of taurine as an antioxidant in boar semen extender can improve the semen quality.

• 대한바이러스학회지
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• 제26권1호
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• pp.1-8
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• 1996

Human cytomegalovirus (HCMV) replication and $Ca^{2+}$ response in human cell lines of neuronal origin were investigated. SK-N-SH (neuroblastoma cells) and A172 cells (glioblastoma cells) were used. SK-N-SH cells were permissive for HCMV multiplication with a delay of one day compared to virus multiplication in human embryo lung (HEL) cells. The delay of HCMV multiplication in SK-N-SH cells appeared to be correlated with a delay in the $Ca^{2+}$ response. The cytoplasmic free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) began to increase at 12 h p.i. in HCMV-infected SK-N-SH cells, while $[Ca^{2+}]_i$ increase in HCMV-infected HEL cells was observed as early as 3 h p.i. On the whole, the level of the increase in $[Ca^{2+}]_i$ in SK-N-SH cells was about 30% of that in HEL cells. On the other hand, in A172 cells infected with HCMV, neither production of infectious virus nor detectable increase in $[Ca^{2+}]_i$ was observed. Treatment with TPA of HCMV-infected SK-N-SH cells resulted in $[Ca^{2+}]_i$ increase at 6 h p.i. The stimulatory effect of TPA on HCMV- induced $[Ca^{2+}]_i$ increase continued until 12 h p.i., but TPA failed to stimulate the $Ca^{2+}$ response in SK-N-SH cells at 24 h p.i., suggesting that the effect of TPA had disappeared in SK-N-SH cells at that time point. In conclusion, SK-N-SH cells are permissive for HCMV replication and the delay in $Ca^{2+}$ response may be a consequence of the lower responsiveness of SK-N-SH cells than HEL cells to HCMV infection.

• Asian-Australasian Journal of Animal Sciences
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• 제22권4호
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• pp.483-491
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• 2009

In vitro produced porcine embryos have potential application in reproductive biotechnology. However, their development potential has been very low. This study evaluated the in vitro developmental ability and quality of cloned and parthenogenetic porcine embryos having 2-4 cells or 5-8 cells on Day 2 of in vitro culture. Analysis of results showed that 2 to 4 cell embryos had higher ability to form blastocysts than 5 to 8 cell embryos (p<0.05). Blastocysts produced from culture of 2 to 4 cell embryos also contained higher cell numbers and had lower BAX:BCLxL transcript ratio than those produced from 5 to 8 cell embryos (p<0.05), thereby suggesting 2 to 4 cell embryos have higher development potential. Further investigation revealed that 5 to 8 cell embryos had higher incidence (100${\pm}$0.0%) of blastomeric fragmentation than 2 to 4 cell embryos (15.2${\pm}$5.5% for parthenogenetic and 27.7${\pm}$7.1% for cloned embryos). This suggests that low development potential of 5 to 8 cell embryos was associated with blastomeric fragmentation. In conclusion, we have shown that morphological selection of embryos based on cell number on Day 2 of in vitro culture could offer a practical and valuable non-invasive means to select good quality porcine embryos.

• Asian-Australasian Journal of Animal Sciences
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• 제13권7호
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• pp.897-900
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• 2000

The experiment was conducted to study the effect of GnRH administration at induced estrus on pituitary and ovarian response in buffalo heifers. Eight Murrah river buffaloes of 12 to 13 months of age were treated with $PGF_{{2}{\alpha}}$ and eCG combination. GnRH (Fertagyl) 200 ug was injected (iv) at estrus in four heifers (treated group) while saline (2 ml, iv) was injected in remaining four heifers (control group). Blood was collected through jugular catheter to estimate plasma FSH, LH and estradiol level. The pretreatment plasma FSH, LH and estradiol values ranged from $8.46{\pm}1.97ng/ml$ to $12.31{\pm}1.30ng/ml$, $0.87{\pm}0.21ng/ml$ to $1.19{\pm}0.29ng/ml$ and $19.09{\pm}2.38pg/ml$ to $20.24{\pm}1.00pg/ml$ respectively. The plasma estradiol concentration elevated significantly (p<0.05) within 24 hr after eCG administration and reached its peak levels of $154.09{\pm}17.28pg/ml$ and $181.95{\pm}31.82pg/ml$ at estrus in respectively treatment and control groups. The plasma FSH and LH concentrations did not increase during follicular development after eCG administration while initial significant (p<0.05) increases in both plasma FSH and LH concentrations occured within 5 and 10 min, reaching peak levels of respectively $110.06{\pm}23.56ng/ml$ and $13.15{\pm}3.13ng/ml$ within 90 min after GnRH injection was detected. A sharp and significant decline in plasma estradiol concentration ($59.27{\pm}8.78pg/ml$) associated with synchronized ovulation within 24 hours after GnRH injection was recorded. The observation suggest that the hypophysis of prepubertal buffaloes treated with eCG have gonadotrophins awaiting the releasing factor to evoke release of gonadotrophin during the follicular phase to induce synchronized ovulation.