• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.369-375
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• 2011

The aim of this study was to evaluate the effect of ${\alpha}$-tocopherol on blastocysts development and subsequent cryosurvival of the vitrification. The ${\alpha}$-tocopherol(0, 100, 200, 400 ${\mu}M$) was added in to culture medium for the bovine embryos. The blasocysts from the ${\alpha}$-tocopherol and untreated control groups were then frozen-thawed, and their cryosurvival was assessed by in vitro culture for 48 h. There were no differences in the overall cleavage rate($56.14{\pm}4.66$, $58.18{\pm}4.70$, $62.97{\pm}6.86$ and $51.17{\pm}7.28$) among four treatment groups. However, in blastocyst development and total cell number were significantly higher in ${\alpha}$-tocopherol 200 ${\mu}M$ ($38.60{\pm}7.12$; $106.33{\pm}3.50$) to culture medium than other treatment groups($29.30{\pm}5.24$, $31.60{\pm}7.12$ and $26.37{\pm}4.18$; $101.36{\pm}5.12$, $97.27{\pm}2.87$, and $91.23{\pm}7.52$ respectively). Before and after vitrification, the total cell number and blastocyst development of embryo were significantly higher in July to August than September to October. In conclusion, addition of ${\alpha}$-tocopherol 200 ${\mu}M$ to in vitro bovine embryo culture medium was beneficial for improving embryo quality by decreasing the embryo damage blsstocysts cell number and improving the tolerance of the embryos to cryopreservation.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.157-162
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• 2006

It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs (cumulus oocytes complexes) in vitro. Attempts had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of FGF (fibroblast growth factor) on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in HPM 199 (Inst. of Functional peptide, Japan) containing 0.1, 1 and 10 ng/ml FGF for 24 hr, maturation rates to metaphase II ($70.0{\sim}75.0%$) were significantly higher (p<0.05) than that of control group (0 ng/ml FGF, 37.5%). When matured COCs with FGF were cultured in maturation medium after in vitro fertilization, developmental rates to blastocysts were 9.5, 0 and 2.9%, respectively, compared to 25.0% of the control group (p<0.05). When the matured COCs with FGF were cultured in HPM 199 (IFP971, Inst. of Functional peptide, Japan) containing 10% FBS, 0.8% BSA or 0.1% PVA (polyvinyl alcohol), the blastocyst formation rates were 12.4, 12.8 and 8.5%, respectively, while the rates of matured COCs with FGF and cultured with IVMD and IVD (Inst. of Functional peptide, Japan) without serum were 38.4% and 34.8%, respectively (p<0.05). These results suggested that FGF is available for in vitro maturation of bovine COCs and is not suitable for in vitro development, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine embryo development.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.277-282
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• 2011

The present research was carried out to evaluate the possibility of increasing female offspring production ratios using artificial insemination buffer (AIB) before artificial insemination (AI). In this experiment, we optimized AIB composition, made an AIB gun and analyze factors affecting AI non-return rate after AIB treatment. The AIB was made with the base of Tris-buffer supplemented with L-arginine and several other chemicals that might reduce the motility of male sperm compared to the female counterpart, therefore, increasing the possibility of fertilization by female sperm. AIB must be deposited into $2^{nd}$ to $4^{th}$ cervix by AIB gun. After 15 min of AIB deposition, frozen semen was deposited into the same place. A total of 348 cattle were inseminated with AIB insemination, and there were no significant differences between AIB and traditional AI non-return rates (56.8% vs. 55.7%). The AI non-return rate in AIB group, however, differed significantly among 7 Hanwoo farms. The parturition numbers ($1^{st}$ to $7^{th}$) of cows did not affect AIB AI rate. The proportion of AIB AI success rates was significantly higher in Hanwoo cows than in dairy cows (61.0% vs. 48.7%), but the average AI success rate did not differ significantly between AIB and conventional AI (56.8% vs. 55.7%). The female offspring production rate in $2^{nd}$ to $4^{th}$ cervix deposition place was significantly higher than that in the uterus body (77.7% vs. 59.6%, p<0.05). The injection volume of AIB in 5 and 10 ml was significantly higher than that in 2 ml (77.7%, 78.7% vs. 51.8%, p<0.05), but there were no differences in AIB injection volume between 5 and 10 ml. The best exposure time of AIB in the cervix was 10 to 15 min rather than 5 min (79.2%, 77.2% vs. 52.6%, p<0.05). AIB therefore needs to have an exposure time of at least over 10 min for a higher production rate of female offspring. In conclusion, AIB could be used in AI industry to increase the female offspring ratio and AIB AI can increase the AI success rate.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.101-105
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• 2007

본 연구는 한우와 홀스타인 젖소에서 연속적인 다배란 처리 시 효율성 및 부작용에 대한 문제점을 연구하고자 다른 종간 다배란 처리효과를 비교, 조사하였다. 연속적인 다배란 처리를 위하여 estradiol benzoate(EB)와 CIDR plus, FSH 호르몬을 이용하여 한우에서는 17두, 젖소에서는 9두를 2회 이상 반복 이용하였다. 다난포 발생을 위해서는 FSH 호르몬을 이용하고 호르몬 처리 동기화를 위해서는 CER plus 기구를 질 내 삽입하였으며, 주요 조사 항목으로는 번식기관의 문제점, 배란 효율, 회수된 수정란 수, 우수 수정란 수 등을 분류하고, 종합적으로 종간 연속적인 다배란 효과를 비교, 조사하였다. 한우에서는 17두 중 4두(23.5%)에서 자궁경관의 협착이 발생하였고, 자궁 내에 손상이 발생하여 채란이 되지 않는 경우도 4두(23.5%)의 발생이 조사되었으나, 상대적으로 젖소에서는 9두 중 4두에서 일시적으로 배란이 전혀 일어나지 않는 경우가 있었다. 연속 처리 후 채란된 수정란의 수를 종합해 보면, 젖소에서는 평균 $4.3{\pm}3.4$개, 한우에서는 $2.7{\pm}2.5$개였으며, 종간 호르몬 효율과 연관된 배란율 조사에서는 한우에서는 89.7%, 젖소에서는 80.4%의 성적을 보였다. 또한, 3회까지 연속 다배란 처리를 한 결과, 한우나 젖소에서 채란된 수정란의 수는 비슷한 수준으로 채란되었다. 이와 같은 성적을 종합해 볼 때 EB제나 CIDR plus, FSH를 이용하여 1개월 주기로 연속적인 수정란 채란을 위한 다배란 처리가 소의 종에 무관하게 가능하다고 사료되지만 일시적으로 배란이 일어나지 않는 소에 대한 문제나 번식기관의 손상, 채란되는 수정란의 수가 적다는 것에 더 많은 연구가 필요하다고 사료된다.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.163-169
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• 2014

The objective of this study was to evaluate the characteristics of Korean Native Cattle sperm frozen-thawed with L-cysteine and/or catalase. The semen from bulls was collected by the artificial vagina method, and Triladyl containing 20% egg-yolk and/or L-cysteine (L), catalase (C) and L-cysteine + catalase was added to the diluted semen for cryopreservation. The results showed that sperm viability was significantly higher in the L-cysteine + catalase ($69.49{\pm}3.16%$) group than in the control ($60.5{\pm}3.94%$) group (p<0.05). Acrosome damage was significantly lower in the L-cysteine ($17.12{\pm}1.08%$) group than in the control ($21.46{\pm}1.14%$), catalase ($20.54{\pm}0.76%$), and L-cysteine + catalase ($19.29{\pm}0.65%$) groups (p<0.05). In addition, the level of intact mitochondria in the spermatozoa was significantly higher in the L-cysteine ($58.65{\pm}1.39%$) group than in the control ($50.63{\pm}2.37%$) group (p<0.05). The hydrogen peroxide level in the frozen-thawed sperm was significantly lower in the L-cysteine ($3.74{\pm}1.66%$), catalase ($4.65{\pm}1.87%$), and L-cysteine + catalase ($8.11{\pm}2.15%$) groups than in the control ($13.22{\pm}1.6%$) group (p<0.05). The glutathione level was significantly higher in the L-cysteine ($1.33{\pm}0.03%$) group than in the control ($1.08{\pm}0.06%$), catalase ($1.05{\pm}0.02%$) and L-cysteine + catalase ($1.11{\pm}0.03%$) groups (p<0.05). In conclusion, L-cysteine and catalase could protect the membrane of Korean Native Cattle sperm from damage during sperm cryopreservation. Especially, L-cysteine was more effective for keeping acrosomes and mitochondria intactness during sperm cryopreservation.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.297-303
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• 2014

The objective of this study was to determine the breed differences in the 3'-untranslated region (UTR) of MC1R mRNA, which may be used to distinguish Korean brindle cattle (Chikso) from other breeds. We investigated the relationship between the variation of 3'-UTR of the MC1R mRNA and coat color among different breeds and the Korean brindle cattle with different coat colors. MC1R mRNA expression levels were determined in accordance with the coat color and hair colors of the tail. Total cellular RNA was extracted from the hair follicles of the tails in Hanwoo, Korean brindle cattle, Holstein and $Hanwoo{\times}Holstein$ crossbred cattle. After cDNA synthesis, PCR was performed. Sequences of the 3'-UTR of MC1R mRNA were analyzed. The 3'-UTR of the MC1R mRNA from different breeds of cattle did not show any variations. There were no variations in the 3'-UTR of the MC1R mRNA in Korean brindle cattle with different coat colors. The levels of MC1R mRNA expression in hair follicles of the tail varied substantially among the Korean brindle cattle with different coat colors, except yellow coat color. Correlation between the MC1R mRNA expression in the hair follicles of the tail and coat color may be present in the Korean brindle cattle, but not between the variations of 3'-UTR of MC1R mRNA and coat color. Further studies to determine the regulation of MC1R mRNA expression from the hair follicles of different coat colors will be beneficial in clarifying the role of MC1R in the coat colors of the Korean brindle cattle.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.177-182
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• 2006

This study was conducted to investigate the effects of melengesterol acetate (MGA) and $PGF_{2{\alpha}}$ administrations on serum progesterone level, synchrony of estrus and conception rates in Han-woo. Firstly, ten heifers and one freematin were fed 0.5 mg MGA/day for 14 days in a grain carrier, and after 19 days of MGA feeding, a single injection of 25 mg $PGF_{2{\alpha}}$ were treated. Blood samples were collected to evaluate serum progesterone concentrations from the start of feeding of MGA until the end of feeding and subsequent estrous detection and artificial insemination (AI) at 3 days intervals, and on days of $PGF_{2{\alpha}}$ injection, estrous detection, AI, and 15th and 60th days after AI. The level of progesterone in the blood began to increase from 7 days after MGA feeding, and 9 days after feeding it became 5.4 ng/ml and maintained that level thereafter. On the 33th day when the $PGF_{2{\alpha}}$ was injected, it reached the peak level of 7.6 ng/ml. However, 2-3 days after $PGF_{2{\alpha}}$ injection, it dropped to 1.4 ng/ml drastically (p<0.05). Secondly, one hundred and ninety four Hanwoo heifers or cows were divided into two groups to compare estrous induction and conception rates: the one treated with MGA and $PGF_{2{\alpha}}$, (n=104) and the other with $PGF_{2{\alpha}}$ treatment (two injections at 11 days interval, n=90). The heifers or cows treated with MGA and $PGF_{2{\alpha}}$ were identical to those used as above. The percentages of heifers or cows showed estrus were higher in the $MGA+PGF_{2{\alpha}}$ treatment (91.3%) than in the $PGF_{2{\alpha}}$ treatment (72.2%, p<0.05). Conception rates were also higher in the $MGA+PGF_{2{\alpha}}$ treatment (94.2%) than in the $PGF_{2{\alpha}}$ treatment (88.9%, p<0.05). The results of this experiment indicate that estrus synchronization using $MGA+PGF_{2{\alpha}}$ is more effective than that using $PGF_{2{\alpha}}$ (two injections) in Hanwoo.

• Journal of Embryo Transfer
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• v.21 no.4
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• pp.307-313
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• 2006

The present study was conducted to investigate the effects of time of vaccination in recipients on the pregnancy rate and the viability of calves derived from embryos produced in vitro. In experiment 1, control group was non-vaccinated, group 1-1 received vaccine (Pfizer, Exton, PA, USA) during 0$\sim$4 weeks and group 1-2 received vaccine during 4$\sim$8 weeks before embryo transfer. The pregnancy rates in the control (42.6%) and group 1-2 (45.3%) were significantly higher (p<0.05) than that in the group 1-1 (32.6%). However, the abortion rates were similar among groups (4.9 to 13.5%). In experiment 2, the recipients received embryos produced in vitro were non-vaccinated (control) or vaccinated. Vaccine was injected during 0$\sim$4 weeks (group 2-1) and 4$\sim$8 weeks (group 2-2) before parturition. The incidence of a disease in calves was significantly higher in the control (22.4%) than in other vaccinated groups (2.2% and 3.1%, p<0.05). The mortality of calves in the control is 27.6%, which was significantly higher than that of group 2-1 and group 2-2 (11.1% and 7.8%).

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.89-95
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• 2005

In this study, we investigated the effects of number, grade, stage and day of embryos on pregnancy and abortion rates of Holstein heifer recipients after transfer of in-vitro produced Korean Native Cattle (Hanwoo) embryos. When one, two or three blastocysts were transferred to recipients, pregnancy (32.0, 44.0 and $32.0\%$) and abortion (12.5, 13.6 and $6.3\%$) rates did not differ among treatments. The pregnancy and abortion rates of different grades of blastocysts were $40.8\~44.2\%\;and\;9.4\~16.3\%$, respectively. However, they were not significantly different. According to the stage of blastocyst, the pregnancy rates were similar between $39.1\%\;and\;45.3\%$. However, the abortion rate in hatched blastocyst stage $(20.8\%)$ was significantly higher than that in late blastocyst stage $(9.0\%)$. The pregnancy rates in day $7\cdot7$ and day $7\cdot8$ blastocyst (47.1 and $53.9\%$, respectively) was significantly higher than that in the day $9\cdot9\;and\;8\cdot9$ blastocyst (29.8 and $27.7\%$, respectively). The abortion rate was the highest in the day $9\cdot9$ blastocyst $(30.8\%)$ among treatments, and it was significantly different compared to that in the day $7\cdot7$ blastocyst $(10.1\%)$. Therefore, this study suggested that the pregnancy rate was affected by only day of embryos, while the abortion rate was affected by stage and day of embryos.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.167-172
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• 2007

Pituitary-specific transcription factor (PIT1) 유전자는 동물의 성장을 조절하고 근육 형성에 관여하는 유전자로서 최근에는 단일염기다형성 변이가 한우를 비롯한 동물에서 관찰되었으며, 한우의 경제 형질과 연관성이 보고되었다. 본 연구는 PIT1 유전자의 단일염기다형성 변이가 한우에서 성장 인자에 미치는 영향과 경제 형질에 대한 유전자형간 육종가와의 상관성에 대해 알아보고자 하였다. 도체 성적을 보유하고 있는 한우 후보종모우 집단 268두를 대상으로 PIT1 유전자 A1256G 다형성을 조사하여 유전자형의 빈도를 분석하였고 각각의 유전형에 따른 기본적인 검정 성적을 바탕으로 경제 형질과의 연관성을 비교 분석하였다. 268두의 한우에서 PIT1 유전자의 A1256G 유전자형 빈도는 MseI 제한 효소를 사용했을 때 A 유전자 빈도(0.37)보다 G 유전자 빈도(0.62)가 높게 나타났다. 통계적 분석을 통하여 각 유전자형에 대한 경제 형질과의 관련성을 분석한 결과, 각 유전자형 간에 12개월령 체중 (body weight 12, BW12)에서 유의한 차이를 보였고, 등지방 두께 육종가 (Backfat thickness-estimated breeding value, BF-EBV)와도 유의한 차이가 있었지만 (p<0.05), marbling score (MS), carcass weight (CW), M. longissimus dorsi area (LDA) 등 다른 경제 형질과는 통계학적으로 유의한 차이가 없었다. PIT1 유전자의 A1256G 다형성은 한우의 성장과 도체체중에 관여하는 인자로 작용하는 것으로 보여진다.